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Topical tretinoin (retinoic acid) treatment of hyperpigmented lesions associated with photoaging in Chinese and Japanese patients: a vehicle-controlled trial.

BACKGROUND: Hyperpigmented lesions are a predominant component of photoaging in Chinese and Japanese persons. Topical 0.1% tretinoin cream improves the hyperpigmentation associated with photoaging in Caucasian persons. OBJECTIVE: Our purpose was to assess the efficacy of 0.1% tretinoin cream treatment of hyperpigmented lesions associated with photoaging in Chinese and Japanese patients. METHODS: Forty-five photoaged patients (23 Chinese, 22 Japanese) completed a double-blind, randomized study in which 21 applied 0.1% tretinoin cream and 24 applied vehicle cream once daily to face and/or hands for 40 weeks. Patients' hyperpigmented lesions were evaluated clinically and by colorimetry throughout the study and by histologic analysis of skin biopsy specimens taken before therapy and at the end of treatment. RESULTS: At the end of treatment, hyperpigmented lesions of the face and hands were lighter or much lighter in 90% of patients receiving tretinoin compared with 33% receiving vehicle (p < 0.0001). Colorimetry demonstrated significant lightening of lesions after tretinoin compared with vehicle (p < 0.05). Histologic analysis of hyperpigmented lesions demonstrated a statistically significant 41% decrease in epidermal pigmentation with tretinoin therapy as compared with a 37% increase in the vehicle group (p = 0.0004). No patient withdrew for adverse effects. CONCLUSION: By clinical, colorimetric, and histologic evaluation, 0.1% tretinoin cream significantly lightens the hyperpigmentation of photoaging in Chinese and Japanese patients.

Administration, Topical↗

Combined effects of irritants and allergens. Synergistic effects of nickel and sodium lauryl sulfate in nickel- sensitized individuals.

Knowledge of the combined effects of irritants and allergens is of interest with respect to accurate risk assessment. The threshold for elicitation of allergic contact dermatitis in previously sensitized individuals may theoretically be markedly influenced by the simultaneous presence of irritants and allergens. Combined exposures have, however, only been studied infrequently. In the present study, the combined effect of an irritant and an allergen was evaluated in a dose-response designed experimental study. 20 nickel-sensitized subjects were exposed to patch testing with varying concentrations of NiCl2 (nickel chloride) and sodium lauryl sulfate (SLS) alone and in combination. Evaluation of skin reactions was performed by colorimetry, measurement of transepidermal water loss and clinical evaluation, and the data were analyzed by logistic dose-response models. A synergistic effect was found of combined exposure to NiCl2 and SLS, as compared to each of the substances applied separately, as evaluated by colorimetry and clinical scoring. This means that the effect produced by the combined exposure was substantially greater than the effect produced by either of the substances alone. A synergistic effect of combined exposure on skin barrier impairment was not found, since the barrier function is significantly influenced by SLS-exposure only and not by NiCl2. Concentration limits are used by industry and government agencies to protect consumers. The present results clearly illustrate that elicitation thresholds and concentration limits may be influenced considerably by combined exposure to allergens and irritants.

Adult↗

Topical tretinoin (retinoic acid) therapy for hyperpigmented lesions caused by inflammation of the skin in black patients.

BACKGROUND AND METHODS: Irregular disfiguring skin hyperpigmentation due to inflammation may develop in black persons. We investigated the treatment of this hyperpigmentation with topical tretinoin (0.1 percent retinoic acid cream). Fifty-four subjects completed a 40-week randomized, double-blind, vehicle-controlled study. Twenty-four subjects applied tretinoin daily to the face, arms, or both areas, and 30 subjects applied vehicle cream. At base line and after 40 weeks of treatment, each subject's post-inflammatory hyperpigmented lesions and normal skin were assessed by clinical and colorimetric evaluations and by analysis of biopsy specimens. RESULTS: The facial post-inflammatory hyperpigmented lesions of the tretinoin-treated subjects were significantly lighter after the 40 weeks of therapy than those of the vehicle-treated subjects (P < 0.001); overall improvement was first noted after four weeks of tretinoin treatment. At the end of treatment, colorimetry demonstrated a 40 percent lightening of the lesions toward normal skin color in the tretinoin-treated lesions, as compared with an 18 percent lightening in vehicle-treated lesions (P = 0.05). The epidermal melanin content in the lesions decreased by 23 percent with tretinoin and by 3 percent with vehicle (P = 0.24). Normal skin was minimally lightened by tretinoin as compared with vehicle, according to both clinical evaluation (0.1 vs. -0.1 unit change on an 8-point scale; P = 0.055) and colorimetry (P < 0.001). Retinoid dermatitis developed in 12 of the 24 tretinoin-treated subjects who completed the study (50 percent) and in 1 tretinoin-treated subject who withdrew from the study, but diminished as the study progressed. CONCLUSIONS: Topical application of tretinoin significantly lightens post-inflammatory hyperpigmentation and, to a clinically minimal but statistically significant degree, lightens normal skin in black persons.

Administration, Cutaneous↗

Precision of laser Doppler scanning in clinical use.

Reactions of the cutaneous microcirculation to local stimuli have been quantitated in terms of area of erythema (planimetry), change in colour (colorimetry) and time-dependent changes in perfusion (one-dimensional laser Doppler fluxmetry (LDF)). Using the new technique of laser Doppler scanning (LDS) it is now possible to quantify and demonstrate two-dimensional patterns of microvascular perfusion in terms of intensity and extent. The precision of this new technique was compared with the previously established techniques by examining the reactions of 15 healthy volunteers to the intracutaneous injection of carbacholin and isotonic NaCl solutions. Both stimuli were applied paravertebrally, on the left and right side. Reactions were examined after 10 min. As a control for temporal reproducibility the examination was repeated on another day. All methods allowed the differentiation of reactions to carbacholin from those to NaCl (P < or = 0.0001). Only colorimetry and LDS provided data that were unequivocally reproducible when comparing both sides (r > or = 0.78). Using LDF or planimetry, reproducibility of values was lower (r < or = 0.58). When looking at measurements performed on different days, only values of LDS were reproducible (r > or = 0.77). Hyperaemic areas seen with LDS were significantly larger than those determined by planimetry. Results of LDS are reproducible spatially and temporally: the technique will allow time-course observations of skin reactions that are accompanied by medium-term changes in perfusion.

Adult↗

Spectral Transforms as a Tool to Optimize Digital Phenotyping in Biological Images.

Modern livestock breeding has mastered genotyping. Genome-wide association studies, genomic selection, and SNP arrays enable genetic merit prediction at lower cost. However, phenotyping remains the bottleneck, as manual measurement is slow, expensive, subjective, and unable to capture spatial or temporal trait organization. Digital phenotyping via artificial intelligence could resolve this, but deep learning requires thousands of labelled examples, impractical when phenotyping cost itself limits datasets to hundreds of individuals. This creates a paradox: AI could accelerate phenotyping but requires large numbers of samples to train the models. Here, we demonstrate that integrating computer vision with machine learning offers sample-efficient digital phenotyping using eggshell colour as a model system. Rather than learning features from scratch (deep learning), we engineer physically motivated features via Wavelet transforms that decompose images into multi-scale spatial components. Wavelet features captured 14.2 percentage points more variance (R2&#x2009;=&#x2009;0.976 vs. 0.834, p&#x2009;<&#x2009;0.001) than standard colorimetry, with 50% better sample efficiency (achieving at n&#x2009;=&#x2009;60 what colorimetry required n&#x2009;=&#x2009;120). Variance decomposition revealed 77% of discriminative capacity derives from spatial patterns (bands, spots, gradients) invisible to scalar averages. Additionally, we identified "cryptic phenotypes" (3.3%) where spatial patterns contradicted average colour, cases where colorimeters failed but Wavelets succeeded. The underlying principle-that spatial decomposition can recover organizational information lost by scalar averaging-may be applicable to other traits with spatial or temporal structure, such as marbling, dermatitis, or pigmentation rhythms, although whether comparable performance gains would be observed remains to be tested empirically. Hence, for breeding programs implementing genomic selection, computer vision-based digital phenotyping captures complex trait variation without massive training datasets, addressing the bottleneck that increasingly limits genetic progress as genotyping becomes trivial.

Wavelet transform↗

BAK overexpression mediates p53-independent apoptosis inducing effects on human gastric cancer cells.

BACKGROUND: BAK (Bcl-2 homologous antagonist/killer) is a novel pro-apoptotic gene of the Bcl-2 family. It has been reported that gastric tumors have reduced BAK levels when compared with the normal mucosa. Moreover, mutations of the BAK gene have been identified in human gastrointestinal cancers, suggesting that a perturbation of BAK-mediated apoptosis may contribute to the pathogenesis of gastric cancer. In this study, we explored the therapeutic effects of gene transfer mediated elevations in BAK expression on human gastric cancer cells in vitro. METHODS: Eukaryotic expression vector for the BAK gene was constructed and transferred into gastric cancer cell lines, MKN-45 (wild-type p53) and MKN-28 (mutant-type p53). RT-PCR and Western Blotting detected cellular BAK gene expression. Cell growth activities were detected by MTT colorimetry and flow cytometry, while apoptosis was assayed by electronic microscopy and TUNEL. Western Blotting and colorimetry investigated cellular caspase-3 activities. RESULTS: BAK gene transfer could result in significant BAK overexpression, decreased in vitro growth, cell cycle G0/G1 arrest, and induced apoptosis in gastric cancer cells. In transferred cells, inactive caspase-3 precursor was cleaved into the active subunits p20 and p17, during BAK overexpression-induced apoptosis. In addition, this process occurred equally well in p53 wild-type (MKN-45), or in p53 mutant-type (MKN-28) gastric cancer cells. CONCLUSIONS: The data presented suggests that overexpression of the BAK gene can lead to apoptosis of gastric cancer cells in vitro, which does not appear to be dependent on p53 status. The action mechanism of BAK mediated apoptosis correlates with activation of caspase-3. This could be served as a potential strategy for further development of gastric cancer therapies.

Apoptosis↗

Onchocerca parasites and Wolbachia endosymbionts: evaluation of a spectrum of antibiotic types for activity against Onchocerca gutturosa in vitro.

BACKGROUND: The filarial parasites of major importance in humans contain the symbiotic bacterium Wolbachia and recent studies have shown that targeting of these bacteria with antibiotics results in a reduction in worm viability, development, embryogenesis, and survival. Doxycycline has been effective in human trials, but there is a need to develop drugs that can be given for shorter periods and to pregnant women and children. The World Health Organisation-approved assay to screen for anti-filarial activity in vitro uses male Onchocerca gutturosa, with effects being determined by worm motility and viability as measured by reduction of MTT to MTT formazan. Here we have used this system to screen antibiotics for anti-filarial activity. In addition we have determined the contribution of Wolbachia depletion to the MTT reduction assay. METHODS: Adult male O. gutturosa were cultured on a monkey kidney cell (LLCMK 2) feeder layer in 24-well plates with antibiotics and antibiotic combinations (6 to 10 worms per group). The macrofilaricide CGP 6140 (Amocarzine) was used as a positive control. Worm viability was assessed by two methods, (i) motility levels and (ii) MTT/formazan colorimetry. Worm motility was scored on a scale of 0 (immotile) to 10 (maximum) every 5 days up to 40 days. On day 40 worm viability was evaluated by MTT/formazan colorimetry, and results were expressed as a mean percentage reduction compared with untreated control values at day 40. To determine the contribution of Wolbachia to the MTT assay, the MTT formazan formation of an insect cell-line (C6/36) with or without insect Wolbachia infection and treated or untreated with tetracycline was compared. RESULTS: Antibiotics with known anti-Wolbachia activity were efficacious in this system. Rifampicin (5 x 10(-5) M) was the most effective anti-mycobacterial agent; clofazimine (1.25 x 10(-5) M and 3.13 x 10(-6) M) produced a gradual reduction in motility and by 40 days had reduced worm viability. The other anti-mycobacterial drugs tested had limited or no activity. Doxycycline (5 x 10(-5) M) was filaricidal, but minocycline was more effective and at a lower concentration (5 x 10(-5) M and 1.25 x 10(-5) M). Inactive compounds included erythromycin, oxytetracycline, trimethoprim and sulphamethoxazole. The MTT assay on the insect cell-line showed that Wolbachia made a significant contribution to the metabolic activity within the cells, which could be reduced when they were exposed to tetracycline. CONCLUSION: The O. gutturosa adult male screen for anti-filarial drug activity is also valid for the screening of antibiotics for anti-Wolbachia activity. In agreement with previous findings, rifampicin and doxycycline were effective; however, the most active antibiotic was minocycline. Wolbachia contributed to the formation of MTT formazan in the MTT assay of viability and is therefore not exclusively a measure of worm viability and indicates that Wolbachia contributes directly to the metabolic activity of the nematode.

Journal Article↗

Analyses of cystine in human hair: its level in women's hair of former times.

An appropriate procedure for analysis of cystine in the hair has been established, and this procedure was applied to the hair of Japanese women. The recovery of authentic cystine added to a hair sample was 85-90% in 6N HCl hydrolysis, and 96% in acid hydrolysis after performic acid oxidation. The 18-hr acid hydrolysis, but not 4-hr one, was sufficient to digest the hair. In colorimetry by using phosphotungstate, Brown's reagent gave a stable color development. The cystine content by amino acid autoanalysis was significantly correlated with that by the phosphotungstate colorimetry by the modified method of Kassel et al. or of Shinohara. The cystine content in women's hair, which was collected from specimens of different individuals cut during the period from 1910s to 1980, indicated a wide variation ranging from 0.654 to 1.607 mmol half-cystine per g of hair after washing with 0.5% sodium laurylsulfate.

Adult↗

Stable transfection of extrinsic Smac gene enhances apoptosis-inducing effects of chemotherapeutic drugs on gastric cancer cells.

AIM: To explore the feasibility of enhancing apoptosis-inducing effects of chemotherapeutic drugs on human gastric cancer cells by stable transfection of extrinsic Smac gene. METHODS: After Smac gene was transferred into gastric cancer cell line MKN-45, subclone cells were obtained by persistent G418 selection. Cellular Smac gene expression was determined by RT-PCR and Western blotting. After treatment with mitomycin (MMC) as an apoptotic inducer, in vitro cell growth activities were investigated by trypan blue-staining method and MTT colorimetry. Cell apoptosis and its rates were determined by electronic microscopy, annexin V-FITC and propidium iodide staining flow cytometry. Cellular caspase-3 protein expression and its activities were assayed by Western blotting and colorimetry. RESULTS: When compared with MKN-45 cells, the selected subclone cell line MKN-45/Smac had significantly higher Smac mRNA (3.12+/-0.21 vs 0.82+/-0.14, t = 7.52, P<0.01) and protein levels (4.02+/-0.24 vs 0.98+/-0.11, t = 8.32, P<0.01). After treatment with 10 microg/mL MMC for 6-24 h, growth inhibition rate of MKN-45/Smac (15.8+/-1.2-54.8+/-2.9%) was significantly higher than that of MKN-45 (5.8+/-0.4- 24.0+/-1.5%, t = 6.42, P<0.01). Partial MKN-45/Smac cancer cells presented characteristic morphological changes of apoptosis under the electronic microscope with an apoptosis rate of 36.4+/-2.1%, which was significantly higher than that of MKN-45 (15.2+/-0.8%, t = 9.25, P<0.01). Compared with MKN-45, caspase-3 expression levels in MKN-45/Smac were improved significantly (3.39+/-0.42 vs 0.96+/-0.14, t = 8.63, P<0.01), while its activities were 3.25 times as many as those of MKN-45 (0.364+/-0.010 vs 0.112+/-0.007, t = 6.34, P<0.01). CONCLUSION: Stable transfection of extrinsic Smac gene and its over-expression in gastric cancer cell line can significantly enhance cellular caspase-3 expression and activities, ameliorate apoptosis-inducing effects of mitomycin C on cancer cells, which is a novel strategy to improve chemotherapeutic effects on gastric cancer.

Antibiotics, Antineoplastic↗

[The experimental study of the anti-enterovirus effects of drugs in vitro].

OBJECTIVE: To screen the safe and effective anti-enterovirus drugs for clinical application. METHODS: The cytotoxicity of Ribavirin, Shuanghuanglian and Garlic were evaluated through MTT colorimetry and cell morphology. The antiviral activity of Ribavirin, Shuanghuanglian and Garlic were studied in HEL and Vero cells infected with CBV3 and ECHO11 by observing cytopathic effect (CPE), MTT colorimetry and plaque-reduction assay. The antiviral activity of these three drugs were compared and that of Shuanghuanglian and Garlic were also compared before and after ECHO11 absorbing by plaque-reduction assay. RESULTS: (1) The cytotoxicity of these three drugs were expressed as TC50(50% toxic concentration). TC50 of Ribavirin was 2 mg/ml, of Shuanghuanglian 5 mg/ml, of Garlic 12.5 micrograms/ml. (2) Ribavirin could inhibit CBV3 and ECHO11 at the concentration ranged from 1 mg/ml to 1.5 mg/ml. Shuanghuanglian could inhibit CBV3 and ECHO11 at the concentration of 0.5 mg/ml and the viral inhibiting effect was concentration-dependent. Garlic could inhibit CBV3 and ECHO11 at the concentration ranged from 2.5 micrograms/ml to 7.5 micrograms/ml and 5 micrograms/ml was the most effective. (3) Plaque-reduction assay was used to test the anti-virus (CBV3 and ECHO11) activity of these three drugs: plaque reduction rate of 1.5 mg/ml Ribavirin was 43.2% and 37.2%, of 2.5 mg/ml Shuanghuanglian was 81.1% and 88.4% and of 5 micrograms/ml Garlic was 66.2% and 77.4% respectively. The plaque reduction rate of Ribavirin was lower than the other two drugs (P < 0.05), between these two showed no significant difference (P > 0.05). The anti-ECHO11 activity of Shanghuanglian before ECHO11 adsorbing was higher than after ECHO11 adsorbing (P < 0.05). There was no significant difference between the plaque reduction rates of Garlic before and after ECHO11 adsorbin. CONCLUSION: All of the three drugs have anti-virus activity in vitro while Shuanghuanglian and Garlic are more effective. Among these three drugs, the cytotoxicity of Shuanghuanglian is the most weak and the anti-virus activity is the strongest. The antiviral activity of Shuanghuanglian adding before virus adsorbing was higher, so it seems that Shuanghuanglian can prevent EV infection.

Animals↗

[Determination on chemical constituents of Curcuma L. produced in China].

OBJECTIVE: To provide some valuable information for the research on species systematization and quality standardization of Curcuma drugs. METHODS: Colorimetry, TLC-colorimetry and water vapour distillation were used to determine the contents of curcuminoids, curcumin and essential oils of 34 samples of Curcuma produced in China. RESULTS AND CONCLUSION: The contents of curcuminoids and curcumin are different upto an extent of over 100 times among the varieties of Curcuma, and those of essential oils to an extent of 6 times. But there is little difference in the samples of the same species from different producing areas. There is also an obvious correlativity between the contents of curcuminoids and essential oils in the varieties of curcuma.

Curcuma↗

[Effects of homoharringtonine on proliferation of human retro-ocular fibroblasts].

This is a study amied at the effects of homoharringtonine on the proliferation of human retro-ocular fibroblasts. The human retroocular fibroblasts (HROFb) were cultured and MTT colorimetry assay was used to determine the proliferation of HROFb. The influence of homoharringtonine on the proliferation of HROFb was observed in vitro. The results showed that MTT colorimetry assay could precisely reflect the proliferation of HROFb. The absorbance was directly proportional to the number of the cells (r = 0.996, P < 0.001). The proliferation of HROFb was inhibited by the homoharringtonine and the cell proliferation percentage was correlated positively to the homoharringtonine level (r = 0.6932, P < 0.001).

Cell Division↗

[Factors influencing the activity of fibrinolytic enzymes from earthworm, Eeisenia foetida].

OBJECTIVE: To study the factors influencing the activity of fibrinolytic enzymes from earthworm and to obtain the better way to extract fibrinolytic enzymes as well as keep its optimum activity. METHOD: 75% alcohol, 0.9% NaCl and 10% saccharose was used to extract the crude fibrinolytic enzymes from earthworm, the method of urokinase gelose-fibrin plate was used to measure the activity of fibrinolytic enzymes from earthworm. and the method of 3,3'-diaminobezidine tetrahydrochloride colorimetry to was used measure the content of selenium. The method use ts of measuring the content of arsenic was silver diethyldithiocarbamate colorimetry. RESULT: The fibrinolytin of earthworms reared with cattle soils had higher activity than that reared with garbage. The arsenic in the earthworm's body could improve the activity of earthworm's fibrinolytin. However, the selenium had litter influence on it. Among the three methods of extraction, the 75% alcohol one was the most efficient, the 0.9% NaCl was next, and the 10% saccharose was the lowest. The influence of dialysis on the activity of fibrinolytin was less than that of ultrafiltration, when the earthworm's fibrinolytin enzyme was further sublimated. CONCLUSION: The activity of the earthworm's fibrinolytin will be increased earthworm is reared with the fitting baits and when appropriate methods, of extraction and purification are used.

Animal Feed↗

[The effect of insulin on chemotherapeutic drug sensitivity in human esophageal and lung cancer cells].

OBJECTIVE: To discuss the effect of insulin, as a metabolic promoter, on chemotherapeutic drug sensitivity in human esophageal and lung cancer cells. METHODS: Human esophageal cancer cells NEC and human lung adenocarcinoma cells GLC were cultured and then inoculated into the wells. MTT was added. Chemotherapeutic drugs, etopside, cisplatin, or 5-fluoto-uracilum was added to examine their cytotoxity activity. Then insulin of the final concentration of 5 mU/ml was added 8 hours before etopside (30 - 40 micro g/ml), cisplatin (2.5 micro g/ml), or 5-Fu (50 micro g/ml) was added. MTT A value was tested by colorimetry to evaluate the number of cancer cells, cell activity, and metabolism status so as to reflect the cytotoxity of the anti-tumor agent. Insulin was added into the suspension of cancer cells. Flow cytometry was used to detect the cell-cycle progresses. RESULTS: Insulin alone did not inhibit the cell growth and mildly promoted the cell metabolism with the concentration > 5 mU/ml. Insulin (2.0 - 15.0 mU/ml) enhanced the chemocytotoxity of etopside (30 micro g/ml) on human esophageal and lung cancer cells as indicated by MTT colorimetry. GLC cell cycle assay showed that the S phase block induced by etopside, cisplatin and 5-FU and the G(2)/M block induced by 5-FU were enhanced by insulin with the increased block rates of 80% and 90% respectively. The increased block rate induced by insulin in NEC cells was lower than in GLC cells. CONCLUSION: A reversible metabolic promoter, insulin enhances the cytotoxity of the chemotherapeutic agents. It is possible to increase the growth and metabolism of cancer cells first so as to enhance the chemosensibility, and then administer chemotherapeutic agents, thus improving their therapeutic effects.

Adenocarcinoma↗

[Inhibitive effect of curcumin and amiloride on the fibrosis of rat hepatic stellate cells induced by oxidative stress].

OBJECTIVE: To investigate the effects of antioxidant curcumin and Na+/H+ exchanger inhibitor amiloride on the fibrosis of rat hepatic stellate cells (HSC) induced by oxidative stress in vitro. METHODS: HSC were incubated with 0.1 mmol/L ferric nitrilotriacetate complex (FeNTA). MTT colorimetry was used for assaying proliferation of HSC. Cytotoxicity was measured by LDH colorimetry. Collagen type I accummulation in the culture media was measured by ELISA. Intracellular malonildialdehyde (MDA), SOD, GSH and GSH-Px levels in the culture media were measured by respective reagent boxes. RESULTS: HSC incubation with FeNTA resulted in a significant production of intracellular MDA and GSH, associated with decrease of SOD and GSH-PX activity. Exposure of HSC to FeNTA significantly enhanced the number of proliferating HSC and collagen type I levels in the culture medium. All these effects were reversed by the antioxidant curcumin and by the Na+/H+ exchanger inhibitor amiloride. Combining curcumin with amiloride had better effects on the fibrosis of rat hepatic stellate cells induced by oxidative stress than curcumin or amiloride. CONCLUSION: The effects of combination of antifibrosis drugs curcumin and amiloride on different targets of HSC are superior to curcumin or amiloride.

Amiloride↗

[The effects of culture supernatant of human GRC-1 cells transfected with PF4 cDNA on the VEGF expression and the growth of ECV304 cells].

AIM: To construct the eukaryotic expression vector pcDNA3-PF4-SS, and to detect the effects of the culture supernatant of transfected GRC-1 cells on the VEGF expression in transfected GRC-1 cells and the growth of ECV304 cells. METHODS: The eukaryotic expression vector pcDNA3-PF4-SS was constructed and identified with Bgl II/BamH I digestion. The pcDNA3-PF4-SS was transfected stably into GRC-1 cells with lipofectamine mediation. The VEGF expression in transfected GRC-1 cells was detected by immunohistochemical staining, and the effect of the culture supernatant of transfected GRC-1 cells on ECV304 cells was detected by MTT colorimetry. RESULTS: Restrictive enzyme (Bgl II/BamH I)digestion analysis showed that the recombinant expression vector pcDNA3-PF4-SS had been constructed successfully. RT-PCR detection proved that hPF4 cDNA had been transfected into GRC-1 cells. The result of immunohistochemical staining showed that the VEGF expression could be seen in the cytoplasm and on cytomembrane of GRC-1 cells transfected with pcDNA3-PF4-SS, but the expression obviously weakened as comparison with that before transfection. Cell counting and MTT colorimetry manifested that the culture supernatant of transfected GRC-1 cells could inhibit markedly growth of ECV304 cells. CONCLUSION: The eukaryotic expression vector pcDNA3-PF4-SS has been constructed successfully, and stably transfected into the GRC-1 cells. The culture supernatant of transfected GRC-1 cells has obviously inhibitory effect on the growth of ECV304 cells and the VEGF expression in the GRC-1 cells, which lays some foundation for exploring the mechanism for anti-tumor growth and developing tumor vaccine for kidney neoplasms.

Carcinoma, Renal Cell↗

[Effect of IL-15 on the proliferation and differentiation of CD34+ cells from MDS patients].

AIM: To explore the effect of IL-15 on proliferation and differentiation of CD34 (+) cells from MDS patients. METHODS: The CD34 (+) cells were separated by magnetic bead-activated cell sorter (MACS) system, and cultured in fluid or methylcellulose semisolid medium. MTT colorimetry was used to examine the inhibitory effect of IL-15 on the proliferation of MDS CD34(+) cells.The numbers of CD34(+)cells and colony formation of CFU-E, BFU-E, CFU-G, CFU-GEMM were counted. The expressions of CD33, CD13, CD71, CD19 and CD3 on cultured cells and the change of cell-cycle were analyzed by FCM. RESULTS: The recovery rate of CD34(+) cells was (75.4+/-5.2) %, the purity of CD34 (+) cells in positive fraction was (90.3+/-6.3) % and the enriched rate was (83.1+/-12.5) % in 11 MDS patients. MTT colorimetry detection showed that IL-15 could promote the proliferation of MDS CD34(+) cells. The optimal time of promotoing CD34(+) cell proliferation by IL-15 was 8 days and optimal dosage of IL-15 was 20 microg/L. After culture for 8 days with 0 microg/L IL-15 (control group) and 20 microg/L IL-15(experimental group), the number of CD34 (+) cells increased by 4.6-fold in control group and 6.3-fold in experimental group (P<0.05, n=5). The colony formation rates of experimental group were significantly higher than those of control group. The expression rates of the CD molecules (except for CD3) on CD34(+) in experimental group were notably higher than those in control group. As compared with control group, much more CD34(+) cells of experimental group were in G(2) and S phase of cell cycle(P<0.05, n=7). CONCLUSION: IL-15 can obviously promote the proliferation and differentiation of CD34(+) cells from MDS patients.

Adolescent↗

A microsponge formulation of hydroquinone 4% and retinol 0.15% in the treatment of melasma and postinflammatory hyperpigmentation.

Disorders of hyperpigmentation such as melasma and postinflammatory hyperpigmentation (PIH) are common, particularly among people with darker skin types. Hydroquinone (HQ) bleaching creams are considered the gold standard for treating hyperpigmentation. Recently, a new formulation of HQ 4% with retinol 0.15% entrapped in microsponge reservoirs was developed for the treatment of melasma and PIH. Microsponges were used to release HQ gradually to prolong exposure to treatment and to minimize skin irritation. The safety and efficacy of this product were evaluated in a 12-week open-label study. A total of 28 patients were enrolled, and 25 completed the study. Study end points included disease severity, pigmentation intensity, lesion area, and colorimetry assessments. Adverse events also were recorded. Patients applied the microentrapped HQ 4% formulation to the full face twice daily (morning and evening). A broad-spectrum sunscreen was applied once in the morning, 15 minutes after application of the test product. Patients were evaluated at baseline and at 4, 8, and 12 weeks. The microentrapped HQ 4%/retinol 0.15% formulation produced improvement at all study end points. Improvement in disease severity and pigmentation intensity was statistically significant at weeks 4, 8, and 12 compared with baseline (P<.001). Lesion area and colorimetry measurements also were significantly improved at each visit (P<.001). Microentrapped HQ 4% was well tolerated, with only one patient discontinuing because of an allergic reaction, which was not considered serious. In this open-label study, microentrapped HQ 4% with retinol 0.15% was safe and effective.

Administration, Topical↗