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Biomedical subjects

Jie Luan

Publications and source records attributed to Jie Luan.

At least 19 recordsLinked to original sources

Mammalian cell entry protein of Mycobacterium tuberculosis induces the proinflammatory response in RAW 264.7 murine macrophage-like cells.

Mammalian cell entry (Mce1A) protein of Mycobacterium tuberculosis (Mtb) is involved in bacterial entry and survival in macrophages, which has been shown to induce production of tumor necrosis factor alpha (TNF-alpha). It remains unclear whether and how Mce1A functions upon the type I interleukin-1 receptor-associated protein kinase (IRAK-1) and TNF receptor-associated factor 6 (TRAF-6) of important proinflammatory cytokines. In this study, His-tagged Mce1A was expressed and purified. Also, two pieces of small interfering RNA (siRNA) were designed and synthesized by in vitro transcription, which exhibit specific and efficient silencing effect on mce1a expression. Furthermore, RAW 264.7 murine macrophage-like cells were exposed to His-tagged Mce1A or co-transfected with the Mce1A-expressing plasmid and efficient siRNA, and levels of IRAK-1 and TRAF-6 were then determined by Western blot. We show here that Mce1A induces up-regulations of IRAK-1 and TRAF-6 in macrophages in a dose-dependent manner. The level of Caspase-3 closely related with apoptosis was also determined, whereas no changes were observed. These results indicate that Mtb Mce1A protein induces a proinflammatory response in macrophages.

Animals↗

RNA interference effectively inhibits mRNA accumulation and protein expression of hepatitis C virus core and E2 genes in human cells.

RNA interference (RNAi) has been widely used for the analysis of gene function and represents a new promising approach to develop effective antiviral drugs. In this study, several small interfering RNAs (siRNAs) corresponding to two structural genes (core and E2) of hepatitis C virus (HCV) were designed and in vitro transcribed to explore the possibility of silencing these two genes. The plasmids pEGFP-C and pEGFP-E2, which contain the EGFP reporter gene and the core or E2 gene as silencing targets, were co-transfected with siRNAs into HEK 293T cells. At various time points of post-transfection, core and E2 expression levels were detected by fluorescence microscopy, flow cytometry, Western blotting, and real-time quantitative PCR. The results showed that the mean fluorescence intensity, protein expression, and RNA transcripts of siRNAs transfected cells were significantly reduced. This may provide an approach for the development of novel prophylactic or therapeutic agents for HCV infection.

Blotting, Western↗

[Breast reconstruction with a combined skin flap of DIEP and TRAM].

OBJECTIVE: To evaluate breast reconstruction with a combined skin flap of the deep inferior epigastric perforator (DIEP) and the transverse rectus abdominis musculocutaneous (TRAM). METHODS: The DIEP and TRAM united flap was elevated with the vessel pedicle of the deep inferior epigastric perforator on the affected side and the rectus abdominis muscle pedicle on the intact side. The reconstructive breast was shaped after the deep inferior epigastric vessels were anastomosed to the internal mammary vessels or the thoracodorsal vessels ipsilaterally. RESULTS: We have used the DIEP and TRAM united flaps for breast reconstruction in 17 cases. All of the flaps survived, and the reconstructed breasts were well-shaped with the follow-up of 6-18 months. CONCLUSIONS: The DIEP and TRAM united flap possesses of advantages such as rich blood supply, abundant tissue volume and easy shaping. It is especially applicable to the cases who have large chest defect and need large volume tissue.

Adult↗

[Lower capsular contracture of miniprostheses as compared with the conventional silicone implant].

OBJECTIVE: To determine if miniprostheses would form a capsule of significantly different biophysical, biochemical and histologic properties than the conventional silicone implant. METHODS: Four miniprostheses (experimental group) and one big silicone implants (control 1 group) were separately implanted beneath the panniculus carnosus muscle of 30 rabbits. After 3 months, measures related to contracture and capsular histology were performed on anesthetized animals. RESULTS: Baker ranking, capsular incision width and capsular thickness of the control groups were evidently higher than that of experimental groups (P < 0.01). Implant compression of the control groups was evidently lower than that of the experimental group. Histology revealed a thinner, more flexed capsule around the miniprostheses as compared with big silicone implants. CONCLUSIONS: The miniprostheses form a looser and thinner capsule than the conventional silicone implant.

Animals↗

[The tangential excision of razor graft for the giant congenital naevi in children].

OBJECTIVE: To study the therapeutical effect of the tangential excision of razor graft for the spacious giant congenital naevi in children. METHODS: The surface layer of the giant naevi, mainly razor graft, was excised tangentially using electrical dermatome. Then the left naevi was removed by dermabrasion. RESULTS: The satisfactory effects were obtained in 9 of 16 cases. The colour of operative area approached the normal skin, occasionally with scattered pigmentation and hypertrophic scars. In 6 cases, the colour appeared lighter than before. One case relapsed after half a year. CONCLUSIONS: The congenital giant naevi in children could be treated using the tangential excision of razor graft. The short-term results might be expected.

Adolescent↗

[A study of incision pattern and pedicles in reduction mammaplasty].

OBJECTIVE: To investigate the efficacy, indication, advantage and disadvantage of methods for reduction mammaplasty. METHODS: Thirty one cases with breast hypertrophy received reduction mammaplasty using various incisions and pedicles. Based on the follow-up of the cases, the appearance, nipple-areola complex sensory function and scars were evaluated. RESULTS: Among the skin incision pattern, inverted T, short-horizontal invert T and vertical incision got excellent breast contour (100%), otherwise, periareolar incision got the less scar (100%). The postoperative result of lateral pedicle, lateral inferior pedicle and mammary gland pedicle were satisfactory in terms of the nipple-areola sensation and the amount of mammary tissue removed. CONCLUSIONS: Breast contour and scar formation postoperative are related to incision design, Invert T incision or short-horizontal invert T incision is a good alternative to moderate and large hypertrophies, Periareolar incision is suitable to small hypertrophies. There is a relationship existing between pedicle location and retention of sensation or amount of mammary tissue removed. The inferior pedicle and lateral-inferior pedicle allows better shaping as well as maintaining good sensation.

Adult↗

[Anatomy study of superior and inferior gluteal artery perforator flap].

OBJECTIVE: To find anatomic basis for clinically modifying technique of harvesting superior and inferior gluteal artery perforator flap, in order to avoid muscle lossing in conventional superior and inferior myocutaneous flaps, keep the advantage such as large rich supplied volume soft tissue. METHODS: 5 cases 10 sides adult cadaver were used to study the numbers, position, Course of superior and inferior gluteal artery perforators. The position of perforators was located by ultrasound Doppler in 6 cases and 12 sides in patient's superior and inferior gluteal area. RESULTS: Superior and inferior gluteal artery originated from internal iliac artery. Several main perforators of large caliber were found in the paraischia and central portions of the gluteal muscle, its number was 10 - 15. The length of the vessels varies from 3 to 8 cm and their diameter from 1 - 1.5 mm. These significant perforators pass through the muscle itself and the fascial portion of the muscle to the overlying skin on the gluteal region. The dorsal branches of nervorum lumbalium perforate the deep fascia just above the iliac crest, lateral to the posterior superior iliac spine. If a nerve branch with a substantial diameter crosses the incision line, the nerve can be harvested within the flap. This nerve can be anastomosed to the anterior ramus of the lateral branch of the 4th intercostals nerve. In adult female, 3 - 5 perforators were located by ultrasound Doppler. They distributed in the triangle area among posterior superior iliac crest, the great trochanter and the coccyx. CONCLUSIONS: The area and diameter of perforators of superior gluteal artery were relatively confirmed. It's possible to harvest the perforator flap without any muscle. It has the advantage of conventional myocutaneous flap with out of its disadvantages. It's easy to detect those perforator by ultrasound Doppler clinically. The nerve can be harvested and anastomosed simultaneously. Because the inferior gluteal area is a weight loading area, we suggested to use superior gluteal artery perforator flap. This flap can be transferred pedicled to treat sacral pressure sores or to be transferred freely for the breast reconstruction.

Adult↗

[The experimental researches on the use of triamcinolone acetonide for the prevention of implant capsular contracture].

OBJECTIVE: To explore the use of triamcinolone acetonide for the prevention of implant capsular contracture. METHODS: 20 rabbits were randomly undivided into 2 groups of 10 animals each. Every 10 ml silicone implant was implanted beneath the panniculus carnosus muscle of one rabbit. At the same time, a modified expander catheter was mounted on the implant. This catheter has many lateral holes and the end was blind. Triamcinolone acetonide (10 mg/3 ml) was infused through the expander pot and catheter as the experimental groups. On the other hand, 3 ml saline was used as the control group at 1, 2, and 3 months. At 6 months, measures related to contracture and capsular histology examinations were performed on anesthetized animals. RESULTS: Baker scores, capsular incision width and capsular thickness of the saline groups were evidently higher than that of triamcinolone acetonide groups (P < 0.01). Implant compression of the saline groups was evidently lower than that of triamcinolone acetonide group. Histology revealed a thinner capsules and less fibrous tissue deposition around the triamcinolone acetonide group, as compared with saline group. CONCLUSIONS: It is effective to deliver triamcinolone acetonide to reduction of capsular contracture through the catheter and its pot.

Animals↗

Silencing of SARS-CoV spike gene by small interfering RNA in HEK 293T cells.

Two candidate small interfering RNAs (siRNAs) corresponding to severe acute respiratory syndrome-associated coronavirus (SARS-CoV) spike gene were designed and in vitro transcribed to explore the possibility of silencing SARS-CoV S gene. The plasmid pEGFP-optS, which contains the codon-optimized SARS-CoV S gene and expresses spike-EGFP fusion protein (S-EGFP) as silencing target and expressing reporter, was transfected with siRNAs into HEK 293T cells. At various time points of posttransfection, the levels of S-EGFP expression and amounts of spike mRNA transcript were detected by fluorescence microscopy, flow cytometry, Western blot, and real-time quantitative PCR, respectively. The results showed that the cells transfected with pEGFP-optS expressed S-EGFP fusion protein at a higher level compared with those transfected with pEGFP-S, which contains wildtype SARS-CoV spike gene sequence. The green fluorescence, mean fluorescence intensity, and SARS-CoV S RNA transcripts were found significantly reduced, and the expression of SARS-CoV S glycoprotein was strongly inhibited in those cells co-transfected with either EGFP- or S-specific siRNAs. Our findings demonstrated that the S-specific siRNAs used in this study were able to specifically and effectively inhibit SARS-CoV S glycoprotein expression in cultured cells through blocking the accumulation of S mRNA, which may provide an approach for studies on the functions of SARS-CoV S gene and development of novel prophylactic or therapeutic agents for SARS-CoV.

Cell Line↗

Accelerating tissue expansion by application of topical papaverine cream.

The rate of tissue expansion can be accelerated by papaverine through a special delivery system, according to an early report. Because the delivery system was complex and inconvenient, another means of administrating papaverine was tested to observe the rate of tissue expansion. In this study, 24 miniature pigs were divided equally into three groups. Four 150-ml silicone expanders were implanted into each pig in groups A and C. Four modified rectangular silicone expanders were also implanted into each animal in group B. During the expansion process, 1 g of 2% hydrochloride papaverine cream was applied topically onto the surface of each pig's expanding skin in group A two times daily, and hydrochloride papaverine solution was injected into the outer shell of each modified expander in group B weekly. Group C acted as the control group. The mean sum of the first four times of saline water volume that was injected into the expanders was 142.42 +/- 5.6 ml in group A, 128.72+/-4.8 ml in group B, and 106.38+/-3.28 in group C. There were statistical differences among the three groups. The mean sum of volume saline water that was injected into hind expanders was 137.51 +/- 5.1 ml in group A, 120.35 +/- 3.6 ml in group B, and 102.63 +/- 4.76 ml in group C, and there was a statistical difference among the three groups as well. There was no statistically significant difference in the thickness of fibrous capsules among the three groups. This study shows that the rate of tissue expansion can be accelerated by topical application of papaverine cream, and the rate is better than the rate of tissue expansion induced by the special drug delivery system.

Administration, Topical↗

[Computer-assisted simulation and clinical application of custom-produced nose implant].

OBJECTIVE: Surgery to reshape the nose with an implant has been a regular procedure for enhancing a patient's appearance and self-confidence. The purpose of this study was to establish a three-dimensional surgical simulation system. which can provide the patients with realistic prediction of their own postoperative appearance in computer and specifically produce a nose implant for an individual patient. METHODS: Preoperative CT data and image-processing techniques were employed to generate a three dimensional template of the papatient' s face. Three-dimensional morphing processes were then applied to predict and compare the outcome of plastic procedures on a patient-specific nose surface, according to the patient' s expectation. By comparing and revising the changes before and after simulation, digital prototypes of the nose implants could be generated. The templates to create custom-made silicone implants were than produced by a computer controlled device. RESULTS: Accurately regeneration of 3-D images and realistic operative simulations could be achieved with this system. The implants produced exactly conformed to the results of simulation. No curving and reshaping were needed during operating. The clinical results extremely matched with the simulations. CONCLUSION: The system enhances surgeon-patient communication and facilitates preoperative planning. It is especially desirable for implant surgery with less guesswork of size, contour, and orientation of the implant. The best chance of optimal results could be achieved.

Adolescent↗

[Breast reconstruction with multiple miniprostheses].

OBJECTIVE: To evaluate a new technique for breast reconstruction with multiple miniprostheses, METHODS: Each silicone miniprosthesis(10 approximately 15 ml) was implanted one by one beneath the prepared muscle pocket until the desired volume was achieved. RESULTS: Seven patients were treated with the above mentioned technique, including five after gland fibroadenoma excision, one hemangioma excision and one reduction mammoplasty. The results were satisfactory. CONCLUSION: The above mentioned technique with the miniprostheses may be another good way for breast augmentation and breast reconstruction with many advantages such as smaller incision, easy intraoperative assessment of the ideal size of the breast, easy handling the technique, perhaps low incidence of capsular contracture.

Adult↗

[Minimizing the antigenicity of tracheal allografts by using a technique of cryopreservation].

OBJECTIVE: To investigate the proper time of cryo-preserving tracheal allograft so as to minimize its antigenicity. METHODS: On a dog model, this study was carried out by allografting a tracheal into a muscular flap formed with sternocephalic muscle and sternohyoid--sternothyroid muscle. The tracheal was treated with cryopreservation in defferent intervals. The viability of the graft was evaluated by the examination of fiberoptic bronchoscopy, histopathology and microangiography. The blood flow of the tracheal mucous was measured with a blood flowmeter and the survival area was decided in the calculation of the percentage. RESULTS: There are no significant differences in the mucous membrane appearance and the mucosal blood flow one week after the surgery among the non-cryopreservation group and the groups treated with cryopreservation in 1 day, 2 weeks, 4 weeks, 6 weeks and 8 weeks. The graft was found to start necrosis 2 weeks after the transplantation with the infiltration of mononuclear cells examined under light microscope in almost all of the groups, especially in the non-cryopreservation group and the groups treated with cryopreservation in 1 day, 2 weeks. However, there was no significant difference among the autograft group and the allograft groups cryopreservated in 6 weeks and 8 weeks, and the infiltration of the mononuclear cells was not found in these groups either. CONCLUSION: The antigenicity of the tracheal allografts could be significantly decreased by the treatment of cryopreservation over 6 weeks.

Animals↗

[Experimental study on inhibition of skin graft contraction with papaverine cream].

OBJECTIVE: To evaluate the effect of exteral use of papaverine cream on enhancement of skin graft. METHODS: Five mini-pigs were used for the study. 20 full-thickness wounds were created on the back of each animal. Along the midline of the abdomen, a 40 cm x 2 cm full-thickness skin graft was harvested and the wound sutured.The full-thickness graft was prepared and trimmed to 2 cm x 2 cm of 0.6 mm thickness split-skin grafts, which were transplanted to each wound with tie-over bolsters. The sutures were removed 2 weeks after the operation. Twelve pairs of 100%-survived skin grafts were selected and divided into group A (the left side) and group B (the right side), with each pair locating on the same and opposite position. From the day of suture removal, 2% papaverine cream was used to group A while plain cream was used to group B. The grafts were measured and observed for healing condition at the time of suture removal and the first, second, third, fourth, fifth, and sixth month. The surface area of the graft was measured and the shrinking ratio was calculated. After the animals were killed at the sixth month, samples of the skin grafts were taken and processed with 10% formalin, routine paraffin wax and Hematoxylin-eosin staining. The tissue structure was observed and the results were analyzed statistically. RESULTS: The grafts in two groups did not have significant differences at the time of suture removal. Observations from the first to the sixth month showed that the two groups had significant differences in skin graft contracture and histological changes. HE stains demonstrated that the skin grafts in group A had less fibroblasts, more microvessels and orderly-arranged collagenous fibers. CONCLUSIONS: External use of papaverine cream can inhabit the contraction of skin grafts. It is a safe, effective, simple, and reliable method,which has the advantages of easy delivery,cheapness, less injury and infection,and no side effects.

Animals↗

Antigen-expressed recombinant Salmonella typhimurium driven by an in vivo-activated promoter is capable of inducing cellular immune response in transgenic mice.

To explore the approaches and mechanisms for reversing the immune tolerance in transgenic mouse, and the pathogenicity of hepatitis G virus (HGV), the promoter of phoP-activated gene (P(pagC)) of Salmonella typhimurium was used as a transcriptionally regulating element to construct an attenuated S. typhimurium expressing HGV NS3. The recombinant S. typhimurium was orally administered to HGV transgenic mice. As the results, HGV antigen in serum and liver as well as HGV mRNA in liver were decreased significantly, although the serum anti-HGV NS3 remained undetectable as the control transgenic mice. The spleen cell proliferation, in vitro HGV NS3 specific CTL, and IFN-gamma assays with the primed cultured splenocytes indicated the induction of Th1 immune responses in those administered transgenic mice. Adoptive transfer of fractionated primed spleen cells to the transgenic mice showed that T lymphocytes were responsible for, maybe through IFN-gamma, the down-regulation of HGV mRNA transcription. Histological examination found no significant inflammatory changes in liver of the transgenic mice. These findings suggested that the oral inoculation of the HGV NS3-expressed attenuated S. typhimurium driven by an in vivo-activated promoter should be a simple and effective approach for potential treatment of chronic viral infection.

Adoptive Transfer↗

[The investigation of tracheas transplantation by wrapping in a muscle flap].

OBJECTIVE: To investigate the possibility of tracheas transplantation by wrapping it in a muscle flap. METHODS: With a dog model, a number of tracheas were separately wrapped in the unilateral sternocephalic muscle flap and the bilateral sternohyoid-sternothyroid muscle flap, and placed in the original site. The tracheas autografting was used as a control. The viability was evaluated by the examination of fiberoptic bronchoscopy, histopathology and microangiography, the measurement of tracheal mucosal blood flow and the calculation of survival rate and percentage of patency. RESULTS: The submucosal blood flow of the transplanted tracheas was detected in the unilateral sternocephalic muscle flap group and the bilateral sternohyoid-sternothyroid muscle flap group 1 week after the surgery and gradually reached the level close to the normal in 4 weeks, while the vascular ingrowth was also shown from the wrapped muscle flap into the transplanted tracheas by using a microangiography technique. The histopathological examination demonstrated that the structure of the transplanted tracheas was quite same as the original one and its inner surface was also covered with pseudostratified columnar ciliary epithelia. However, in the control group, the mucous membranes turned black one week after the transplantation and all dogs died from the graft necrosis. CONCLUSION: The tracheas wrapped in a muscular flap could survive well for a long time.

Animals↗

[Study on single nucleotide polymorphism of TIGR gene in primary open-angle glaucoma patients].

OBJECTIVE: To detect the single nucleotide polymorphism (SNP) of trabecular meshwork-induced glucocorticoid response protein (TIGR) gene and to investigate the association between SNP and primary open-angle glaucoma (POAG) in Chinese Han population. METHODS: High throughput conformation sensitive gel electrophoresis (HTCSGE) and fluorescent labeling automated sequencing method were used to screen and identify the SNPs of TIGR in 82 unrelated patients with POAG. Restriction endonuclease analysis was used to detect the SNPs of TIGR in 150 sex-and age-matched unrelated control subjects without POAG. The frequencies of genotypes and alleles of each SNP between the two groups were compared by chi-square test. RESULTS: A total of six SNPs were identified in TIGR gene with a length of 2172 bp, including all coding regions and part of promoter, introns and 3'UTR region: 1-83(G --> A), G12R, R46X, R76K, IVS2 + 35(A --> G), and T353I. Significant differences were found in frequencies of the genotype TC and the allele T of T353I between POAG patients and control subjects. The frequency of heterozygote TC was 12.20% in POAG patients, significantly higher than that in control subjects (3.33%, kappa(2) = 6.885, P = 0.009, OR = 4.028, OR95% CI = 1.327 approximately 12.223). The frequency of the allele T was 6.10% in POAG patients, significantly higher than that in control subjects (1.67%, kappa(2) = 6.655, P = 0.010). No significant difference was found between the two groups in frequencies of genotypes and alleles of the other SNPs. CONCLUSION: The T353I polymorphism of TIGR gene is associated with POAG in Chinese Han population.

Aged↗

[The autotransplanted tracheas wrapped in united muscle flap of the neck: an experiment].

OBJECTIVE: To investigate the way of revascularization of donator's trachea wrapped in united muscle flap. METHODS: Using fiberoptic bronchoscopy, histopathology and microangiography, we evaluated the tracheal mucosal blood flow, the survival rate, the percentage of patency, and the graft viability of autograft tracheas with varying lengths wrapped in one-sided sternocephalic muscle flap and two-sided sternohyoid-sternothyroid muscle flap and autograft tracheas with the length of 5 rings without wrapped in muscle flap in 32 dogs. RESULTS: In the tracheal autograft wrapped in the united muscle flap group with a length less than 4 centimeters, the submucosal blood flow of graft could be detected by laser blood flowmetry one week after transplantation, and it reached 60% of the normal, which had no significant difference between the place near the site of anastomosis and the middle part of the graft. Dense vessels could be found to grow from the wrapped muscles into the autografted trachea by microangiography. Histopathological examination demonstrated that the structure of the autograft was the same as what it originally was. the inner surface of the autograft was covered with pseudostratified columnar ciliary epithelia, and no necrotic tracheal cartilages were found. Every autograft could survive over long time. However, at 1 week, most mucous membrane in the middle part of the graft with length over 4 cm was in gray or in pale; hyperemia, edema, and haemorrhage were found near the site of anastomosis. Mucosal blood flow measured by laser blood flowmetry in the middle part of the graft was significantly less than that near the site of anastomosis. Malacia, dissolution or granulation hyperplasia occurred in midportion of the major grafts shortly after transplanatation. As for those autografted trachea without wrapping in muscles flap, mucous membranes turned black one week after the transplantation and all dogs died of graft necrosis later. CONCLUSION: One-sided sternocephalic muscle flap and two-sided sternohyoid-sternothyroid muscle flap can provide blood for the graft and the grafted trachea can survive for a long time.

Animals↗