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Biomedical subjects

Xue Zhou

Publications and source records attributed to Xue Zhou.

17 recordsLinked to original sources

Efficient targeted anticoagulant with active RGD motif.

Three anticoagulants combining large peptide recombinant hirudin variants (rHV2-K47) and Arg-Gly-Asp (RGD) motif related to platelet aggregation were generated, i.e. sequences CRFPRGDADPYCE and CNPRGDFRCI were added to the C-terminus of hirudin to obtain RGD-hirudin 1 and 2, respectively, and the sequence RGDSE was inserted between residues 53-54 of hirudin to obtain RGD-hirudin 3. All products exhibited antithrombin and antiplatelet activities, especially IC50 of RGD-hirudin 1 and 2 were much lower with respect to previously reported similar peptides. Our data suggested that RGD-hirudin 1 and 2 would be promising anticoagulants in clinic. Moreover, the triangular structure of active RGD was shown by computer simulation, which might contribute to our understanding on integrin-related peptides and proteins.

Amino Acid Sequence↗

Modification of the Tet-On regulatory system prevents the conditional-live HIV-1 variant from losing doxycycline-control.

BACKGROUND: We have previously constructed a doxycycline (dox)-dependent HIV-1 variant by incorporating the Tet-On gene regulatory system into the viral genome. Replication of this HIV-rtTA virus is driven by the dox-inducible transactivator protein rtTA, and can be switched on and off at will. We proposed this conditional-live virus as a novel vaccine approach against HIV-1. Upon vaccination, replication of HIV-rtTA can be temporarily activated by transient dox administration and controlled to the extent needed for optimal induction of the immune system. However, subsequent dox-withdrawal may impose a selection for virus variants with reduced dox-dependence. RESULTS: We simulated this on/off switching of virus replication in multiple, independent cultures and could indeed select for HIV-rtTA variants that replicated without dox. Nearly all evolved variants had acquired a typical amino acid substitution at position 56 in the rtTA protein. We developed a novel rtTA variant that blocks this undesired evolutionary route and thus prevents HIV-rtTA from losing dox-control. CONCLUSION: The loss of dox-control observed upon evolution of the dox-dependent HIV-1 variant was effectively blocked by modification of the Tet-On regulatory system.

Binding Sites↗

The genetic stability of a conditional live HIV-1 variant can be improved by mutations in the Tet-On regulatory system that restrain evolution.

Live attenuated human immunodeficiency virus type 1 (HIV-1) vaccines are considered unsafe because more quickly replicating pathogenic virus variants may evolve after vaccination. As an alternative vaccine approach, we have previously presented a doxycycline (dox)-dependent HIV-1 variant that was constructed by incorporating the tetracycline-inducible gene expression system (Tet-On system) into the viral genome. Replication of this HIV-rtTA variant is driven by the dox-inducible transcriptional activator rtTA and can be switched on and off at will. A large scale evolution study was performed to test the genetic stability of this conditional live vaccine candidate. In several long term cultures, we selected for HIV-rtTA variants that no longer required dox for replication. These evolved variants acquired a typical amino acid substitution either at position 19 or 37 in the rtTA protein. Both mutations caused rtTA activity and viral replication in the absence of dox. We designed a novel rtTA variant with a higher genetic barrier toward these undesired evolutionary routes. The corresponding HIV-rtTA variant did not lose dox control in long term cultures, demonstrating its improved genetic stability.

AIDS Vaccines↗

Principal axis-based correspondence between multiple cameras for people tracking.

Visual surveillance using multiple cameras has attracted increasing interest in recent years. Correspondence between multiple cameras is one of the most important and basic problems which visual surveillance using multiple cameras brings. In this paper, we propose a simple and robust method, based on principal axes of people, to match people across multiple cameras. The correspondence likelihood reflecting the similarity of pairs of principal axes of people is constructed according to the relationship between "ground-points" of people detected in each camera view and the intersections of principal axes detected in different camera views and transformed to the same view. Our method has the following desirable properties: 1) Camera calibration is not needed. 2) Accurate motion detection and segmentation are less critical due to the robustness of the principal axis-based feature to noise. 3) Based on the fused data derived from correspondence results, positions of people in each camera view can be accurately located even when the people are partially occluded in all views. The experimental results on several real video sequences from outdoor environments have demonstrated the effectiveness, efficiency, and robustness of our method.

Algorithms↗

[Impact of acupuncture on NP-1 expression in spared dorsal root ganglion].

OBJECTIVE: To investigate the impact of acupuncture on NP-1 expression in spared dorsal root ganglion (DRG). METHODS: Twenty-five male adult cats were divided into five groups. The five cats in the pseudo-operation group were kept intact. Ten cats were subjected to unilateral root rhizotomy(unilateral L1-L5 DRGs were transected, but L6 DRG was spared), with five sacrificed seven days later and another five sacrificed 14 days later. Ten cats with unilateral root rhizotomy received acupuncture (electron-needle stimulation in Zusanli, Xuanzhong, Futu and Sanyinjiao which are located in the distribution area of L6 spinal nerve) for 30 minutes per day. Five cats in the acupuncture group were sacrificed seven days and 14 days later respectively. The L6 DRG from the experimental side of each animal was taken and made into frozen sections 20 microm in thickness. The sections were stained under the same conditions using NP-1 specific antibody (1:600, Santa) by immunohistochemistry ABC method. The amounts of NP-1 immunoreaction neurons of DRG were measured. RESULTS: The NP-1 immunoreactants were mainly located in the small and medium-sized neurons and a few large sized neurons in the cats of the pseudo-operation group. Less NP-1 positive medium-small sized neurons were found seven days after partial dorsal root rhizotomy (P<0.05). But they returned back to the original level 14 days after operation. The NP-1 positive large neurons had no significant changes after operations. With seven days of acupuncture, the amounts of NP-1 positive small and medium-sized neurons increased significantly (P<0.05) but were still less than the original level (P<0.05). There were no differences in neurons 14 days after operations between the acupuncture and non-acupuncture group (P>0.05) and between the groups with seven days acupuncture and 14 days of acupuncture (P>0.05). CONCLUSION: Acupuncture increases the expression of NP-1 in small and medium-sized neurons of spared DRG.

Acupuncture↗

[Expression of CNTF and PDGF in spared dorsal root ganglion after partial dorsal root rhizotomy].

OBJECTIVE: To investigate the expression of ciliary neurotrophic factor (CNTF) and platelet derived growth factor (PDGF) in spared dorsal root ganglion (DRG) after partial dorsal root rhizotomy. METHODS: Twenty adult cats were divided into four groups. Five cats were kept intact in the control group. Fifteen cats were subjected to bilateral root rhizotomy, and on the 3rd day, 7th day and 14th day after operation, they were sacrificed as subjects in the three experiment groups respectively (n=5 per group) and their DRGs (L6) were taken. Immunochemical ABC method was used to detect the distribution of CNTF and PDGF-immunoreactivity neurons in those DRGs. The quantitative analysis was conducted to get the numbers of CNTF, PDGF-positive total, large, and medium-small sized neurons in each group. RESULTS: CNTF, PDGF-immunoreactants were distributed in large and medium-small sized neurons. The numbers of total and medium-small sized CNTF positive neurons were noted to be decreased on the 3rd day after operation (P<0.05), and no difference 7th and 14th days was seen when compared with control (P>0.05), but the large positive neurons showed no difference. The total and medium-small sized PDGF positive neurons were found decreased apparently on the 3rd day and 7th day, but there was no difference on the 14th day as compared with normal level. Large positive neurons displayed no change at every time. CONCLUSION: Partial dorsal root rhizotomy exerts different influence on the expression of CNTF and PDGF for different neurons in spared DRG.

Animals↗

[Expression of Bax, Bcl-2 and caspase-3 in spared dorsal root ganglion after partial dorsal root rhizotomy].

OBJECTIVE: Investigating the expression of Bax, Bcl-2 and Caspase-3 in dorsal root ganglion after partial dorsal root rhizotomy in spared root. METHODS: Adult cats were used. The study comprised a normal control group (n=5) and three expriment groups (n=5 per group). The unilateral root rhizotomy was performed on the 15 cats of the 3 experiment groups. On the 3rd, 7th and 14th days after partial dorsal rhizotomy, the DRG (L6) of 5 cats per batch were taken and made into frozen sections and were stained under the same condition using specific Bax, Bcl-2 and Caspase-3 antibody by ABC method. The Bax, Bcl-2 and Caspase-3 positive large-, medium- and small-sized neurons in DRG were counted in the 3 experiment groups and in the normal control group. RESULTS: In the intact animals of the normal control group, the Bax, Bcl-2 and Caspase-3 immunoreactants were mainly distributed in medium, small neurons and a few large neurons. The number of Bax positive neurons in spared DRG of the 3rd day group increased apparently as compared with that of the normal control group but decreased apparently in the 14th day group and showed no significant difference when compared versus control (P>0.05). There was no significant difference between the 14th day group and normal control group. Bcl-2 expression in spared DRG displayed no difference among the four groups. The number of Caspase-3 positive neurons in spared DRG of the 3rd day group increased apparently when compared against control (P<0.05), but it was not significantly different from that of the 7th day and 14th day groups (P>0.05). CONCLUSION: Partial dorsal root rhizotomy mainly has an effect on the expressions of Bax, Bcl-2 and Caspase-3 in medium- and small-sized neurons of spared DRG, the peaks of the expressions may appear 3-7 d after the operation.

Animals↗

[Changes in expression of SP in spinal lamina II following hemisected spinal cord injury in rats].

OBJECTIVE: To explore the changes in expression of Substance P (SP) in spinal lamina II at different time following hemisected spinal cord injury (hSCI). METHODS: Twenty Sprague-Dawley rats were randomly divided into intact group and hSCI group at days 3, 7 and 21. The spinal cords were hemisected between T13 and L1. The animals were sacrificed 3, 7, 21 days after operation. The L3 segments were taken out and sectioned continuously into sections (20 microm). The expression of SP was measured by immunohistochemical ABC method, and the number of SP positive varicosities in lamina II was counted. RESULTS: SP positive varicosities were observed in spinal lamina II. Compared with control group, the number of SP positive varicosities on the injured side apparently increased 3 days after hSCI, but decreased 7 days after operation, then recovered to normal by 21 days after operation. Noteworthily, the number of SP positive varicosities on the intact side was higher than that on the injured side. CONCLUSION: SP, as a neurotransmitter of nociceptive information, may be related to the process of hSCI.

Animals↗

[Distribution of epidermal growth factor in central nervous system of adult rhesus monkey].

OBJECTIVE: To investigate the distribution of epidermal growth factor (EGF) in the CNS of adult rhesus monkey. METHODS: Frozen sections were incubated in specific polyclonal anti-EGF antibody by the immunohistochemical SP method. RESULTS: The EGF immunopositive reaction was observed in the plasma of neurons. The neurons with strong positive immuno-reaction signals were detected in cerebral cortex, cerebellar Purkinje cells, cerebellar nuclei, pyramidal neurons of hippocampus, caudate nucleus, lentiform nucleus, claustrum, nuclei in diencephalons, substantia nigra, cranial nerve nuclei, reticular formation in brain stem, pontine nuclei, red nucleus, superior and inferior olivary nucleus, gracile nucleus, cuneate nucleus, also the ventral horn, lateral horn, dorsal horn and the central gray matter in spinal cord. Furthermore, a few EGF immunopositive glias and fibres were observed in some white matter of central nervous system. CONCLUSION: EGF immunoreactive material was extensively distributed in the CNS of the adult rhesus monkey. The results suggest that EGF may be concerned with various types of neurons and other cells.

Animals↗

[Effect of partial ganglionectomy and acupuncture on culturing spared DRG in vitro].

OBJECTIVE: To explore the effect of partial dorsal root rhizotomy and Acup on culturing dorsal root ganglion(DRG) in vitro. METHODS: Ten adult cats were divided into 2 groups: normal control group; Acup spared DRG 7 d group, in which bilateral L1-L5, L7-S2 DRG were removed; and L6DRG were spared; then unilaterally two sets of acupoints [Zusanlily (St. 36) and Xuanzhong (G. B. 39): Futu (St. 32) and Sanyinjiao (Sp. 6) located in the distribution area of spinal nerve L6] were electro-stimulated alternatively 30 min everyday by electro-needling. Five cats were used in every group. Bilateral L6 DRGs of every group were taken out on the condition of asepsis and were cultured respectively in vitro. Cultures were terminated after day 7. Then the cultured cells were stained under the same condition using specific NSE (1 : 200) antibody, a neuron-specific marker, by the immunohistochemistry ABC method. The neurite length was measured by micro-measured ruler in upside-down light microscope on the 1st, 3rd, 5th, 7th day. RESULTS: Immunocytochemical staining revealed that over 95% cells were NSE positive cells which were the typical neuron of DRG in vitro; on the 1st, 3rd, 5th, 7th day, the average neurite length of the normal group was shorter than that of the spared DRG group(P < 0. 05), and the spared DRG group's was shorter than the Acup group's at each time stage (P < 0.05). CONCLUSION: These results indicated that DRG had plasticity and acupuncture probably promoted the plasticity, which were probably in close relation with the spinal plasticity.

Animals↗

Viral evolution as a tool to improve the tetracycline-regulated gene expression system.

We present viral evolution as a novel and powerful method to optimize non-viral proteins. We used this approach to optimize the tetracycline (Tc)-regulated gene expression system (Tet system) for its function in mammalian cells. The components of the Tet system were incorporated in the human immunodeficiency virus (HIV)-1 virus such that viral replication is controlled by this regulatory system. Upon long term replication of this HIV-rtTA virus in human T cells, we obtained a virus variant with an enhanced replication potential resulting from an improved rtTA component of the introduced Tet system. We identified a single amino acid exchange, F86Y, which enhances the transcriptional activity and doxycycline (dox) sensitivity of rtTA. We generated a new rtTA variant that is 5-fold more active at high dox levels than the initial rtTA, and 25-fold more sensitive to dox, whereas the background activity in the absence of dox is not increased. This new rtTA variant will be very useful in biological applications that require a more sensitive or active Tet system. Our results demonstrate that the viral evolution strategy can be used to improve the activity of genes by making them an integral and essential part of the virus.

Biotechnology↗

[The neurotrophic effect of endogenous NT-3 from adult cat spared dorsal root ganglion on ganglionic neurons].

OBJECTIVE: To investigate the neurotrophic effect of endogenous NT-3 from adult cat dorsal root ganglion (DRG) on ganglionic neurons. METHODS: Rhizotomy of bilateral L1, L3, L5 and L7 dorsal roots of cats was performed, leaving L2, L4 and L6 DRG as spared DRGs. The separate neurons of normal (control) DRG, spared DRG and anti-NT-3 antibody blocking DRG were cultured in vitro respectively. The number of survival neurons and the length of neurites were measured and used for comparison in the control, spared DRG, and block groups. RESULTS: There were survival neurons and cell clusters in every group. The number of survival neurons and cell clusters of spared DRG group were much larger than those of the control and block groups. The neurite length of neurons, the neurite number and the length of cell clusters of spared DRG group were much greater than those of control and block groups. CONCLUSION: Endogenous NT-3 from spared DRG may act on ganglionic neurons to maintain survival of neuron and stimulate growth of neurite.

Animals↗

[Expressional change of nitric oxide synthases in dorsal root ganglia of cats after selective dorsal rhizotomy].

OBJECTIVE: To examine the expressional change of nitric oxide synthase (NOS) in the injured dorsal root ganglia (DRG) and the ipsilateral adjacent uninjured DRG after selective dorsal rhizotomy. METHODS: Immunochemical ABC method was used to detect the distribution of immunoreaction complex of NOS isoforms--nNOS and eNOS, and quantitative analysis was conducted to get the number of nNOS-immunoreactivity (nNOS-IR) neurons in normal DRG, dorsal rhizotomized DRG and spared DRG from adult cats on the 6th day after operation. This operating model was made by rhizotomizing unilateral L1-L5 dorsal roots and leaving L6 as a spared root. RESULTS: nNOS-immunoreactants were mainly distributed in the small-sized neurons in the DRG of cat. The percentage of nNOS-expressing small-sized neurons increased in the deafferentated L5 DRG (29.74%) when compared with the contralateral DRG (19.35%), and it also increased in the spared DRG (24.22%), compared with the contralateral DRG (18.61%). eNOS-IR was not observed in the DRG of adult cats. CONCLUSION: nNOS/NO up-regulated in DRG neurons is involved in a wide variety of biological functions under physiological and lesion-induced pathophysiological conditions in nerve system.

Animals↗

Conditional virus replication as an approach to a safe live attenuated human immunodeficiency virus vaccine.

Despite intensive efforts, no safe and effective vaccine has been developed for the prophylaxis of human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS). Studies with the simian immunodeficiency virus (SIV)/macaque model demonstrated that live attenuated viruses are the most effective vaccines tested thus far. However, due to ongoing low-level replication of the attenuated virus and the error-prone replication machinery, the attenuated virus may regain replication capacity and become pathogenic. We therefore designed a novel vaccine strategy with an HIV-1 virus that replicates exclusively in the presence of the nontoxic effector doxycycline (dox). This was achieved by replacement of the viral TAR-Tat system for transcriptional activation by the Escherichia coli-derived Tet system for inducible gene expression. This designer HIV-rtTA virus replicates in a strictly dox-dependent manner and may represent an improved vaccine strain because its replication can be turned on and off at will. Spontaneous virus evolution resulted in optimization of the components of the Tet system for their new function to support virus replication in human cells. The optimised Tet system may be of particular use in other applications such as inducible expression of gene therapy vectors in the brain.

AIDS Vaccines↗

Conditional live virus as a novel approach towards a safe live attenuated HIV vaccine.

To control the worldwide spread of HIV, a safe and effective prophylactic vaccine is urgently needed. Studies with the simian immunodeficiency virus demonstrated that a live attenuated virus can be effective as a vaccine, but serious concerns about the safety of such a vaccine virus have arisen. We propose a conditional live virus, of which the replication can be switched on and off at will, as a novel approach for an HIV vaccine.

AIDS Vaccines↗

[The spatiotemporal change of neurotrophin family and their mRNA expression in spinal cord of cats after partial rhizotomy].

OBJECTIVE: To investigate the spatiotemporal change rule of NGF, BDNF, NT-3 and their mRNA expression in spinal cord of cats after partial dorsal rhizotomy. METHODS: Rhizotomy of unilateral L1-L5, L7-S2 dorsal roots of cats was performed, leaving L6 as a spared dorsal root. By using ABC immunohistochemistry and in situ hybridization techniques, the dynamic changes of the above three factors and their mRNA in spinal lamina II of different segments were analysed. RESULTS: 1. In normal cat, NGF and it's mRNA were detected in part of neurons; BDNF, in nerve fiber terminals, varicosities and neurons; NT-3, in part of neurons, neuroglias and few nerve fiber terminals and varicosities. The mRNAs of the later two were negative. 2. The population of NGF and NGGmRNA positive neurons, NT-3 positive neurons and neuroglias increased significantly 3d-5d after rhizotomy. However, the quantity and density of positive varicosity of BDNF decreased. At 10-11d, the population of NGF and NGF mRNA positive neurons was still on the high level as that at 3-5d, and that of NT-3 began to decrease; the quantity of BDNF recovered to normal except for L, segment, but the density of positive varicosity of BDNF did not yet. The mRNAs of BDNF and NT-3 were still negative. 3. The change of each factor varied with the segments. The highest level time of NGF was earlier in L5, L6 than in L3; the recovery of the quantity of BDNF was the latest in L7; the change of NT-3 positive neuroglia was the same at each segment, but the number of NT-3 positive neuron in L5, L7 returned to normal at 10-11d, and that of L3 did not. CONCLUSION: The three factors all play roles in spinal plasticity after partial rhizotomy, but they function at different time phase and last different time length.

Animals↗

[Effect of acupuncture on the expression of NT3 in the process of spinal plasticity].

OBJECTIVE: To explore the change in the expression of NT3 in the process of promoting the plasticity of spinal cord by acupuncture. METHODS: Five adult cats were subjected to unilateral spared root rhizotomy; their L1-L5, L7-S2 dorsal root ganglia (DRG) were sectioned, but L4 was spared. And two groups of acupoints [Zusani (St.36) and Xuanzhong (G. B.39); Futu (St.32) and Sanyingjiao (Sp.6)] located in hind limb were electro-stimulated for thirty minutes q.d. x 7. At seven days, after acupuncture, the L5 segment of spinal cord and spared dorsal root ganglion (L6) were taken and made into frozen section 20 microns in thickness. Immunohistochemistry (NT3 antibody 1:1500) and in situ hybridization (NT3 cRNA probe 1:100) techniques were used. The numbers of positive neuron for NT3 and it's mRNA in large, medium, small neuron of L6 DRG and the numbers of positive neurons and glia cells for NT3 in lamina II were counted respectively. RESULTS: The numbers of positive large, small neurons for NT3 and its mRNA in DRG and the number of positive neurons and glia cells for NT3 in lamina II on the acupuncture side increased apparently than those on the non-acupuncture side (P < 0.05). However, the positive signal of NT3 mRNA in lamina II was not seen in our study. CONCLUSION: The results indicate that acupuncture promoting the plasticity of spinal cord involves both the increase in expression of NT3 in large and small neurons of spared DRG and the increase in number of NT3 positive neurons and glia cells in spinal lamina II. Moreover, NT3 may play a role in the process of promoting the plasticity of spinal cord by acupuncture.

Animals↗