Analysis of Human Herpes Viruses-Activated Infection Spectra in Patients with Various Immunodeficiencies / 中国实验血液学杂志

OBJECTIVE@#To study the epidemiologic characteristics of human herpes virus (HHV) activated infection in the diseases of blood system and patients received allo-HSCT by statistically analyzing the screening results of 8 human herpes viruses (HHVs) of 4164 patients in Hebei Yanda LU Dao-Pei Hospital from 2012 to 2017.@*METHODS@#PCR was used to screen 8 HHVs.@*RESULTS@#Two thousand and fifty-two patients (49.28%) were HHV-positive among 4164 patients screened. Among these patients screened, the infection spectra of 8 human HHVs in hematological diseases as well as patients received allogeneic hematopoietic stem cell transplantation of totally 2994 patients were summarized as follows: the positive rate of EBV (29.49%) was the highest, that of HCMV (23.15%), HHV-6 was 18.77% and HHV-7 was 17.64%, while the remaining 4 HHVs all≤2.1%. The rate of co-infection of various HHVs was significantly higher than that of single infection of HHV among all these disease groups except familial hemophagocytic lymphohistiocytosis, for which single EBV infection was the most common. The differences of positive rates among these 8 human HHVs in hematological diseases as well as patients received allogeneic hematopoietic stem cell transplantation were statistically significant by Chi-square test of R*C tables (χ=54.99, P＜0.05). For each HHV, the differences of positive rates among the above-mentioned disease groups were also statistically significant except HHV-8 (P＜0.05).@*CONCLUSION@#The patients with various blood diseases have different activated infection spectra of HHVs. EBV, HCMV, HHV-6 and HHV-7 are most common in HHVs infection. Different HHVs infections correlate with different hematologion diseases.

Effect of Aconiti Lateralis Radix Praeparata Polysaccharide on Expressions of MMP-2,MMP-14 in Gastric Cancer Xenografts in Nude Mice / 中国实验方剂学杂志

Objective: To observe the growth inhibition effect of Aconiti Lateralis Radix Praeparata polysaccharide on the gastric cancer xenografts in nude mice and the expressions of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-14 (MMP-14) in tumor tissues. Method: The nude mice xenograft model was established and randomly divided into model group, 5-fluorouracil group (5-FU, 0.01 g · kg-1), high and low-dose Aconiti Lateralis Radix Praeparata polysaccharide groups (0.2, 0.1 g · kg-1). Each group was given drug by gavage for 15 days. The effect of Aconiti Lateralis Radix Praeparata polysaccharide on the weight of gastric cancer in nude mice was observed. Morphological changes of tumor cells were observed under light microscope. The content of transforming growth factor-β1 (TGF-β1) in serum was determined by enzyme-linked immunosorbent assay (ELISA). The protein and mRNA expressions of MMP-2, MMP-14 in tumor tissues were detected by immunohistochemistry, Western blot and Real-time PCR. Result: The tumor inhibition rates of high and low-dose Aconiti Lateralis Radix Praeparata polysaccharides were 52.83%,40.57%, respectively. Compared with model group, the tumor weight of high and low-dose Aconiti Lateralis Radix Praeparata polysaccharide was significantly lower (Pβ1 content in nude mice (PPPPPPPConclusion: Aconiti Lateralis Radix Praeparata polysaccharide can inhibit the growth of gastric cancer xenografts in nude mice and the expression of TGF-β1.The anti-tumor mechanism may be related to the down-regulation of MMP-2, MMP-14 protein and mRNA expressions.

Link between sortase A function and cariogenicity of Streptococcus mutans: a preliminary metabolomics analysis / 华西口腔医学杂志

<p><b>OBJECTIVE</b>This study intends to explore the mechanism underlying the support of sortase A (SrtA) of the cariogenicity of Streptococcus mutans (S. mutans).</p><p><b>METHODS</b>We performed a metabonomics study based on ¹H nuclear magnetic resonance spectroscopy (NMR), in which we compared the extracellular metabolites of wild-type S. mutans UA159 with those of its SrtA-deficient strain. Metabolite differences among strains were identified using a combination of principal component analysis and orthogonality partial least square discriminant analysis.</p><p><b>RESULTS</b>Several differences corresponding mostly to unknown metabolites were identified. Some amino acids such as leucine and valine (δ 0.92×10⁻⁶-1.20×10⁻⁶), lactic acid ( δ1.28×10⁻⁶), oxoglutaric acid (δ 3.00×10⁻⁶), and glycine (δ 3.60×10⁻⁶) differed among strains.</p><p><b>CONCLUSIONS</b>This work establishes the feasibility of using ¹H NMR-based metabonomics to provide leads for research into molecular factors that promote caries. The database of microbial metabolites should be also improved in further studies.</p>

Investigation of awareness of menopausal hormone therapy among medical staff in Zhujiang Hospital of Southern Medical University / 解放军医学杂志

Objective To investigate the awareness and attitude towards menopause and menopausal hormone therapy (MHT) among the medical staff of Zhujiang Hospital of Southern Medical University.Methods A self-designed questionnaire related to menopause and MHT was conducted among 1143 medical staffs in Zhujiang Hospital of Southern Medical University.Results The best-known symptoms,in sequences,were dysphoria and depression (90.6％),sleep disorders (81.5％),hot flashes and night sweating (69.4％),dizziness and palpitation (59.3％),and paresthesia (50.3％).Of 1143 respondents,42.1％(481) knew about MHT,and 62％(709) considered that MHT is necessary for symptomatic menopausal women.Significant differences existed in attitudes towards MHT between different titles and departments (P=0.027,P=0.000).Fifty-seven percent (651) of medical staff expressed concern about the side effects of MHT and had scruples about its use,73.1％(836) believed that MHT can improve menopausal symptoms,while 54.5％(623) believed MHT can prevent and treat osteoporosis.Conclusions The awareness rate on menopause and MHT is relatively low among the medical personnel of Zhujiang Hospital of Southern Medical University.There exist differences in attitudes towards menopause and MHT among different departments,doctors and nurses,and different titles.

Effect of allogeneic adipose derived mesenchymal stem cells on regulatory T cells in peripheral blood of patients with premature ovarian failure / 解放军医学杂志

Objective To investigate the effect of allogeneic adipose derived mesenchymal stem cells (hADSCs) on regulatory T cells (Tregs) in peripheral blood of patients with premature ovarian failure (POF).Methods hADSCs were isolated from adipose tissue using 0.1％ type Ⅰ collagenase digestion.Adipogenic,osteogenic differentiation and surface molecular characterization were also performed.Peripheral blood mononuclear cells (PBMCs) of POF patients were isolated by density gradient centrifugation.PBMCs were co-cultured with 1 × 104,2 × 104 and 1 × 105 hADSCs for 72 hours under the stimulation of phytohemagglutinin (PHA).The proliferation rate of lymphocytes was measured by CCK-8 method,the proportion of CD4+ CD25+ Foxp3+ Treg cells was measured by flow cytometry.Real-time fluorescent quantitative PCR was used to detect the mRNA level of Foxp3+.Results hADSCs were positive for CD90,CD105 and negative for CD34,CD45,and has adipogenic and osteogenic differentiation ability.The results of CCK-8 showed that hADSCs could significantly inhibit the proliferation of lymphocytes compared with that in control group (P＜0.001).Flow cytometry showed that hADSCs could promote the proliferation of CD4+ CD25+ Foxp3+ Treg cells (P＜0.05).The results of qPCR showed that Foxp3+ mRNA expression was obviously up-regulated in all experimental groups compared with that in control group (P＜0.001).Conclusion hADSCs can play an immune regulatory role and promote the proliferation of Tregs in peripheral blood of patients with premature ovarian failure.

Therapeutic effect of lentivirus-mediated miR-21 on chemotherapy-induced premature ovarian failure in rats / 解放军医学杂志

Objective To explore the therapeutic potential of miR-21 in rat model of chemotherapy-induced premature ovarian failure (POF).Methods Lentivirus-mediated miR-21 (LV-miR-21) was constructed successfully in vitro with molecular biology methods.Rats were divided into 4 groups named control group,model group,blank vector group and miR-21 group.Rat models of chem0therapy-induced POF were established in the latter 3 groups by intraperitoneal injection ofcytoxan (CTX).Bilateral ovaries of rats in miR-21 group were injected with LV-miR-21,of rats in blank vector group were injected with lentivirus vector,and rats in model group received no treatment.At 1,15,30,45 and 60 days after the last injection,blood sample was collected,the rats were then sacrificed and the ovaries were removed.The estrous cycle was observed by vaginal smears.The E2 level was detected by chemiluminescent immunoassay,and the follicle stimulating hormone (FSH) level was detected by homologous double antibody radiation immunoassay.Ovary weights were measured,and the follicle count was conducted through observing paraffin section under microscope.The apoptosis of ovarian granulosa cells was analyzed by TUNEL assay.Results During 15-30 days,30-45 days and 45-60 days after the last injection,regular estrous cycle was recovered respectively in 8,5 and 3 rats in miR-21 group.At the 15th,30th,45th and 60th day after the last injection,the E2 level was higher in miR-21 group than in model group and blank vector group,but the FSH level showed the opposite trend (P=0.000).At the 45th and 60th day after the last injection,the follicle numbers at all stages increased markedly in miR-21 group than in model group and blank vector group (P=0.000).At the 30th,45th and 60th day after the last injection,the ovary weights were higher in miR-21 group than in model group and blank vector group.At the 15th,30th,45th and 60th day after the last injection,the apoptosis rate of ovarian granulosa cell were significantly lower in miR-21 group than in model group and blank vector group (P=0.000).Conclusion Up-regulation of miR-21 expression may partly recover the ovarian structure and function damaged by CTX.

Resource investigation of wild Codonopsis pilosula in Tanchang county of Gansu / 中国中药杂志

Tanchang county is the distribution of wild medicinal plant resource-rich region, in order to ascertain Tanchang county Codonopsis pilosula wild resources and reserves of the status quo, according to the fourth national Chinese medicine resources survey technology solutions, using sets of plots and investigating combined route survey method, the county wild C. pilosula var. modesta and C. pilosula resources were investigated by a comprehensive survey designed to reveal the distribution of the county's wildlife resources and herbs C. pilosula reserves. The results showed that in Tanchang county seven ecological zones 53 plots, wild C. pilosula distributed in there were 6 ecological zones 11 plots, accounting for 85.71% of the survey area, wild C. pilosula var. modesta was found only in an ecoregions plots, overlapping with C. pilosula region, accounting for 14.29% of the survey area. C. pilosula herbs reserves were calculated as about 461.85 t, economic capacity of 254.02 t, annual amount of acceptance 25.40 t. C. pilosula var. modesta herbs reserves were calculated as 67.75 t, economic capacity of 36.16 t, acceptance annual amount 3.62 t. The total ash C. pilosula was 3.25%, alcohol-soluble extract was 63.86%, while the C. pilosula var. modesta total ash was 3.69%, alcohol-soluble extract was 68.32%. C. pilosula is suitable for broad range, but wild resource is scarce, C. pilosula var. modesta is suitable for relatively narrow scope, and wild resource is scarce, it is recommended to strengthen the protection of wild resources and the rational development and utilization.

Biological features and ultrastructure of human umbilical cord mesenchymal stem cells / 中国医学科学院学报

<p><b>OBJECTIVE</b>To isolate and culture human umbilical cord mesenchymal stem cells (MSCs) and explore their biological features and ultrastructure.</p><p><b>METHODS</b>After isolating MSCs from the human umbilical cord, the proliferation, cycle, and apoptosis were observed. The cell ultrastructure was observed under transmission electron microscope. The cytokines including vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), and insulin-like growth factor-1 (IGF-1) were detected using enzyme-linked immunosorbent assay.</p><p><b>RESULTS</b>Human umbilical cord MSCs had fibroblast-like morphology and increased proliferation capability. Ultrastructural analysis showed that the MSCs had active cellular metabolism and strong migration and differentiation capabilities. Meanwhile, they could secrete anti-apoptotic cytokines such as VEGF, IGF-1, and HGF.</p><p><b>CONCLUSION</b>Human umbilical cord MSCs can secrete many anti-apoptotic cytokine and have good biological features.</p>

Inhibitory effect of short hairpin RNA targeting survivin gene on human ectopic endometrial cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a lentiviral expression vector for short hairpin RNA (shRNA) of human survivin gene, and assess its gene silencing effect in human ectopic endometrial cells.</p><p><b>METHODS</b>Human survivin gene shRNA sequence was designed using a software available on-line. The synthesized shRNA sequence was cloned into the pGCL-GFP vector to construct LV-survivin shRNA, which was confirmed by PCR and DNA sequence analysis. The packaging 293T cells were cotransfected with LV-survivin shRNA, pHelper 1.0 and pHelper 2.0, and the titer of the lentivirus was determined. The recombinant lentivirus was injected into human ectopic endometrial cells and the survivin mRNA expression was examined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) in comparison with that in the non-transfected and blank vector-transfected human ectopic endometrial cells.</p><p><b>RESULTS</b>PCR analysis and DNA sequencing confirmed correct insertion of the shRNA sequence into the lentiviral vector. The titer of virus after packaging was 8x10(8) U/ml. Survivin mRNA expression in human ectopic endometrial cells transfected by LV-survivin shRNA was significantly inhibited compared with those in the non-transfected and empty vector transfected human ectopic endometrial cells (P<0.01), and no significant difference was found between the latter two groups.</p><p><b>CONCLUSION</b>The lentiviral shRNA vector of survivin gene constructed can effectively inhibit the expression of survivin gene in human ectopic endometrial cells in vitro. This vector provides a tool for investigating the role of survivin gene in the occurrence and progression of endometriosis and for searching new therapeutic targets.</p>

Small interference RNA targeting nuclear factor-kappaB inhibits endometriotic angiogenesis in chick embryo chorioallantocic membrane / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of small interference RNA (siRNA) targeting nuclear factor-kappaB (NF-kappaB) on endometriosis.</p><p><b>METHOD</b>The eutopic endometrium of women with endometriosis were transplanted into the nonvascular region of 8-day-old chicken embryo chorioallantocic membrane (CAM), and the effects of NF-kappaB p65 siRNA on the vascularization and endometriotic lesion formation were tested with proper controls.</p><p><b>RESULTS</b>Transplantation of the endometrium onto the CAM resulted in a strong angiogenic response in the chicken tissue. The angiogenesis was significantly reduced and endometriotic lesion formation significantly suppressed with siRNA targeting NF-kappaB in comparison with the control group.</p><p><b>CONCLUSIONS</b>The NF-kappaB pathway is involved in the development of endometriotic lesions in vitro, and NF-kappaB gene silencing reduces endometriotic angiogenesis and promotes cell apoptosis in the endometriotic lesions, suggesting that NF-kappaB might be a good target for endometriosis treatment.</p>

Protective effect of estrogen against chemotherapy-induced ovarian damage in rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the protective effect of estrogen against cyclophosphamide (CTX)-induced ovarian failure in female rats.</p><p><b>METHODS</b>Sixty female Wistar rats (2-3 months old) were randomized into 4 groups to receive treatments with normal saline, CTX, estradiol (E2) or CTX+E2. Serum estradiol and follicle-stimulating hormone (FSH) concentrations were measured regularly in the 4 groups, and the weight of the ovary and uterus, follicle number and mean diameter of the follicles were compared.</p><p><b>RESULTS</b>Compared with CTX+EE2 group, the CTX group showed significantly increased FSH levels and decreased EE2 levels (P<0.05). The weight of the ovary and uterus and follicle number were significantly lower in CTX group than in CTX+EE2 group (P<0.05). No obvious differences in the indexes were found between the control and CTX+E2 groups.</p><p><b>CONCLUSION</b>Estrogen administration provides protection against CTX-induced ovarian damage in rats.</p>

Construction of a lentiviral vector carrying human bcl-2 gene and its expression in human ovarian granulosa cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a lentiviral vector carrying human bcl-2 gene and investigate its expression in human ovarian granulosa cells (GCs).</p><p><b>METHODS</b>Human bcl-2 gene was amplified from the plasmid pCMV-SPORT6 using PCR and subcloned into the lentiviral vector pGC-FU to construct the lentiviral expression vector pGC-FU- bcl-2. The bcl-2 gene insert was confirmed by restriction enzyme digestion and sequencing. The recombinant lentiviruses generated by 293T cells co-transfected with pGC-FU-bcl-2 and the packaging plasmids pHelper1.0 and pHelper2.0. The resulting recombinant lentivirus GC-FU-bcl-2 carrying bcl-2 and EGFP genes were then used to infect human ovarian granulosa cells. EGFP and bcl-2 protein expressions in 293T and human ovarian GCs were detected by fluorescent microscope and Western blotting.</p><p><b>RESULTS</b>The plasmid pGC-FU-bcl-2 carrying the correct bcl-2 gene sequence could be expressed in human ovarian GC cells, and the recombinant lentivirus GC-FU-bcl-2 was generated by the packaging 293T cells. Stable expression of EGFP and bcl-2 proteins were detected by fluorescent microscope and Western blotting in 293T and human ovarian GCs after the infection. The recombinant lentivirus efficiently delivered bcl-2 gene into human ovarian GCs, in which bcl-2 expression was expressed efficiently and stably.</p><p><b>CONCLUSION</b>The recombinant lentivirus GC-FU-bcl-2 has been successfully constructed, which is capable of delivering the target gene bcl-2 into human ovarian GCs for its stable expression.</p>

Expression of vascular endothelial growth factor in nude mouse models of human endometriosis / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of vascular endothelial growth factor (VEGF) in eutopic and ectopic endometrium of in nude mice models of endometriosis and understand the role of VEGF in the development of endometriosis.</p><p><b>METHODS</b>Nude mouse models of endometriosis were established by subcutaneous implantation and abdominal injection of human endometrium xenograft from patients with endometriosis. The morphology of the foci was observed microscopically, and VEGF expression was measured with immunohistochemistry.</p><p><b>RESULTS</b>Models of endometriosis were successfully established in 8 of the 10 mice receiving subcutaneous xenograft implantation and in 7 of the 10 mice with abdominal graft injection. Increased VEGF expression was observed in the ectopic endometrium in these models.</p><p><b>CONCLUSION</b>VEGF expression is increased in the ectopic endometrium of the nude mouse models, and this animal model may facilitate future studies of angiogenesis and antiangiogenesis therapy.</p>

Construction of a recombinant adenovirus carrying endostatin gene / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a recombinant adenovirus carrying human endostatin gene with AdEasy system.</p><p><b>METHODS</b>Endostatin gene fragment was amplified from Pshuttle-Endostatin plasmid with PCR and subcloned into the pAdTrack-CMV shuttle vector. The resultant plasimid was cotransduced into E.coli BJ 5183 cells with pAdEasy-1 plasmid for homologous recombination. The linearized recombinant plasmid was subsequently transfected into AAV 293 cells, and the recombinant adenovirus was detected by GFP, PCR and restriction analysis.</p><p><b>RESULTS AND CONCLUSION</b>The positive clones of the recombinants were verified by restriction analysis and the titer of the virus reached 2.06 x 10(10)pfu/ml, suggesting successful construction of recombinant adenovirus pAd-Endo.</p>

Green fluorescent protein gene transfection mediated by recombinant adenovirus vector to rat bone marrow mesenchymal stem cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the efficient transfection of green fluorescent protein gene (GFP) mediated by recombinant adenovirus vector(Ad-GFP) to rat bone marrow mesenchymal stem cells (MSCs) in vitro.</p><p><b>METHODS</b>Wistar rat bone marrow-derived MSCs were separated and purified in vitro by Percoll density gradient centrifugation combined with adherent cell culture followed by identification with flow cytometry. MSCs infected by Ad-GFP were observed and the transfection efficiency was assessed by fluorescence microscope. The proliferative ability of these cells was tested by CCK-8.</p><p><b>RESULTS</b>The transfection efficiency was as high as 90.0%. Expression of GFP gene of infected MSCs was stable for 1 month after infection. There was no statistically difference in proliferative ability between the infected MSCs and non-infected ones (P>0.05).</p><p><b>CONCLUSION</b>The infected MSCs with Ad-GFP expressed GFP with high efficiency and retain the ability of proliferation as non-infected MSCs. Transgection with Ad-GFP is a highly effective method for labeling MSCs.</p>

Expression of hypoxia-inducible factor-1alpha in endometriosis / 南方医科大学学报

<p><b>OBJECTIVE</b>To detect the expression of hypoxia-inducible factor-1alpha(HIF-1alpha) in endometriosis and explore the possible role of HIF-1alpha in the pathogenesis of endometriosis.</p><p><b>METHODS</b>Immunohistochemistry was performed to examine the expression of HIF-1alpha in 20 normal endometrium, 20 ectopic endometrium and 68 eutopic endometrium specimens from 68 endometriosis patients, and the results were analyzed statistically.</p><p><b>RESULTS</b>The expression of HIF-1alpha was significantly increased in ectopic endometrium than in normal endometrium (P<0.01), and the expression did not undergo changes with the normal menstrual cycle in the three types of endometrium.</p><p><b>CONCLUSION</b>HIF-1alpha expression increases in ectopic endometrium, suggesting that HIF-1alpha plays an important role in the pathogenesis of endometriosis.</p>

Effect of gene polymorphism of TNF-beta on the concentration of TNF in serum of patient with endometriosis / 中南大学学报(医学版)

OBJECTIVE@#To determine the polymorphism in +252 site of tumor necrosis factor-beta(TNF-beta) gene in patients with or without endometriosis, to evaluate the levels of TNF-alpha and TNF-beta in the serum with or without endometriosis, to explore the relation between polymorphism of TNF-beta gene and the genetic susceptibility of endometriosis, and to explore the pathogenic mechanism of endometriosis at gene level.@*METHODS@#By polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method, polymorphism on +252 site of TNF-beta gene was measured in 82 patients with endometriosis (the endometriosis group) and 80 patients without endometriosis (the control group). With the sandwich-enzyme-linked immunosorbent assay (ELISA), the levels of TNF-alpha and TNF-beta in the serum of the two groups were determined.@*RESULTS@#The TNF-beta level in the serum in the endometriosis group with TNF-beta gene +252 site AA genotype significantly increased, compared with GG genotype (t=2.029, P<0.05); while TNF-alpha and TNF-beta level in the serum had no statistical significance in patients with other genotypes in TNF-beta gene +252 site in the endometriosis group and the control group.@*CONCLUSION@#TNF-beta gene +252 site AA genotype might be enhance TNF-beta level in the serum of patients with endometriosis.

Recombinant human endostatin induces apoptosis of human umbilical venous endothelial cells / 南方医科大学学报

To investigate the effects of human endostatin on proliferation of human umbilical venous endothelial cells, the apoptosis of the endothelial cells treated with recombinant human endostatin was examined by fluorescence method and the cell proliferation activity analyzed by MTT assay. The results showed that human endostatin inhibited the proliferation of the endothelial cells with ED(50) of 550 ng/ml, and the inhibitory effect was enhanced with the increase of endostatin concentration, suggesting a dose-dependent inhibitory effect of human endostatin on endothelial cell proliferation. The mechanism of antiangiogenesis induced by human endostatin is related to the inhibition of cell proliferation and induction of apoptosis of the endothelial cells.

Association of tumor necrosis factors-beta gene polymorphism with endometriosis in women in Guangdong Province / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the polymorphism of +252 site in intron 1 of tumor necrosis factor (TNF)-beta gene in relation to genetic susceptibility of endometriosis.</p><p><b>METHODS</b>Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was employed to detect the polymorphism of +252 site in intron 1 of TNF-beta gene in 82 Chinese Han patients with endometriosis in Guangdong Province and 80 Han patients without endometriosis (control group), and the relation between TNF gene polymorphism and the risk of endometriosis was analyzed.</p><p><b>RESULTS</b>The +252 site of TNF-beta allele and genotype distribution showed significant difference between endometriosis and control groups (Chi2=6.562, P<0.05; chi2=6.562, P<0.05), and relative risk of endometriosis in relation to allele A was increased by 1.793 fold. The risk of endometriosis was 3.33-fold higher in women of AA genotype than those of GG genotype (Chi2=6.562, P<0.05).</p><p><b>CONCLUSIONS</b>Allele A in TNF-beta gene +252 site can significantly increase the relative risk of endometriosis in women in Guangdong, among which TNF-beta AA genotype might be one of the genetic susceptible factors for endometriosis.</p>

Construction of a recombinant adenoviral vector expressing human endostatin / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct the recombinant adenovirus vector expressing human endostatin.</p><p><b>METHODS</b>Human endostatin gene extracted from pGEM-T Easy vector containing the target gene fragment was successfully amplified using PCR and cloned into pShuttle2 vector. The target gene was subcloned into an adenovirus vector and the resulted recombinant adenovirus (Ad-hEndo) was linearized before transfected into HEK 293 packaging cells. The Ad-hEndo recombinant adenovirus was efficiently amplified in 293T cells and purified by CsCl density centrifugation, and the titer of the virus was determined.</p><p><b>RESULTS</b>The amplified hEndostatin cDNA was verified by PCR and sequencing, and the resulted virus titer reached 5.2 x 10(9) pfu/ml.</p><p><b>CONCLUSION</b>The recombinant adenovirus containing human endostatin gene has been successfully constructed, which may provide important basis for gene therapy research for angiogenesis-dependent diseases.</p>