Volume 89  ·   Issue 37  ·  2009

Polycystic ovary syndrome (PCOS) is one of the most common reproductive endocrine diseases in women. It begins in adolescence and is characterized by long-term ovulatory dysfunction, hyperandrogenism and polycystic ovarian changes. Its prevalence is increasing in the world and China, about 5%-10%. In recent years, it has been found that the disease not only has reproductive axis dysfunction and reproductive dysfunction, but also has a significant increase in the incidence of metabolic syndromes such as type 2 diabetes, abnormal lipid metabolism, cardiovascular diseases, endometrial cancer and breast cancer.

Gaucher's disease, also known as Gaucher's disease, is a kind of lysosomal storage disease, which is a rare familial disorder of glucose and lipid metabolism. The disease is an autosomal recessive genetic disease, which is caused by the genetic defect of β-glucose cerebrosidase in the body caused by gene mutation. Under normal circumstances, glucose cerebrosidase in human body is hydrolyzed into glucose and N-acylsphingosine under the action of this acidic lysosomal enzyme. When this enzyme is deficient, it causes substrate metabolism disorder, and a large amount of it is deposited in the mononuclear macrophage system of various organs and causes disease.

Objective To search for the role of impaired insulin signaling in the ovary reproductive failure and abnormal metabolic profiles in the AKT2 - mouse.Methods Adult,female129/C57BL/6(AKT2-)mice were used in these studies.Littermate wide types C57BL/6J(AKT2+),as well as mutant genotypes(AKT2-).The ovaries were abtained from 6 AKT2 + type mice as wall as 6 mutant genotypes,which was used for insulin stimulated glucose uptake study.By ovary transplantation on the day of 12 weeks.three genotypic mice were constructed with body AKT2 + and ovary AKT2 + in Group A,body AKT2 + and ovary AKT2- in Group B,body AKT2 - and ovary AKT2 + in Group C.The vaginal smear was done to evaluate the recovery and cyclicity of transplanted ovaries with mutant or intact AKT2.Before execution,every group was randomly separated into basal and stimulated groups in which the mice were injected recombination FSH(0.75 IU/g),and then AKT2,GSK3β,ERK-1,CYP17 and CYP19 were determined by RT-PCR in the ovaries,and the serum were reserved for the assay of HDL-C,CHO,TG,17-OHPprogesterone,E2,T,and LH.The weight of each mouse,their ovary and their fat pads and the estrus cycle,were also recorded.Results (1)The weight of fat pads beside ovaries and fold inguen in C group were significant higher than the other groups.(2)The level of 17-OHP progesterone in B group was higher than A or C group both in basal and FSH-stimulated groups.(3)In the basal group the expression of ERK-1 and CYP17 were enhanced.Moreover in FSH-dtimulated group,the expression of ERK-1,CYP17,CYP19and GSK3B in B group were higher as compared with the other groups.Conclusion (1) IR existed in the ovary of AKT2 - type,and the mice with AKT2 - type ovary had delayed cycle,PCO and high level of 17-OHP,which were similar with PCOS.(2)Metabolic dysfunction in the AKT2-mice has close relationship with whole body condition,but not defective insulin signal within ovaries.(3)Defects of insulin activity in the metabolic pathway could induce the increased expression of ERK-1 and mitogenic potential indicating the cross-talk between two pathways of insulin signaling within ovarian cells.Consequently,ovarian hyperovarianism was induced in the defective ovaries,which contribute to the enhanced response to gonadotropin and synthesis of steroid hormone.

Objective To investigate the correlations between adipocytokines and insulin resistance in women with polycystic ovary syndrome.Methods Sixty women with polycystic ovary syndrome aged 19-34years old were divided into 2 groups:group A[n=36,BMI≥25 kg/m-2 or WHR(waist height ratio)＞0.85]and group B(n=24,BMI＜25 kg/m-2 and WHR≤0.85).Twenty-six healthy infertile women with amean age of 26 4-8 years old served as controls and they were named as group C(n=26,BMI＜25kg/m-2and WHR≤0.85).Anthropometric measurements,hormonal profiles and metabolic profiles were compared between three groups.Plasma leptin,CRP and free fatty acid were measured by enzyme-linked immunosorbent assay(ELISA).Adiponectin.interleukin-6(IL-6)and tumor necrosis factor-alpha(TNF-α)were measured by radioimmunoassay.Insulin resistance was estimated by homeostasis model assessment (HOMA).Results(1)Adiponectin and TNF-α levels were significantly higher in group B(17±9)mg/Land(1.0 4±0.3)mg/L respectively than in group A(10±7)mg/L and(0.8±0.2)mg/L respectively.Leptin was lower in group B(24±13)μg/L and group C than in group A(42 ±21)μg/L(all P＜0.01).IL-6 was higher in group A(173±184)ng/L and group B(184±44)ng/L than in group C(P＜0.05 and P＜0.01 respectively).No significant difference of circulating level of CRP was found between these 3groups.(2)Leptin was positively correlated with BMI,WHR and HOMA-IR(all P＜0.01)whereas adiponectin was negatively correlated with BMI.WHR and HOMA-IR(P＜0.01;P＜0.01,P＜0.05 respectively).Mulivariate regression showed that adiponectin was the most significant predictor of HOMA-IR and it explained 16.5% of variance.Conclusion The higher levels of leptin and TNF-α,the lower level of adiponectin in obese PCOS and the higher Ievel of TNF-αtand IL-6 in non-obese PCOS suggested different adipocytokines play different roles of insulin resistance in PCOs.

Objective To analyze the clinical and metabolic characteristics of Chinese women with different menstrual types.All the women were diagnosed with polycystic ovary syndrome according to the Rotterdam consensus criteria.Methods A total of 2100 patients were divided into three groups:amenorrhea，oligomenorrhea and regular menstruation.The metabolism and endocrine indices were determined and compared among three groups.Results (1)The incidences of hirsutism and PCO were obviously higher in the amenorrhea group than in the oligomenorrhea roup(P＜0.01).(2)The amenorrhea group had the highest level of serum testosterone while the oligomenorrhea group had the lowest.Moreover，the level of luteinizing hormone(LH)was higher in the amenorrhea group than in the other two groups(P＜0.01).However the ratio of LH/FSH increased significantly in the regular menstruation group (P＜0.01).(3)The value of 2 h blood glucose was the highest in the amenorrhea group(P＜0.01).Conclusion PCOS patients with diverse types of menstrual cycle show difierent clinical manifestations and metabolic and endocrine characteristics so that the choice of treatment should be individualized.

Objective To determine the effects of hyperprolactinemia(hyperPRL)upon the symptoms of patients with polycystic ovary syndrome(PCOS).Methods Age,body mass index,levels of hormone,lipid,β-cell function and family medical history of 224 women with PCOS and 30 women with hyperPRL+PCOS were compared.Results Patients with hyperPRL+PCOS were younger to develop endocrine disturbances,an increased incidence of acne(64%vs 28%respectively),a high level of androstenedione(20±7 vs 13±5)nmol/L respectively and prolactine in serum(1492±1 175 vs 367±164)mIU/L respectively;The PCOS patients were divided into the groups of hyperandrogenism PCOS and non-hyperandrogenism PCOS depending on the serum level of androgen.A higher level of T and A was found in serum in PRL-PCOS than non-hyperandrogenism patients and similar as hyperandrogenism PCOS patients.They had reduced ApoB(680±230 nmol/L vs 943±179 mmol/L respectively)and Lpa level(46±22nmol/L vs 162 ±194 mmol/L respectively)and high HOMA-IR when compared with non-hyperandrogenism PCOS;Patients' sisters with hyperPRL+PCOS had a significantly greater incidence of acne.higher rates of infertility and PCOS when compared with PCOS patients.Levels of other hormones.metabolic profiles and other family histories did not differ between patients with PCOS and hyper-PRL+PCOS.Conclusion Patients with hyperPRL+PCOS develop the endocrine disturbances at a younger age.a greater incidence rate of acne,level of prolactin and androstenedione,they have reduced ApoB and increased HOMA-IR.Patients' sisters with hyperPRL+PCOS have significantly greater incidence of acne,higher rates of infertility and PCOS as when compared with PCOS patients.

Objective To compare the multiple pregnancy rates of frozen-thawed embryos transfer (FET)or fresh embryos transfer,analyze the factors related to multiple pregnancies after IVF and study pregnancy rates with different number of embryos transferred in FET cycle.Methods A retrospective analysis was performed upon multiple pregnancies from 1235 and 1561 clinical pregnancies conceived by FET or fresh embryo transfers.Results No correlation Was found between fresh or cryopreserved embryos transfer and multiple pregnancy rates.There were significant effects of woman's age,number of embryos transferred and stage of embryos upon multiple pregnancy rates.When the same number of cleavage-stage embryos Was transferred to women with the same age,twin pregnancy rate or triplet pregnancy rate was the same between FET and fresh cycles.Triplet pregnancy rate with three embryos transferred was significantly higher than that of two embryos transferred.In women under 35 years old,the pregnancy rate with two embryos transferred reached 36.1%.Conclusion Frozen-thawed embryos or fresh embryos transfer has no effect upon the multiple pregnancy rate.Women under 35 years old can achieve acceptable pregnancy rates when two cryopreserved embryos are transferred.It is helpful to reduce the triplet pregnancy rate.

Objective To evaluate the different influences upon the correction outcomes after anterior or posterior correction and fusion with lowest end vertebra instrumentation in adolescent idiopathic thoracolumbar/lumbar scoliosis.Methods By reviewing the medical records and roentgenograms of adolescent idiopathic thoracolumbar/lumbar scoliosis patients undergoing anterior(single rod-Group A)or posterior(all pedicle screws-Group B)correction and instrumentation,the paramaters of Cobb angle of the curve,correction rate and disc angle were measured and analyzed.Results There were 18 Patients in Group A and 21 patients in Group B.The curve included 5.0 vertebrae and 5.5 vertebrae(P=0.134)respectively,and the fusion leveis were 4.7 vertebrae and 5.4 vertebrae(P=0.008)respectively.The mean pre-operative and post-operative coronal Cobb angles of the main curve in Groups A and B were 49.2°and 10.3°,42.2°and 5.0°with an average correction rate of 78.5%and 87.8% respectively.The Cobb angles were all significantly corrected(all P=0.000),and group B had a better correction result(P=0.020).At final follow-up,the coronal Cobb angles in the two groups were 21.7°and 7.7°with an average correction loss of 8.3°and 2.7°(P=0.001) respectively.The disc angles at pre-operation,post-operation and inal follow-up were 3.1°,5.6°and 7.3°in Group A and 2.3°,4.2°and 4.4°in Group B respectively.The disc angle at post-operation was larger than that at pre-operation and the difference was significant in Group A (P=0.049) while not in Group B (P=0.050).The difference of post-operative disc angle between two groups was not significant(P=0.231). At final follow-up,the disc angle was a little larger than that of post-operation without a significant difference(P=0.112,P=0.855)while Group A had a larger disc angle(P=0.026).At final follow-up,the occurrence of proximal junctional kyphosis between Groups A and B was not significant(P=0.235).Conclusion For adolescent idiopathic thoracolumbar/lumbar Scoliosis,in comaprison with the anterior approach,the posterior approach using all pedicle screws can achieve a better correction outcome,a less correction loss of coronal Cobb angle and a better disc angle with a longer fusion range.

Objective To investigate the change of sensory gating P50 in schizophrenic patients with and without homicide.Methods The auditory evoked potentials P50 were recorded from 26schizophrenic patients with homicide(Sch group),27 schizophrenic patients without homicide(non-Schgroup)and 32 normal controls(NC)using conditioning/testing paradigm presented with auditory double click stimuli bv EGI 256 dense array.And the same time,their clinical symptoms were evaluated by positive and negative symptom scale(PANSS).Results (1)Compared with NC,two Sch groups showed no significant difference in amplitude and latency of S1-P50[amplitude:NC,Sch group,non-Sch group at Fz:(2.4 ±1.6)μV,(2.5 ±1.5)μV,(3.4±2.7)μV;latency:NC,Sch group,non-Sch group at Fz:(68 ±19)ms,(67 ±20)ms,(61 ±19)ms;respectively],but a higher amplitude and delayed latency of S2-P50[amplitude:Nc,Sch group,non-Sch group at Fz:(0.8 ±0.7)μV,(2.5 ±1.6)μV,(3.3 ±2.2)μV;latency:NC,Sch group,non-Sch group at Fz:(50 ±26)ms,(75 ±19)ms and(70 ±24)ms respectively] (P＜0.01),and no significant difference in amplitude and latency of S2-P50 between two Sch groups.(2)Compared with NC,two Sch groups showed a higher S2/S1 ratio[NC,Sch group,non-Sch group at Fz:35±26,153±137,125 ±85,respectively],lower S2-S1[NC,Sch group,non Sch group at Fz:1.69 ±1.55,0.08 ±2.41 and 0.17 ±2.30,respectively]and 100(1-S2/S1)[NC,Sch group,non-Sch group at Fz:65±26,-53±137 and -25±85 respectively](P＜0.01).And there was no significant difference in S2/S1 ratio,S2-S1 and 100(1-S2/S1)between two Sch groups.(3)Two Sch groups showed no significant difference in PANSS total,P scale,N scale,and G scale[Sch group:(110 ±27),(26 ±10),(29±7),(55±12);non Sch group:(105±27),(24±8),(28±10)and(53 ±12)respectively](P＞0.05),and no significant correlation with S2/S1 ratio,S2-S1 and 100(1-S2/S1)(P＞0.05).Conclusion Sensory gating deficit exists in schizophrenic patients with and without homicide.And it can be quantified by measuring auditory evoked potential P50,but sensory gating P50 has no difference between schizophrenic patients with and without homicide.

Objective To observe the effects of estrogen depletion and 17β-estradiol replacement therapy upon ratβ-amyloid(Aβ)generation and the possible related mechanisms.Methods Rat ovaries were ectomized to mimic estrogen-depletion models and then 17β-estradiol was administered by powdering hormone into soy-free chow as a way of replacement therapy.ELISA was carried out to detect rat hippocampus Aβ levels and α-and β-secretase activities were measured after the experiment. The effects of estrogen depletion and 17β-estradiol replacement therapy upon -βsecretase(BACE1)and neprilysin(NEP)expression were also analyzed by Western blot.Results Ovariectomy significantly decreased estrogen level [(11 ±4)pg/ml,P＜0.01]as compared with control group[(21 ±8)pg/ml]while 17β-estradiol administration increased the estrogen level[(63±13)pg/ml,P＜0.01]in blood.The Aβ[(28.5±4.5)ng/ml,P＜0.01]and Aβ42[(4.5±1.2)ng/ml,P＜0.01]levels were higher in ovariectomy group as compared with their respective control group[with Aβ40(14.4±2.4)ng/ml and Aβ42(2.8 ±0.4)ng/ml respectively].But the effects of ovariectomy on Aβ content can be partially reversed by 17β-estradiol replacement therapy[with Aβ40(20.3±3.2)ng/ml,P＜0.01 and Aβ42(3.8±0.5)ng/ml,P＜0.01 respectively].Estrogen depletion decreased α-secretase activity(67.5%,P＜0.01)and increased β-secretase activity(145.8%,P＜0.01)and this effect can be blocked by 17β-estradiol administration [ with α-secretase activity to 90.2%(P＜0.01)and β-secretase activity to 92.4%(P＜0.01)].Ovariectomy increased BACE1 expression(135.4%,P＜0.01)and decreased NEP expression (40.8%,P＜0.01)and this effect can be partially antagonized by 17β-estradiol supplementary[with BACE to 103.5%(P＜0.01)and NEP to 88.4%(P＜0.01)].Conclusion Estrogen depletion can increase Aβ generation through the effects of increased β-secretase activity and decreased α-secretase activity.Ovariectomy also increases BACE1 expression and decreases NEP expression.The 17β-estradiol supplementary can decrease Aβ generation and this may to some extent explain why estrogen replacement therapy can decrease the risk of Alzheimer's disease in postmenopausal women.

Objective To evaluate the conformablity and quantization controllability of multi-cluster conformable radiofrequency ablation electrode by ex vivo animal experiment.Methods The subject using in this experiment were fresh ex vivo ox liver.The electrode we used were the series of WHK-3(including WHK-3Ⅰ and Ⅱ),which were design and made by Wellfare Electronic Technological Company(Beijing).Each electrode had 6 sub-electrode on opposite side(A and B).Both A and B side could separately in different length and radian so to form asymmetric shape.The difference between WHK-3Ⅰand Ⅱ were the shaft of Ⅰ are electric conductive while it of WHK-3Ⅱwere inconductive by coating with insulant material.Our subject were divide into 3 group.In group 1 we do radiofrequency ablation in ex vivo ox liver with both sub-electrode(WHK-3Ⅰand Ⅱ)fully spread out symmetrically,then the diameter of 3D cross section in zoue of melt necrosis between type Ⅰ and Ⅱ were compared.In group 2 we do radiofrequency ablation in ex vivo ox liver with WHK-3 Ⅰ while the two side of sub-electrode were spread out asymmetrically in different shapes.Then the diameter of Cross section which were parallel to the electrode shaft in zone of melt necrosis in difierent sub-electrode shape were compared.In group 3 we do the test same way as in group 2 on WHK3 Ⅱ electrode.Results In group 1,there had no significant different between the diameter of melt necrosis zore in ex vivo ox liver with WHK-3ⅠandⅡ(average around 6×6×3 cm3)while sub-electrodes were fully spread out symmetrically.In group 2,while sub-electrode in A side were fully spread and sub-electrode in B side were half spread,WHK-3Ⅰelectrode could form an asymmetric melt necrosis zone.The diameter on A side were 3.24±0.32 cm and it on B side were 1.87±0.24 cm.While only A side sub-electrode were spread out,the diameter on A side were nearly the same and it on B side were decreased to 1.55±0.16 cmIn group 3,while sub-electrode in A side were fully spread and sub-electrode in B side were half spread,WHK-3Ⅱelectrode could form an asymmetric melt necrosis zone too.The diameter on A side were 3.27±0.35 cm and it on B side were 1.65±0.23 cm. While only A side sub-electrode were spread out,the diameter on A side were nearly the same and it on B side were decreased even to 0.90±0.18 cm.Conclusions The series of WHK-3 could form an asymmetric melt necrosis zone in ex vivo tissue,which were more suitable to clinical usage.And both of them had quantization controllability,especially in WHK-3Ⅱ.

Objective To construct the point mutation plasmids expressing NS3/4A with different secondary structure of the amino-terminal 120 residues of NS3,and to investigate the differenees in serine protease and inhibitory effects on host cells between each subgroup.Methods The point mutation plasmids were constructed,which expressed NS3/4A with the corresponding secondary structures of subgroup,and were named as A1-2,A2-1,A2-2,B1-1,B1-2,B2-1,and B2-2,with the backbone of M-H05-5(A1-1).Western blot was performed to detect the expression of NS3/4A and the difference in in cis and in trans NS3 serine protease activity between each subgroup.The inhibitory effects of HCV NS3/4A with different aminoterminal secondary structures on IFN-β production and p53-dependent transcriptional activation were revealed by Luciferase reporter assay.Results Western blot revealed the successful expression of the constructs and the incomplete cleavage of NS3/4A in subgroup A2-1 and B2-1.indicating that the in cis NS3 serine protease activities of subgroup A2-1 and B2-1 were weaker compared with that of the other subgroups.By using NS5A/5B△C as a substrate for NS3/4A serine protease,it was also found that the in trans NS3 serine protease activities of subgroup A2-1 and B2-1 were also weaker compared with that of the other subgroups.Differences in inhibitory effects of HCV NS3 on IFN-β promoter activity and on p53-dependent lucifevase gene transcriptional activation were also observed between subgroup A2-1,B2-1 and the other subgroups.Conclusion HCV NS3/4A with different secondary structures at amino-terminus has different serine protease activities and inhibitory activities on host cell functlons.

Objective To examine the effect of TNF-related apoptosis-inducing ligand(TRAIL)and in combination with doxorubidcin (ADM) to xenografted tumors in nude mice and to explore its potential mechanism.Methods MG-63 cells(5 x 106/ml) were suspended in 0.2 ml RPMI-1640 and inoculated subcutaneously into the lower limb of nude mice.Treatment groups were given TRAIL of different concentration or combination of TRAIL and ADM intraperitoneally.Normal saline was administrated in the control group.Auti-tumor effects were estimated by tumor volumes.Serum alkaline phosphatase(ALP)was detected by ALP kits.Induction of apoptosis in xenografted tumors was confirmed by TUNEL(TdT-mediated dUTP nick end labeling)assay.Expression of Bax was detected by immunohistochemical assay.Expression of TRAIL receptors was detected by RT-PCR assay.Results Growth curve of tumors indicated that tumors carried by TRAIL-treated mice grew more slowly than that with normal saline and 2μg TRAIL was more effective,Also tumors treated with combination of TRAIL and ADM grew more slowly than any other group.ALP activities of each group were moderately different but significance was not reached.TUNEL showed that there were more apoptotic cells in the combination group than any other group.Immunohistochemical assay showed that expression of Bax was up-regulated in the combination group.RT-PCR showed that expression of TRAIL-R2 mRNA was up-regulated in the combination group.Conclusion TRAIL can induce an effective apoptosis of osteosarcoma cells in vivo in a dose-dependent fashion.ADM can enhance the effect of TRAIL-mediated apoptosis.And up-regulations of Bax and TRAIL-R2 may be the involved mechanism.

Objective To observe the change of expression of vascular endothelial growth factor (VEGF)and vascular leakage in the brain of rats exposed to high altitude with siRNA targeting Vascular endothelial growth factor and explore the pathological mechanism and preventive approach of high altitude cerebral edema(HACE).Methods Fifty male Wistar rats were divided randomly into normal control group (Ncon),hish altitude control group(Hcon),intraventricular normal saline control group(Scon),intraventricular siRNA group(CVI)and intravenous siRNA group(IVI).Rats in Ncon were raised normally.Rats in Hcon,Scon,CVI and IVI pretreated with intravenous injection of normal saline,intraventricular injection of normal saline,intraventricular injection of siRNA and intravenous injection of siRNA respectively were exposed to a low-pressure cabin mimicking a high altitude of 7000 m for 24 h.The ratio of dry and wet brain weight was calculated and the sodium fluorescein leakage calculated to evaluate the cerebral edema and the blood brain barrier permeability.Also the real-time quantitative RT-PCR was employed to detect the expression of VEGF mRNA and the Western blot the expression of VEGF.Results Compared with rats in NC,high altitude exposure led to a significant increase in the levels of VEGF mRNA (from 21.6±3.5 Kcopies/μg to 36.3±3.9 Kcopies/μg,P＜0.01)and protein(from 48±0.09 to 0.77±0.12,P＜0.01)in rat brain and fluorescence intensity of sodium fluorescein increased significantly(from548±48 rfu/mg to 674±32 rfu/mg.P＜0.01).Intravenous injection of siRNA targeting to VEGF caused no significant change of expression VEGF mRNA and protein and fluorescence intensity of sodium fluorescein in rat brain(P＞0.05,respectively).While compared with rats in HC,intraventricular injection of siRNA targeting to VEGF caused the significant reduction of expression of VEGF mRNA (from 36.3±3.9 to 19.9±4.3,P＜0.01)and protein(from 0.77±0.12 to 0.44±0.13,P＜0.01)and fluorescence intensity of sodium fluorescein(from 674±32 rfu/mg to 542±77 rfu/mg,P＜0.05)in rat brain.There were no significant change in the ratio of dry and wet brain weight among five groups.Conclusion VEGF may play a key role in the pathologic process of HACE.Intraventricular injection of siRNA targeting to VEGF inhibits the expression of VEGF and prevent the high altitude-induced vascular leakage.These findings might provide a basis for new preventive approaches of cerebral edema.

Objective To investigate the brain injury of neurocytes in hippocampus of adenosine A1 receptor knock-out mice during pentetrazole kindling and detect the correlation of brain injury and the expression of COX-2 so as to evaluate the neuroprotective function of adenosine A1 receptor and its mechanism.Methods The animals were divided into two groups:wild type group and KO group.The kindling model was established by iniection of pentetrazole into abdominal cavity.The expression of brain injury related protein,casepase-3 and COX-2 in hippocampus was investigated separately at different time points(24 hour,1 month) post-kindling.Results The expression of Caspase-3 and COX-2 increased in KO group both during acute and chronic phases post-kindling compared with normal mice.There was statistical difference(P＜0.05).Furthermore,the expression levels of two proteins were higher at 1 month compared with 24 hour(P＜0.01).These results indicated that the increased expression of Caspase-3 and COX-2 post-kindling was earlier and much more in KO mice as compared with wild-type ones.Conclusion Adenosine exerts strong neuroprotective functions through the activation of adenosine al receptor.This receptor reduces cell apoptosis during pentetrazole kindling.Cyclooxygenase-2(COX-2)reflects the extent of brain injury.The neuroprotective function mediated by adenosine A1 receptor might be related with COX-2.

Objective To understand the biological characteristics and osteogenic potential of hVEGF-165 gene modified marrow stromal stem cells and investigate the effect and value of treatment for osteonecrosis of femoral head by hVEGF-165 gene modifled marrow stromal stem cells under arthroscope.Methods rAAV-2-hVEGF-165 plasmids were extracted and transfected into rabbit marrow stromal stem cells.hVEGF-165 mediated by adeno-associated virus(AAV)was used to transfect rMSCs.The transfection efficiency was detected with enhanced green fluorescent protein under fluorescence microscope.hVEGF-165mediated by adeno-associated virus(AAV)was used to transfect rMSCs.Virus transfection stayed overnight after 90% cell converged.MOI Wag 105.The transcription and expression of hVEGF-165 protein expression were detected by RT-PCR and Western blot.The necrotic bone was emptied and then MSCs were implanted under arthroscope.The histology of femoral head was inspected at postoperative 2-8 weeks.Results The expression of hVEGF-165 gene could be found distinctly in the transfected rabbit MSCs and hVEGF-165protein in the supernatants of transfected cell cultures.The transfection efficiency of adeno-associated virus (AAV)transfected rMSCs was 70%.And rAAV-2-hVEGF-165 transfected rMSCs achieved an effective expression by RT-PCR and Western blot.hVEGF-165 could be found after a 48-hour transfection and peaked at Day 10.Immunohistochemical detection showed that the implanted rMSCs was positive at Week 2 and strong positive at Week 8.The compressive strength of the hVEGF-165 gene group approached that of normal control.Conclusion hVEGF-165 gene transfected rabbit MSCs can express hVEGF-165 with highly biological activity.It provides provided a basis for employing hVEGF-165 gene and MSCs based gene therapy for ONFH repairing and regeneration.rAAV-2-hVEGF-165/MSCs may be implanted accurately under arthroscope.Implantation of human BMP-2 gene transfected BMSCs can repair early-stage experimental femoral head necrosis.

A 32-year-old married female was admitted to the First Hospital of Zhejiang University on October 12, 2008 due to irregular vaginal bleeding for 1 month. Gynecological examination showed that a cauliflower-like mass of-3 cm ×3 cm ×2 cm was seen at 6 o'clock of the cervix, bulging towards the cervical canal and cervical surface, superficial erosion, contact hemorrhage, vaginal fornix, no obvious thickening of bilateral parauterine, normal size of uterine organs, medium texture, no obvious mass in bilateral appendages, no tenderness. Blood tumor markers (Cal25, Cal99), alpha-fetoprotein (AFP), serum human chorionic gonadotropin (HCG) and cervical squamous cell carcinoma antigen were all in the normal range.

A 12-year-old female was admitted to the hospital mainly because of "repeated sleepiness, visual decrease for 3 years and aggravation for 2 days". The patient developed fatigue more than 10 days after being vaccinated with Japanese encephalitis vaccine in May 2005, and then developed symptoms such as frequent vomiting, low fever, poor spirit, limb weakness and unstable walking. After 4 days of symptomatic treatment in the local hospital, the effect was poor (specific diagnosis and treatment is unknown). In mid-May, the above symptoms were significantly aggravated and then transferred to a hospital in Beijing. Brain MRI showed that there were multiple abnormal signals scattered in the left cerebellar hemisphere, vermis, parenchyma of bilateral cerebral hemispheres, basal ganglia and corpus callosum.