Objective To investigate the developmental profiles on surface expression and colocalization of NMDA receptor and AMPA receptor on dendritic structures in cultured hippocampal neurons of rats. Methods Two vectors expressing green fluorescent protein tagged GluR1 subunit(GFP-GluR1) and FLAG tagged NR2B subunit(FLAG-NR2B) were co-transfected into cultured hippocampal neurons at 5 days in vitro(DIV5).Surface expressed GFP-GluR1 containing AMPA receptor clusters and FLAG-NR2B containing NMDA receptor clusters were then labeled in living neurons by using anti-GFP primary antibody followed by Alex488-conjugated secondary antibody,and anti-FLAG primary antibody followed by Cy3-conjugated secondary antibody.Furthermore,distribution of the surface clusters was observed on the detailed dendritic structures visualized with co-expression of cyan fluorescent protein tagged actin(CFP-Actin). Results The numbers for GluR1 containing AMPA receptor clusters,NR2B containing NMDA receptor clusters,and colocalized receptor clusters per 100*!?m dendrite in DIV7 and DIV14 neurons were 59.2?5.6 and 74.8?3.1(P

AIM: To obtain GST fusion protein of hSP17 gene and construct the recombinant plasmid for expression in E. coli. METHODS: Total fragment of hSP17 cDNA gene were amplified by RT-PCR, then subcoloned into pGEX-3b to generate recombinant hSP17/pGEX. Right orientation of insert are identified by restricted enzyme digestion. Transform the correct recombinant plasmid into the E. coli DH5a. The expression of fusion proteins hSP17-GST were induced by adding isopropylthiogalactoside (IPTG). RESULTS and CONCLUSION: The recombinant plasmid hSP17/pGEX-3b could express effectively in E.coli and a high level of fusion protein hsp17-GST with the predicted molecular weight was detected.

Objective To establish and optimize three-cube FRET assay in living cells and analyze subunit assembling of iGluR receptors. Methods Taking HEK293 cells cotransfed with pECFP and pEYFP as negative control, and those transfected with pECFP-YFP as positive control,different calculation methods using fluorescence microscopy were compared. Results These calculation methods were all suitable for FRET measurement in the system. but the measurement results were affected by the ratio of Donor/Acceptor (D/A) in some degree,and different calculation methods have different optimized conditions. FRET measurement using FR value showed subunit specific assembly of iGluR subtypes.Conclusion There are different optimized conditions for these different calculation methods in the three-cube FRET measurement system,and a further evidence is provided for subunit specific assembling of iGluR subtypes from the FRET assay.

Objective To investigate the effects and outcomes of four procedures for hysterectomy. Method Retrospectively, we analyzed the clinical characteristics of 756 cases who respectively received hysterectomy by total abdominohysterectomy (TAH,260 cases), modified abdominohysterectomy (MAH, 180 cases), transvaginal hysterectomy (TVH, 106 cases), or laparoscopic assisted vaginal hysterectomy (LAVH)(210 cases) and compared their effects and outcomes. Results The average operation time of TAH(98 ?23)minutes, MAH(67?18)minutes, TVH (63?19)minutes and LAVH(99?35)minutes. The average operation time of LAVH and TAH groups was significantly longer than that of TVH and MAH groups (P

Objective To investigate the preventative effect of perioperative intervention to lower extremity deep venous thrombosis (DVT) after gynecologic surgery. Methods The 1062 patients, who received gynecologic surgery during 2007 June to 2008 June, were derided into intervention and nonintervention groups randomizely. According to the risk factors of DVT, the patients in intervention group were divided into 4 subgroups: low, mid, high and extremely high rise Each group had its own preventive measures. If patients had spontaneous pain, tenderness and swelling, positivity of Neuhof or Homan syndrome, and extension of one low extremity superficial vein, the low extremity color Doppler ultrasound would be carried out immediately. The ultrasound would also be routinely carried out in the high and extremely high risk groups in intervention group after 2 and 7 days after surgery. A prospective study was carried out investigating incidence of DVT and coagulation function perioperation. The vein blood samples were taken at in a week before surgery and 48 hours post-operation. Results The incidence of DVT of intervention group was 1.10% (6/546), and in nonintervention group, the incidence was 3.29% (17/ 516). There was significant difference between two groups (P<0.05). The incidence of DVT in extremely high risk subgroup was 21.05% (4/19), which was significantly higher than that of low(0), mid(0) and high risk groups[2.13% (2/94), P<0.05]. D-dimer, antithrombin-Ⅲ(AT-Ⅲ) post-operation were all higher than that pre-operation in the two groups, but there was no significant difference (P>0.05). The indexes of coagulation system, such as blood platelet count, prothrombin time (PT), active partial throraboplastin time (APTT), fibrinogen (Fbg), thrombin time (TT), tissue-plasminogen activator (t-PA), plasminogen activator inhibitor(PAI), were not significantly changed perioperation (P>0.05 in all pre-or post-operation indexes). Conclusions Perioperative intervention measurement according to different risk of DVT could cut down the incidence of DVT. DVT preventive intervention is suggested to high and extremely high risk people.

Objective:To obtain a high specificity and high affinity anti-human PD-L1 monoclonal antibody which can be used for clinical diagnosis and block PD-L1 and PD-1 binding.Methods:BALB/c mice were immunized with recombinant PD-L1 protein.The positive cell clones stably secreting anti-human PD-L1 monoclonal antibody were obtained by classical hybridoma cell fusion technique.The specificity,affinity,subtype and other characteristics of the antibody were identified by ELISA.Immunofluorescence and indirect immunofluorescence were used to detect the tumor cells.Antibody blocking activity was confirmed by tumor killing test.Results:Two cell strains stably secreting monoclonal antibodies against human PD-LI were screened out.Abl and Ab2 had high titer and affinity.The antibody titers were 1:2.56×106 and 1:3×105,and the affinity was 1.5×109 L/mol and 2.5×10s L/mol respectively.There was no cross reaction between these two antibodies and PD-L2.Immunoblotting,indirect immunofluorescence confirmed that the antibody can be used to the diagnosis.Experiment showed that PD-L1 antibodies can increases tumor-killing activity of CIK cells.Conclusion:Two hybridoma cell lines capable of stably secreting highly specific and high affinity anti-human PD-L1 monoclonal antibody are obtained.They can specifically bind to PD-L1 molecules on tumor cells and can be used to the diagnosis of tumor phenotype and prognosis.Antibody blocking function can be applied to combined CIK cell immunotherapy.

OBJECTIVETo observe the clinical effects and the serous estradiol change of patients with menopause syndrome using Xianling Gubao capsules for treatment.

METHOD110 patients with manopause syndrome who were diagnosized by clinic index were recruited for the study. They were divided into two groups randomly. The treatment group were dealed with Xianling Gubao capsules and the control group with placebo of starch capsules for 8 weeks. The menopausal Kupperman index was used to assess change in menopausal symptoms before and after treatment. At the same time, serous estradiol and the thickness of uterine endometrium was evaluated.

RESULTThe Kupperman index were improved apparently in treatment group after treatment (P < 0.01). The serous estradiol level of treatment group was increased apparently after treatment (P < 0.05). However, There were no statistical deference in the thickness of uterine endometrium in the treatment and the control group before and after treatment.

CONCLUSIONThe treatment using Xianling Gubao capsules for patients with menopause syndrome could improve the clinic symptoms, increase the serous estradiol level, but do not increase the risk of endometrial hyperplasia.

Capsules ; Drugs, Chinese Herbal ; therapeutic use ; Endometrium ; drug effects ; metabolism ; Female ; Humans ; Menopause ; drug effects ; metabolism ; Middle Aged ; Treatment Outcome

Objective Previous studies on the association between lipid profiles and chronic kidney disease(CKD)have yielded inconsistent results and no defined thresholds for blood lipids. Methods A prospective cohort study including 32,351 subjects who completed baseline and follow-up surveys over 5 years was conducted.Restricted cubic splines and Cox models were used to examine the association between the lipid profiles and CKD.A regression discontinuity design was used to determine the cutoff value of lipid profiles that was significantly associated with increased the risk of CKD. Results Over a median follow-up time of 2.2(0.5,4.2)years,648(2.00%)subjects developed CKD.The lipid profiles that were significantly and linearly related to CKD included total cholesterol(TC),triglycerides(TG),high-density lipoprotein cholesterol(HDL-C),TC/HDL-C,and TG/HDL-C,whereas low-density lipoprotein cholesterol(LDL-C)and LDL-C/HDL-C were nonlinearly correlated with CKD.TC,TG,TC/HDL-C,and TG/HDL-C showed an upward jump at the cutoff value,increasing the risk of CKD by 0.90%,1.50%,2.30%,and 1.60%,respectively,whereas HDL-C showed a downward jump at the cutoff value,reducing this risk by 1.0%.Female and participants with dyslipidemia had a higher risk of CKD,while the cutoff values for the different characteristics of the population were different. Conclusion There was a significant association between lipid profiles and CKD in a prospective cohort from Northwest China,while TG,TC/HDL-C,and TG/HDL-C showed a stronger risk association.The specific cutoff values of lipid profiles may provide a clinical reference for screening or diagnosing CKD risk.

Objective To explore the current situation and influencing factors on Classroom Mobile Phone Dependence Syndrome (CMPDS) among college students,and to provide scientific basis for guiding college students to use mobile phones reasonably and healthily.Methods Stratified cluster sampling method was used.Students from different majors and different grades in Lanzhou University were included as the research objects.Classes were recognized as a unit in receiving basic field investigation in this questionnaire related study.Informed consent principles were followed and process of survey was anonymously carried out.Results The overall rate of CMPDS in college students was 8.7％,including ‘mild rate'as 6.6％ and ‘seriously mild rate'as 2.1％.No significant differences were found on genders or grades.Factors as shopping in the classroom shopping (OR=3.720),being bored on courses (OR=1.740),WiFi coverage (OR=1.787),time of practice in the classrooms (OR=1.514),and the total time of daily mobile phone use (OR=1.513) etc,appeared as risk factors related to CMPDS among the college students.However.shooting courseware (OR=0.579) appeared as a protective factor.Conclusions Rate of CMPDS was high in college students and we suggested to form a joint task force among the college authority,teachers and students to work on the related problems.Hopefully,the serious CMPDS condition will be minimized and both physical and mental health of the college students be improved.

[Objective]To investigate the protective effect of Zhuangtongyin on the Middle Cerebral Artery Occlusion(MCAO)model by modulating ferroptosis through the Nrf2-SCL7A11/xCT-Gpx4 pathway and its underlying mechanism.[Methods]C57BL/6J mice were randomly divided into Sham operation group(Sham),model group(MCAO),low-dose Zhuangtongyin group(ZTY-L),high-dose Zhuangtongyin group(ZTY-H),with 5 mice in each group.The MCAO group was modelled by silica gel embolization,the middle cerebral artery of mice was embolized for 1h,then the silica gel was pulled out and reperfusion was performed after 72 h;and the other operations in the Sham group were the same as those in the MCAO group except that the thread plug was not inserted.The neural function of mice was evaluated by Zea-Longa method.TTC staining was used to evaluate the volume of cerebral infarction.The level of brain injury was evaluated by HE staining and Nissl staining.Prussian blue staining and the expression of iron transport-related carrier receptors TfR1 and DMT1 on mRNA level was detected by qPCR to evaluate the iron ion deposition level in mice brain.The expression of lipid peroxidation-related gene ACSL4 on mRNA level was detected by qPCR,and the content of 4-HNE was detected by ELISA kit to evaluate the lipid peroxidation level of mice brain.The expressions of ferroptosis marker PTGS2 mRNA level was detected by qPCR.The expressions of Nrf2,SCL7A11/xCT,Gpx4 in mice brain tissue were detected by Western-blot and immunofluorescence.[Results]Zhuangtongyin improved the nerve function of mice after MCAO(P＜0.05)and the cerebral infarction volume of mice(P＜0.05)and alleviate the pathological injury of cerebral cortex cells after MCAO operation.Zhuangtongyin attenuated the accumulation of trivalent iron ions in the brain tissue of mice following MCAO.Additionally,Zhuangtongyin downregulated the expression of TfR1 and DMT1 mRNA(P＜0.001),a transporter associated with cellular iron ion uptake,in the brains of post-MCAO mice.Furthermore,Zhuangtongyin reduced levels of lipid peroxidation product 4-HNE(P＜0.001)and suppressed ACSL4 mRNA expression in brain tissue post-MCAO(P＜0.001).Besides,Zhuangtongyin downregulated the expression of PTGS2 mRNA(P＜0.001),in the brains of post-MCAO mice.Zhuangtongyin increased the expression of nrf2 into the nucleus(P＜0.001),and increased the expression of xCT and Gpx4 in neurons after MCAO(P＜0.001).[Conclusion]Zhuangtongyin can enhance the nerve function and reduce cerebral infarction volume in MCAO/R mice,alleviate the pathological damage of cerebral cortex cells,and modulate the expression of key signaling molecules in the Nrf2-SCL7A11/xCT-Gpx4 pathway.Therefore,it is suggested that the mechanism by which Zhuangtongyin improves MCAO/R injury in mice may involve regulating ferroptosis through the Nrf2-SCL7A11/xCT-GPX4 pathway.