AIM: To investigate the effects of globular adiponectin (gAd) on the expression of adiponectin receptor 1 (AdipoR1) in human umbilical vein endothelial cells (HUVECs), and on the apoptosis induced by advanced glycation end products (AGEs). METHODS: HUVECs were treated with the indicated concentrations of AGEs for 24 h or 48 h in vitro. The cells in gAd treatment group were pretreated with gAd for 24 h, and then were treated with AGEs for another 24 h or 48 h. Cell variability was quantified by MTT assay. Apoptotic cells were detected by flow cytometry with Annexin-FITC/PI double staining. AdipoR1 mRNA in the cells was determined by quantitative real time reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: The viability of HUVECs treated with AGEs decreased significantly as compared to the cells treated with HSA (control, P<0.05). Under the same condition of AGEs exposure, the viability of the cells treated with gAd was greatly higher than that of the cells without gAd treatment (P<0.01). The apoptotic rate of HUVECs was significantly elevated by AGEs treatment vs HSA treatment observed by Annexin V-FITC/PI double staining analysis with flow cytometry (P<0.05). Under the condition of AGEs stimulation, the apoptosis of HUVECs was decreased by pretreatment with gAd as compared to that of the cells without gAd treatment (P<0.01). Measured by quantitative real time PCR, AGEs decreased the expression level of AdipoR1 mRNA and gAd increased the expression of AdipoR1 mRNA contrarily (P<0.05). CONCLUSION: AGEs increase the apoptotic rate of HUVECs in a concentration dependent manner and gAd promotes the AdipoR1 mRNA expression.

Objective To construct a prokaryotic expression system of human ferritin L which could be overexpressed in E.coli, and then prepare quality control materials of rFL and evaluate the homogeneity, stability, precision and application value of the rFL.Methods RT-PCR was used to clone ferritin L gene with total RNA from peripheral blood.Ferritin L gene was inserted into plasmid pGM-T to construct subclone plasmid pGM-T/ferritin L, which was identified by sequencing.The recombinant plasmid pET-30a/ferritin L was transformed into E.coli BL21 (DE3) and efficiently expressed under IPTG induction.rFL was identified by SDS-PAGE, and its antigenicity was examined by WB.The concentration of rFL was measured by ACCESS immunoassay system.Fifteen control samples were randomly selected using random number table.The homogeneity and stability of rFL were evaluated by one-way analysis of variance (ANOVA) and a linear regression model, respectively.The uncertainties of homogeneity, stability and the precision were evaluated.Results A prokaryotic expression system of ferritin L was successfully constructed.Sequence analysis showed that the ferritin L gene inserted was identical with the one from GenBank(NM_000146.3).The PCR products were 527 bp long, in complete agreement with length of ferritin L gene.The molecular weight of rFL highly expressed in E.coli induced by IPTG was 19 000 Da.The WB analysis indicated that this rFL protein had good antigenicity.The concentration of rFL(1 000 times dilution) measured by ACCESS immunoassay system was (1115.84±38.38) ng/ml.Homogeneity evaluation of low-concentration QC sample and high-concentration QC sample of rFL showed that there were no statistical significances in the within-groups and between-groups (for high-concentration F=2.336, P>0.05 and for low-concentration F=0.730, P>0.05).A linear regression based on the stability test indicated that there was no statistically significant trend of instability in five and a half months (F=1.755, P>0.05). Precision analysis showed the within-run CV, the between-run CV, the between-day CV and the total CV of high-concentration QC sample were 2.06%, 2.12%, 0% and 2.96% respectively; the within-run CV, the between-run CV, the between-day CV and the total CV of low-concentration QC sample 2.03%, 2.08%, 0% and 2.90%, respectively.Conclusion This rFL for quality control materials with independent intellectual property rights could meet the clinical requirement of homogeneity, stability and precision, and could provide raw materials for preparation of mixed ferritin for quality control.

Objective To evaluate the effect of SYNTAX score in the emergency revascularization strategy selection in ST‐segment elevation myocardial infarction patients with multi‐vessel disease(MVD) and to analyze the patient′s prognosis and influen‐cing factors .Methods A total of 144 patients with STEMI complicating MVD verified by coronary arterial angiography in the Xin‐qiao Hospital of Third Military Medical University from August 2010 to March 2012 were collected and divided into the once com‐plete revascularization group (CR group) ,staged complete revascularization group (SR group) and incomplete revascularization group (IR group) according to different emergency PCI strategies .The basic clinical conditions and coronary arterial SYNTAX score were recorded .The occurrence rate of major adverse cardiovascular events (MACCE) during 12 months follow up period was performed the statistics .The results of coronary arterial angiography were collected .The effect of different strategy on prognosis and the risk factors affecting prognosis were analyzed .Results There was no statistically significant difference in the all‐cause mor‐tality between the IR group with the CR and SR groups (P>0 .05) ,while the cerebrovascular event ,repeat revascularization during hospitalization ,AMI and total MACCE occurrence rate had statistical difference between the IR group with the CR and SR groups (P<0 .05) .The main influence factors of MACCE were complicating diabetes mellitus ,type C lesions ,SYNTAX score ,D‐B time , myocardial infarction area ,TIMI score in PCI ,no reflow existence and cardiac shock (P<0 .05) .According to the SYNTAX score , the occurrence rate of MACCE in mild or severe lesions had no statistical difference among 3 groups(P> 0 .05) ,while which in moderate lesion had statistical difference among 3 groups(P<0 .05) ,the CR group had the highest occurrence rate of MACCE . Conclusion For the patients with STEMI complicating MVD ,the SYNTAX score can be used as the evidence for selecting reperfu‐sion strategies and applied in emergency PCI .

[Objective]To investigate the effects of doxycycline on rabbit immobilized osteoarthritis and their mechanism.[Method]Twenty-four rabbits were randomly divided into normal,model,control and treatment groups,with 6 rabbits in each group.All of the animals were immobilized the lower left limbs with plaster bandage for 4 weeks except normal group.Doxycycline was orally given to treatment group.Cartilage degeneration of the medial condyle of femur was observed at 4 weeks and the levels of collagen-Ⅱ,uPA and MMP-13 were tested.[Result]Cartilage degeneration in treatment group was significantly slighter than those in model and control groups.Data of collagen-Ⅱ,uPA and MMP-13 between model and control groups had no significant difference,but a noticeable statistical difference among other groups.The uPA activity was positively correlated with MMP-13.[Conclusion]Doxycycline inhibits the degeneration process of cartilage and improves the structure.The effects may be related to the inhibition of uPA and MMP-13.

0.990).The intra-assay and inter-assay variation of the method was 3.10 % and 4.93 %,respectively.The all-trans ratinoic acid(ATRA) up-regulated t-PA mRNA expression in a dose-dependent manner(1.25~20.00 ?mol?L~(-1),P

Objective To characterize the properties of human immunodeficiency virus-1-(HIV-1)specific cytotoxic T lymphocyte(CTL)responses,and to analyze the factors which may affect CTL responses before and after treatment with highly active antiretroviral therapy(HAART)in children with chronic HIV-1 infection.Methods A total of 24 HIV-1-infected children were studied.Circulating CD4 T cell counts and HIV-1 RNA levels were routinely measured.Peripheral HIV-1-specific CTL frequency and the specific interferon-?-producing lymphocytes were determined by tetramer staining and enzyme-linked immunospot(ELISPOT)assay,respectively.Results It was found that HIV-1-specific CTL responses could be detected in vitro in more than 80% of Chinese children with HIV-1 infection.Circulating CTL frequency and IFN-?-producing lymphocyte counts were significantly reduced in HAART suppressed HIV-1 infected children in comparison with HAART failure children and HAART na?ve children.The duration of virus suppression after HAART was found to be inversely correlated with CTL frequency.Conclusion HIV-1-specific CTL response to virus was warranted by antigenic stimulation.The present work may elucidate the immune mechanisms influencing the outcome of HIV-1 carriers with treatment of HAART.

The aim of this study was to construct antisense of ZNRD1 encoding gene, and to transfect it into SGC7901/VCR cells, to seek measures to overcome multidrug resistance of gastric cancer cells. ZNRD1 cDNA amplified by PCR was confirmed by DNA sequencing, and then inserted into the multiple cloning site of the expressing vector pcDNA3 1 + with molecular cloning technique. The recombinant vector was identified by endonuclease digestion. Antisense recombinant vector was transfected into SGC7901/VCR cells using lipofectamine. Flow cytometric analysis was used to detect adriamycin (ADM) accumulation in SGC7901/VCR cells. The results showed that a fragment was obtained by PCR, and its sequence was consistent with ZNRD1 cDNA reported in the literature. After antisense recombinant vector having been transfected into SGC7901/VCR cell, ADM concentration in such cells was increased. The above results indicated that the antisense vector ZNRD1 could enhance the intracellular accumulation of ADM in SGC7901/VCR cells, which might be of potential treatment value.

Objective To study the influence of carbon dioxide pneumoperitoneum on BIS and oxygen metabolism in patients underwent laparoscopic hysterectomy in propofol and sevoflurane anesthesia. Methods 100 cases undergoing laparoscopic hysterectomy were selected in the second hospital of Shandong university and were divided into group A and group B, 50 cases in each group. Group A were received propofol and group B were received sevoflurane anesthesia . The oxygen metabolism index in the two groups were compared before and after pneumoperitoneum. Results The PETCO2 of two groups at the time points after pneumoperitoneum were significantly higher than those before pneumoperitoneum (P<0.05). There was no significant difference of the SpO2 in two groups at the different time points during pneumoperitoneum, The HR and MAP of the group A were significantly lower than that of group B (P<0.05). There were no significant difference of the PETCO2、SpO2 between two groups. Bis in 2 groups maintained a stationary state before and after pneumoperitoneum. There was no significant difference in the period between the two groups. The CjvO2、Lac of two groups were significantly higher than that before pneumoperitoneum (all P<0.05).The CjvO2、Lac of two groups were significantly lower than that before pneumoperitoneum (all P<0.05). The CjvO2 of group A at 40 minutes after pneumoperitoneum were higher than that of group B (P<0.05), and the level of CERO2、Da-jvO2 were lower than that of group B (P<0.05). But there were no difference of the BIS、CaO2 and Lac between two groups. Conclusion Carbon dioxide pneumoperitoneum can affect the BIS and oxygen metabolism in patients underwent laparoscopic hysterectomy. The effect in propofol anesthesia was more significant than that in sevoflurane anesthesia.

Objective To discuss the safety and efficacy of retroperitoneal laparoscopic partial nephrectomy in the treatment of unilateral multifocal renal tumors.Methods The data of 12 patients with unilateral multifocal renal tumors managed with retroperitoneal laparoscopic partial nephrectomy between 2009 and 2012 were reviewed retrospectively.The 12 patients had a mean age of 42 years (28 to 62) and body mass index of 23.5 kg/m2 (18.6 to 29.3),including 4 males and 8 females.Four cases were in the left side,7 cases were in the right side,and 1 case had bilateral renal tumors.The diameter of renal tumors ranged from 0.5 cm to 4.7 cm,and all tumors were projected on the surface of kidney.Nine cases were diagnosed as renal cell carcinoma,and 3 cases were diagnosed as angiomyolipoma before operation.All patients underwent retroperitoneal laparoscopic partial nephrectomy under general anesthesia,while 1 patient with bilateral renal tumors experienced radical nephrectomy of the contralateral renal tumor 3 months after the first operation.Results All the oprations were successful without conversion to open surgery.The mean renal artery cut off time,operative time,estimated blood loss and hospital stay were 36 min (26-60),126 min (75-184),142 ml (50-300) and 5 d (3-9),respectively.No complication occurred and 1-week postoperative serum creatinine was (78.1 ±8.1)μmol/L.Histopathological study revealed 9 cases of renal cell carcinomaand 3 cases of angiomyolipoma.During the follow-up for 33 months (13-51),1 patient with yon Hippel-Lindau syndrome was operated with ipsilateral radical nephrectomy for a local recurrence while the remains had no local recurrence or metastasis.Conclusions For localized and exogenous unilateral multifocal renal tumors,retroperitoneal laparoscopic partial nephrectomy is feasible and safe.Those who have hereditary and/or malignant renal tumors should be monitored closely.

Objective To compare the safety and efficiency of retroperitoneal laparoscopic and transperitoneal laparoscopic radical nephrectomy by meta-analysis.Methods A systematic review of the literature about laparoscopic radical nephrectomy was performed,searching Medline,Embase,Cochrane library,CBM,CNKI,VIP and Wan Fang database from January 2000 to October 2012.The key words were transperitoneal,retroperitoneal,laparoscopy,radical nephrectomy.Two researchers evaluated the quality of included studies.A meta-analysis was conducted using Review Manager 5.0 software.Results Nine controlled clinical studies were concluded,including 1 306 patients (520 in retroperitoneal group and 786 in transperitoneal group).The extracted data were comparable.Meta-analysis results showed that significant difference existed in operative time and complication rate (OR =16.23,95％ CI 1.62,30.84; OR =2.44,95％CI 1.35,4.41) (P＜0.05).It seemed that the retroperitioneal laparoscopic radical nephrectomy was prior to the transperitoneal laparoscopic radical nephrectomy in those items.There was no significant difference between transperitoneal and retroperitioneal laparoscopic radical nephrectomy in incision length,estimated blood loss,stay-in hospital,conversion rate,5-year disease-free survival and 5-year overall survival (P＞ 0.05).Conclusions Retroperitoneal laparoscopic radical nephrectomy demonstrated significant lower operative time and complication rate than those in transperitoneal laparoscopic radical nephrectomy.However,there is no significant difference in efficacy.Each center can choose a modality according to their convention.