Diabetes mellitus (DM ) was the third largest chronic disease in the world and the number of patients in China ranked first. Diabetic nephropathy (DN ) as one of the most serious complications of DM was the main cause of death in DM patients along with the incidence of DM gradually increasing. The complex pathogenic mechanism of DN was composed of many factors such as the disorder of glucose metabolism ,inflammation and immune response. Under the special environment of the DM , autoimmunity was an important factor for the occurrence and development of DN. Immune cells ,cytokines and autoantibodies influenced each other resulting in impairment of renal function and affected the progression of DN. Further research on the immune related factors in DN and DM has important value for the prevention and treatment of these diseases.

Objective To investigate the effect of one day method in endoscopic sphincterotomy (EST) combined with laparoscopic cholecystectomy (LC) in treatment of cholecysto lithiasis and combined.Methods A retrospective analysis was carried out for fivety-seven cases of cholecystolithiasis combined with choledocholithiasis were peformed EST combined with LC as one day method (Conducted EST firstly and then took LC within 24 hours) from September 2007 to September 2012.Results Fifty-six cases completed LC,the surgical success rate was 98.24％,transit operation rate was 1.76％.Two cases performed abdominal pain and serum amylase rised the other 1 case was transited operation because of the inflammation of triangular parts of the gallbladder and discharged after 9 days.All patients total bilirubin fell to normal level within 72 hours.Conclusion The one day method in EST combined with LC treatment of cholecystolithiasis and choledocholithiasis can effectively reduce patients trauma,achieve good effect with fewer complications and shorter recovery time.

Objective To explore the clinical effects of high frequency oscillatory ventilation (HFOV) combined with pulmonary surfactant (PS) on neonatal meconium aspiration syndrome (MAS).Methods Data of 53 newborns with MAS admitted into the Neonatal Intensive Care Unit of Ning Bo Women and Children's Hospital from June 2008 to June 2011 were retrospectively analyzed.According to different therapeutic measures,they were divided into three groups:conventional mechanical ventilation (CMV) group (n=23),HFOV group (n=18) and HFOV+PS group (n=12).The oxygen index,arterial oxygen/alveolar oxygen ratio (a/ApO2) and inspired oxygen fraction (FiO2) were monitored at 2,12,24 and 48 h after mechanical ventilation.The mechanical ventilation time,duration of hospital stay,change of symptom,complications and clinical outcomes of the three groups were compared by analysis of variance and Chi-square test.Results The parameters of the three groups at 2 and 48 h after mechanical ventilation were as followed:CMV group [oxygen index:(23.79±7.27) and (15.04±4.76) mm Hg; a/ApO2:0.11±0.04 and 0.31 ±0.07; FiO2:0.74±0.16 and 0.47± 0.21],HFOV group [oxygen index:(21.13±6.29) and (11.73±4.54) mm Hg; a/ApO2:0.14±0.06 and 0.35±0.06; FiO2:0.68±0.14 and 0.41±0.11] and HFOV+ PS group [oxygen index:( 18.35 ± 5.68 ) and ( 7.85 ± 5.06 )mm Hg; a/ApO2:0.17±0.03 and 0.40±0.02; FiO2:0.59±0.13 and 0.29±0.16].Compared with CMV group,the parameters of HFOV group and HFOV+ PS group were different at different time points,and the parameters (duration and extent) of HFOV+ PS group were better than those of HFOV group (all P＜0.05).The mechanical ventilation time was (7.2±0.6) days in CMV group,(4.2± 1.4) days in HFOV group and (2.9±0.5) days in HFOV+PS group; the hospital stay was (22.2±4.5) days in CMV group,(15.6±3.4) days in HFOV group and (11.8±4.3) days in HFOV+PS group; and the oxygen treatment time was (15.4± 2.4) days in CMV group,(11.8±5.3) days in HFOV group and (7.4±2.2) days in HFOV+PS group.The mechanical ventilation time,oxygen treatment time and hospital stay time were the longest in CMV group,the shortest in HFOV+ PS group (P＜ 0.05,respectively).Conclusions Early HFOV combined with PS might be a better therapeutic method for infants with MAS than HFOV or CMV alone.

Objective To provide reference for expanding the employment positions of graduates in medical infor-mation management and information system of Chongqing Medical University by analyzing their employment situa-tion and trends.Methods The jobs of 171 graduates from Chongqing Medical University Medical Information School in 2008-2013 were registered according to the agreements of graduates, Chongqing Medical University Medical In-formation School, and employing units.Results The employment rate of graduates in medical information manage-ment and information system was relatively high .Their main employing units were medical and health organizations and associated enterprises in the City of Chongqing and Sichuan Province.The employment rate was low in coastal regions.The number of employed graduates in medical and health organizations reached its peak in 2013, and re-mained unchanged in medical and health associated enterprises.Conclusion Although the employment situation of graduates in medical information management and information system is good., the number of employing units and regions is relatively less.The literacy and ability of graduates in medical information management and information system should be improved, their employment channels should be expanded, and their professional education should be further perfected with gradual saturation of demand for graduates in employing units.

The goal of this work was to explore the prospect of standardized application of an in-vitro bioactivity assay for recombinant human follicle-stimulating hormone based on a reporter gene. The relative accuracy, intermediate precision, linearity and applicable range of the method were validated according to the General Rules of Chinese Pharmacopoeia 2020 edition Volume IV (9401). Three laboratories used this method to determine the in-vitro biological activities of six batches of drug product and three batches of drug substance manufactured by two different companies. The consistency of the potency determined by three laboratories, the intra-laboratory precision and inter-laboratory precision were analyzed. The method was optimized during the collaborative validation. The results of method validation meet the requirements of the General Rules of Chinese Pharmacopoeia 2020 edition Volume IV (9401). Aiming to resolve the problems found in the collaborative validation, the medium for cell seeding, the pre-diluted buffer solution of standard and sample, and the means of removing and discarding supernatant after stimulation were optimized. After optimization, there was no significant difference in the bioactivity among the different laboratories (P > 0.05), indicating statistical equivalency. Intra-laboratory and inter-laboratory precision were good and the geometric coefficient of variation (GCV%) were both less than 15%. In conclusion, the reporter gene assay has good intra-laboratory repeatability and inter-laboratory reproducibility and is suitable for analyzing recombinant human follicle-stimulating hormone drug product and drug substance by different manufacturers. It is expected to be used as a standardized method for the determination of the in-vitro bioactivity of such products.

BACKGROUND:Few reports are found about the effect of ovariectomized rats' bone histomorphometry parameters using non-destructive dynamic loading system.OBJECTIVE:To observe the effect of different loads situations on the bone histomorphometry parameters in ovariectomized rats.DESIGN,TIME AND SETTING:A controlled observation on the bone histomorphometry was performed in the Biomedical Engineering Laboratory of Jilin University from April 2007 to August 2007.MATERIALS:A total of 35 9-month-old Sprague-Dawley female rats were randomly divided into five groups,including sham operation group,ovariectomized (OVX) control group,OVX loading 1 N group,OVX loading 2 N group,OVX loading 4 N group.There were 7 rats in each group.METHODS:Rats in OVX control group and castration load group were processed into bilateral OVX on the back. The sham operation group only underwent the excision of fat tissues on back,and then sutured. After castration for 1 week,rats were loaded with non-destructive dynamic loading system in the two sides of the tibia,15 minutes a day. The mechanical loads would continue for 4 weeks and the loads were 1N,2N and 4N.MAIN OUTCOME MEASURES:Changes of proximal tibia bone histomorphometry parameters.RESULTS:The area,number and thickness of trabecular bone in OVX loading group were all higher than OVX control group (P＜0.05,P＜0.01).The trabecular bone area and thickness in OVX 4 N and OVX 2 N groups were significantly higher than OVX control group (P ＜ 0.001).There was a downward trend of trabecular separation in OVX 4 N group compared with OVX control group (P ＜ 0.05). With the increasing loads,there was an increasing trend of the area,number and thickness of trabecular bone,which were close to sham-operated group. The trabecular separation was declined. Single fluorescent labeled surface and double fluorescent labeled surfaces in sham operated group were all lower than that in OVX control group. With the increase in loads,the single fluorescent labeled surface,double fluorescent labeled surface,interlabeled width and mineral apposition rate had been shown to increase. The OVX 2 N and OVX 4 N groups exhibited a remarkably higher level of mineral apposition rate than OVX control group (P ＜ 0.05).CONCLUSION:With the increase in load at the range of 1-4 N,all parameters of bone histomorphometry improve in the OVX rats,the bone microstrcture is greatly ameliorated,bone mass loss is reduced and the process of osteoporosis is delayed.

BACKGROUND: The in vitro amplification is a primary method for harvesting endothelial progenitor cells (EPCs) due to its simple operation and low cost.OBJECTIVE: To isolate EPCs from rabbit bone marrow to further observe the effects of autologous EPCs on promoting vascular endothelial repair.DESIGN, TIME AND SETTING: An open experiment was performed at the laboratory of Department of Internal Medicine, Changzheng Hospital, Second Military Medical University of Chinese PLA between March 2005 and February 2006. MATERIALS: Eight New Zealand rabbits of either gender, aged 6-8 months, weighing (2.5:L-0.5) kg, were included in this study. Rabbit bone marrow was taken for isolation of bone marrow mononuclear cells by density centrifugation. METHODS: Bone marrow-derived mononuclear cells were inoculated at 1×106/cm2 and cultured for 7 days in M199 medium containing vascular endothelial growth factors and basic fibroblast growth factors. EPCs were identified by Dil-labeled acetylated low-density lipoprotein (Dil- Ac-LDL) and FITC-labeled lectin BS-1 staining. Cells that phagocytized Ac-LDL displayed red fluorescence, cells that combined with lectin BS-1 showed green fluorescence, and cells that were labeled with both exhibited orange fluorescence. Expression levels of CD133, CD134, and Flk-lwere detected using immunofluorescent staining and through the use of flow cytometer.MAIN OUTCOME MEASURES: ① Cellular morphology observation. ② Proliferative capacity of EPCs.③EPCs identified by Dil- Ac-LDL and FITC-labeled lectin BS-1. ④ lmmanohistocbemical identification of EPCs. ⑤Flow cytometry identification of EPC surface marker.RESULTS: ① Cellular morphological observation: the newly isolated bone marrow-derived mononuclear cells exhibited a round appearance. Following 72-hour culture, adherent cells grew in colony cluster, presenting with round or irregular appearance, and nuclear division was obvious. By day 7, flaky cell colonies mutually connected together, presenting with shuttle-shaped endothelioid cells.② Proliferative capability of EPCs: in the 2-4 days of culture, EPCs proliferated fast, and the proliferation slowed down thereafter, exhibiting a typical "S" -shaped appearance. By days 6 and 7, EPC proliferation accelerated again, with the absorbance values of 0.58±0.15 and 0.62±0.23, respectively. ③ Over 95% of EPC cytoplasm exhibited red fluorescence after stained with Ac-LDL, appropriately 100% of cytoplasm exhibited green fluorescence after stained with FITC-labeled lectin BS-1, and over 90% of cytoplasm exhibited orange fluorescence after double staining. ④ Immonohistochemistry and flow cytometry results revealed positive expression of EPC surface markers CD133, FIK-1, and CD34.CONCLUSION: Cell population with EPC characteristics can be successfully isolated from rabbit bone marrow by in vitro amplification.

Objective Design and synthesize short hairpin RNA (shRNA) expression vector of RNA for specific silencing of heparanase (HPA) gene,screened plasmid which silence effects is the best.Observe the function of ceil invasion after inhibiting the expression of HPA in cervical carcinoma cell lines (HeLa).Methods The genomic sequence of HPA gene was retrieved from GenBank database.Designed four pairs of specific oligonucleotide sequences and a negative control according to the shRNA design principles.They were inserted into the vector pYr-1.1,vectors,and transfected into HeLa cells via lipofectamine.Reverse transcription (RT)-PCR and immunofluorescence were employed to detect the expression of HPA gene in the transfected cells at the mRNA and protein levels,respectively.The plasmid were screened and transfected into HeLa cells,then transwell small room stromal invasion experiment were employed to observe the cervical carcinoma cell invasion.Results RT-PCR results of transfected HeLa cells shown that the mRNA amplification multiples were 0.54 ±0.05 in the HPA-592 group,0.89 ±0.18 in HPA-995 group,0.82 ±0.22 in the HPA-1351 group,0.91 ±0.47 in HPA-1658 group.While,they were 1.31 ±0.72 and 1.09 ±0.16 in negative control and blank control group,respectively.Green fluorescence was visible in the cytoplasm,which indicated that the HPA protein was expressed in the cytoplasm,of them the weakest green fluorescence in the HPA-592 group.The relative numbers of invasive cells among the HeLa cells were as follows:182 ±6 in the blank control group,258 ± 17 in the negative control group,and 44 ± 4 in the HPA-592-specific interference group(P ＜ 0.01).Conclusion Successfully screened shRNA vector targeting human HPA,efficiently inhibit expression of HPA gene when transfected into HeLa cells,and significantly reduced the invasion capacity of cervical carcinoma cells.

Objective To investigate the episodic memory monitoring ability in patients with Parkinson' s disease (PD) and explore the mechanism of the episodic memory impairment.Method The feeling-of-knowing (FOK) paradigm were established and subsequently administered in 25 PD patients and 25 healthy control (HC) participants who were matched in age and educational level.Results Compared with healthy control group ( FOK-EM recall 39.67％ ±6.11％ ; recognition 58.42％ ±7.50％ ; FOK accuracy 0.61 ±0.22),the episodic memory and its monitoring ability in PD patients were significantly impaired on the accuracy rate of FOK-EM recall ( 19.33％ ±5.10％,t =-4.833,P ＜0.01 ),recognition (45.93％ ±7.82％,t =-2.497,P ＜0.05) and FOK accuracy( -0.18 ±0.46,t =-5.986,P ＜0.01).Furthermore,the correct judgment and correct recognition of FOK-EM ( 20.47％ ± 10.78％ ) and the correct judgment and false recognition of FOK-EM (29.53％ ±5.62％ ) in the PD group were significantly higher than the HC group ( the correct judgment and correct recognition of FOK-EM:39.47％ ± 9.47％ and the correct judgment and false recognition of FOK-EM:13.90％ ±5.50％ ; t =3.564,P ＜0.05 ; t =2.306,P ＜0.05).Most importantly,the stroop effect was positively correlated with the correct judgment and false recognition of FOK-EM in PD group ( r =0.640,P ＜ 0.05 ).Conclusions In the present study,the PD patients demonstrated an overestimation of their recognition ability of episodic memory,moreover,this impairment of memory monitoring was positively correlated with the deficit of executive function,indicating that this mechanism could be an influential factor of memory disorder in PD.

Objective To detect the expression of heparanase (HPA) in cervical cancer cells and investigate the impact of quercetin on the expression of HPA,and the molecular mechanism that quercetin inhibits the growth of cervical cancer cells.Methods The experimental groups included cervical cancer cell lines (HeLa and Caski) exposed to different concentrations of quercetin (20,40 and 80 μmol/L) in the culture medium.The control groups included a negative control group,which was cultured with RPMI 1640 only,and a positive control group,in which cervical cancer cells were transfected with HPA small interference RNA (siRNA) to silence HPA expression.The cellular expression levels of HPA were detected with fluorescence quantitative real-time PCR and western blot analysis at 24,48 and 72 hours after treatment.Results (1) HPA was significantly expressed in both cervical cancer cell lines (HeLa and Caski),and it exists both nucleus and cytoplasm.(2)The real-time PCR shows as follows:as the quercetin concentration increased (20,40 and 80 μmol/L),the mRNA expression level of HPA decreased (P ＜0.01),in which the inhibition of HPA expression was concentration dependent.In addition,the inhibition of HPA expression was also time dependent.As time growth,the expression level of HPA mRNA (24,48 and 72 hours) in HeLa and Caski cells decreased (P ＜ 0.01).Compared with negative control group,the expression level of HPA mRNA decreased in different concentrations of quercetin (40 and 80 μmol/L) in both HeLa and Caski cells (all P ＜ 0.05) ; Compared with positive control group,the expression level of HPA mRNA expressed no obvious difference in quercetin (80 μmol/L) group (P ＞ 0.05) in HeLa cells,while it was opposite in Caski cells(P ＜0.01).(3)The result of western blot shown that,as the quercetin concentration increased(20,40 and 80 μmol/L)and time growth (24,48 and 72 hours),the expression level of HPA protein decreased (P ＜ 0.01),and the inhibition of HPA protein expression was concentration and time dependent.Compared with negative control group,the expression level of HPA protein decreased in different concentrations of quercetin (40 and 80 μmol/L) in both HeLa and Caski cells (all P ＜ 0.05) ;Conpared with positive control group,the expression level of HPA protein expressed no obvious difference in quercetin (80 μmol/L) group (all P ＞ 0.05) in both HeLa cells and Caski cells (all P＞0.05).Conclusion Quercetin could inhibits the expression of HPA in cervical carcinoma cell lines,which inhibition is concentration and time dependent.