Objective To establish a time-resolved immunofluorometric assay (TRFIA) to detect seurm CⅣ(collagenⅣ). Methods The antibodies to CⅣwere coated on mircoplate and the europium-labeled monoclonal antibody of CⅣ. The luminescent enhancement system was used as enhancement solution which contained mainly 2-naphthoy trifluoroacetone. we established A sandwich time-resolved fluoroimmunoassay (TRFIA) was established to measure the seurm CⅣin 127 patients with hepatitis and 30 normal controls. Results The sensitivity of assay was 12. 8?g/L. The coefficient of variation for inner-batch and inter-batch were 4. 54% and 8. 06%,respectively. The recovery was 98. 6%. The serum level of CⅣwas 46. 06?22. 21?g/L in normal control,47. 25?22. 58?g/L in acute hepatitis, 129.01?53.68?g/L in mild chronic hepatitis,277. 90?92.36?g/L in moderate chronic hepatitis,413.90?162.24?g/L in serious chronic hepatitis,568. 60?210.40?g/L in liver cirrhosis. As compared to normal control,higher concentrations of CIV (P

BACKGROUND: It has been found that vascular endothelial growth factor can induce the differentiation of bone marrow mesenchymal stem cells into endothelial cells, but can the vascular endothelial growth factor gene promote the differentiation of bone marrow mesenchymal stem cells into vascular endothelial cells in the damaged organ under the hypoxic environment? OBJECTIVE: To observe whether human bone marrow mesenchymal stem cells induced by vascular endothelial growth factor could differentiate into vascular endothelial cells under hypoxia. METHODS: The third passage of human bone marrow mesenchymal stem cells were cultured in vitro. Cells in the control group were cultured with conventional culture medium, while those in experimental group were cultured with adenovirus vector containing vascular endothelial growth factor in 5% O2. After 2 weeks of culture, morphological observation and surface-related molecular detection were performed. The levels of vascular endothelial growth factor and endothelial nitric oxide synthase were detected by ELISA. The expression of endothelin and prostacyclin was detected by RT-PCR and western blot assay. RESULTS AND CONCLUSION: (1) The number of cells in the control group was more than that in the experimental group. The cells in the control group were crowded and arranged irregularly, showing a fiber-like growth, while those in the experimental group were mostly triangular or polygonal, exhibiting a colony-like growth. (2) CD31 was negative in the control group, while CD105 was positive and the positive rate was 99.7%, indicating that the cells still showed the phenotype of bone marrow mesenchymal stem cells. The positive rate ofCD31 was significantly increased to 30.33% in the experimental group and the positive rate of CD105 expression was decreased to 58.11%, indicating a typical phenotype of endothelial cells. (3) Compared with the control group, the expression of endothelin, vascular endothelial growth factor and endothelial nitric oxide synthase increased significantly in the experimental group (P < 0.05), and the expression of prostacyclin decreased significantly (P < 0.05). All these findings indicate that vascular endothelial growth factor can promote the differentiation of human bone marrow mesenchymal stem cells into vascular endothelial cells under hypoxia.

Objective To investigate the application of hysteroscopy combined ultrasonography and laparoscopy in the diagnose and treatment of post-cesarean section scar diverticulum (PCSD). Methods From March 2011 to February 2013, 27 patients with PCSD were diagnosed and treated in Beijing Obstetrics and Gynecology Hospital. All patients were diagnosed by hysteroscopy and ultrasonography. Treatment protocols were decided by the thickness of the diverticulum. The clinical data of all patients were analysed fully. Results (1)All patients were diagnosed by hysteroscopy combined ultrasound, of which only 17 cases were diagnosed by preoperative ultrasound, the coincidence rate was 63%(17/27). (2)The thickness of diverticulum was measured by hysteroscopy combined ultrasound. Thickness of less than 3 mm in 19 cases, were treated by hysteroscopy combined with laparoscopy diverticulectomy repair;no less than 3 mm in 8 cases, were used hysteroscopy diverticulum incision. (3)The effective rate was 7/8 after hysteroscopy, and which was 16/19 after laparoscopy combined with hysteroscopy surgery. There was no difference in the treatment effect (P=0.663). The ineffective rate was 1/8 after hysteroscopy, and which was 3/19 after laparoscopy combined with hysteroscopy surgery. The size of the PCSD was smaller and the thickness of diverticulum was thicker than preoperation in the later one, which has no difference in the former one. Conclusions (1)Hysteroscopy combined ultrasound is an accurate method for the diagnose of PCSD. (2)According to the thickness of the diverticulum operation method is choosed, which is beneficial to reduce the trauma, and relieve symptoms. (3)Diverticulum poor positioning and incomplete resection are the main causes of postoperative recurrence of the diverticulum.

Objective To investigate the genetic relation between Cryptococcus neoformans var.the clinical strains in MLMT - 13 genotype and the environmental strains in MLMT - 36 genotype. Methods Multilocus microsatellite typing (MLMT) method was applied for the genotype analysis in our study.Through this method, we recognized two genotypes that distinguish a majority of clinical and environmental strains. In order to compare virulence between the two types, we chose to infect BALB/c mice (6 weeks,female) with 9 MLMT-13 strains and 10 MLMT-36 strains intravenously. Results Forty( 17 clinical and 23 environmental isolates) were analyzed. Of 17 clinical strains, 9 belonged to a major type of MLMT-13 (52.9%). They were mainly isolated from clinical specimens. About 43.5% of strains from the environment belong to a major type of MLMT-36, which are indigenous to environments and which were not isolated from clinical samples. The mortality rate and pathological changes of the above mice were observed during two months after injection. The results showed that the mortality rate of mice infected with MLMT-13 strains was 100%, while the mortality rate with MLMT-36 strains was 7. 5%. The pathological sections showed that lesions of MLMT-13 infected mice appeared in the brain, lungs, liver and kidneys, while the lesions of MLMT-36 infected mice only appeared in the brain. Most brains of MLMT-13 infected mice were distorted,and both the number and size of lesions in such brains were much larger than those of MLMT-36 infected mice. Conclusion Our study illustrated the virulent difference between MLMT-13 and MLMT-36, which are isolated from patients and environment respectively. The results inferred that some genetic changes, such ss microsatellite repeats, might occur between environmental and clinical isolates through their environmental adaptation progress.

In the present study,healthy adults who had been residing at the average altitudes of 6,2260,and 2800 m for 10 years were recruited,and the number of red blood cells,the levels of hemoglobin,and HbA1C were tested.The results showed that the level of HbA1C in altitude 2800 m group was higher compared with the other 2 groups(5.80％ ± 0.50％ vs 5.39％ ± 0.47％,P＜0.05 ).Therefore,this difference should be considered when diagnosis and treatment for diabetes mellitus were made in the region.

OBJECTIVETo compare the acute toxicity and content of daphnoretin among extracts of unprocessed indian string-bush root and its two different processed products, and to provide a basis for discussion of the mechanism of two processed methods.

METHODExtracts of unprocessed indian stringbush root and processed indian stringbush root with "sweat" and "artificial sweat" were prepared. The mice were intragastrically administrated once with these three extracts, the mortalities of mice were observed, and the median lethal dose (LD50) of different extracts were calculated with Bliss method. The determination of daphnoretin in these three samples was performed by high performance liquid chromatography.

RESULTThe LD50 of indian stringbush root extracts, indian stringbush root processed with "sweat" and with "artificial sweat" were 46.678, 72.190, 67.953 g x kg(-1), respectively. The contents of daphnoretin in unprocessed indian stringbush root, indian stringbush root processed with "sweat" and with "artificial sweat" were 0.189%, 0.407% and 0.345%, respectively.

CONCLUSIONThe toxicity of indian stringbush root processed with both "sweat" and "artificial sweat" is lower than that of the original rude drug. But the decreasion of toxicity of processed products is not by the reduced daphoretin content.

Animals ; Chromatography, High Pressure Liquid ; Coumarins ; chemistry ; toxicity ; Female ; Lethal Dose 50 ; Male ; Mice ; Plant Extracts ; chemistry ; toxicity ; Plant Roots ; chemistry ; Toxicity Tests ; Wikstroemia ; chemistry

Sixty-one newly-diagnosed diabetic patients from high altitude area were treated individually plus metformin 1.5 g/d for 14 days. The difference of oxygen partial pressure was 3.5 mm Hg (1 mm Hg=0. 133kPa) between altitude 2 260 m and 2 780 m districts. In these patients, their serum lactic acid was increased before treatment. At altitude 2 780 m district, serum lactic acid in middle-aged and elder groups were even higher [(3.90±0.85 vs 3.65±0.70), (4. 67±0.80 vs 3.69±0.78) mmol/L, both P＜0. 05＝. After 14-day treatment, serum lactic acid in middle-aged diabetic patients remained statistically unchanged [(4.50±0.50 and 3.79±0.62 ) mmol/L, P＞0.05], while that in elder patients was increased significantly [(5.59±0.55 and 5.27±0.43 ) mmol/L,P＜0. 05]. The results suggested that metformin should not be used or be used carefully in elder diabetic patients at high altitude area.

Objective To systematically evaluate the safety and feasibility of enhanced recovery after surgery (ERAS) in perioperative management of pancreatectomy.Methods Literatures were researched using CNKI,Wanfang database,VIP database,PubMed,Cochrane Library,Embase from January 1990 to March 2018 with the key words including "快速康复外科,加速康复外科,胰腺切除术,胰十二指肠切除术,惠普而术,ERAS,enhanced recovery,fast track,pancreatic surgery,pancreatectomy,Whipple,pancreatoduodenectomy,pancreatoduodenal resection".The cohort study about ERAS in elective pancreatic surgery or pancreaticoduodenectomy were received and enrolled.The patients using ERAS in perioperative management and using traditional perioperative management were respectively allocated into the ERAS group and control group.Two reviewers independently screened literatures,extracted data and assessed the risk of bias.Count data were described as odds ratio (OR) and 95％ confidence interval (CI).Weighted Mean Difference (WMD) was used as a consolidated statistics for measurement data that were measured using the same tool,and standardized mean difference (SMD) was used as a consolidated statistics for measurement data that were measured using the different tools.The heterogeneity of the studied was analyzed using the I2 test.Results Nineteen retrospective cohort studies were enrolled in the Meta analysis,and total sample size was 3 699 patients,including 1 823 in the ERAS group and 1 876 in the control group.The results of Meta analysis showed that there were statistically significant differences in the time of postoperative nasogastric tube removal,time for postoperative solid diet intake,time of postoperative defecation recovery,incidence of postoperative overall complications,incidence of postoperative delayed gastric emptying,incidence of postoperative intra-abdominal infection,duration of postoperative hospital stay and hospital expenses between ERAS group and control group (WMD=-1.70,-3.61,-0.86,OR =0.65,0.60,0.70,WMD=-4.64,SMD=-0.48,95％CI:-2.97--0.42,-4.70--2.53,-1.01--0.71,0.52-0.81,0.45-0.80,0.54-0.91,-5.91--3.38,-0.77--0.18,P ＜ 0.05).There was no statistically significant difference in the operation time,volume of intraoperative blood loss,incidence of postoperative pancreatic fistula,incidence of postoperative wound infection,readmission rate,reoperation rate and mortality between ERAS group and control group (WMD=-9.73,-14.39,OR=0.85,0.72,1.05,0.81,0.74,95％CI:-34.24-14.78,-116.96-88.17,0.72-1.01,0.46-1.14,0.83-1.32,0.58-1.13,0.53-1.02,P＞0.05).The results of subgroup analysis showed that heterogeneity of data was from eastern and western countries.Conclusion ERAS in the perioperative management of pancreatectomy is safe and feasible,it can also promote postoperative recovery of patients and reduce incidence of complications and financial burden.

Objective:To investigate the effects of doxorubicin (Dox)-loaded tumor-derived extracellular vehicles (EVs) on cell proliferation and apoptosis of human hepatocellular carcinoma.Methods:The extracellular vesicles loaded with Adriamycin (EVs-Dox) were prepared by the method of directly co-incubation. The morphology of EVs-Dox was detected by transmission electron microphotometer. The diameter of EVs-Dox was determined by dynamic light scattering (DLS). Western blotting was utilized to detect the expression of CD63, HSP 70 and TSG 101 in the EVs-Dox. The encapsulation efficiency of EVs-Dox was calculated by tandem mass spectrometry (LC-MS/MS). The drug release experiment in vitro was utilized to simulate the drug release of drug-loaded vesicles in vivo. PKH67-labeled EVs-Dox was showed cellular uptake. After treatment with EVs-Dox, MTS assay and flow cytometry assay were conducted to investigate the effects of EVs-Dox on cell proliferation and apoptosis of PLC/PRF/5.Results:The EVs-Dox showed an elliptical double-layer membrane structure of different sizes under transmission electron microscope. The diameter of EVs-Dox was (115.9±5.2) nm.Western blotting data showed high expression of CD 63, HSP 70 and TSG 101 in the EVs-Dox. The encapsulation efficiency of EVs-Dox was 0.77%. The in vitro release experiment showed that the drug-loaded vesicles could release the drug slowly. PKH67-labeled EVs-Dox showed that carcinoma cells can uptake EVs-Dox within 16h. MTS assay showed that the cell viability rate of (54.9±3.2) % was significantly lower than that of in the Dox group [(77.7±5.4)%, P<0.05]. EVs-Dox inhibited hepatocellular carcinoma proliferation. Flow cytometry assay showed that the apoptosis rate of EVs-Dox (47.9±7.0) % was higher than that in the Dox group [(38.0±1.5)%, P<0.05]. Conclusion:EVs-Dox inhibits cell proliferation and accelerates apoptosis of hepatocellular carcinoma cells.

Objective:To investigate the inhibitory effect of lycium barbarum polysaccharide(LBP)on apoptosis of Schwann cells(SCs)and its related mechanisms.Methods:The autophagy model was prepared by starvation treatment of RSC96 cells for 12 h,and the expressions of autophagy related proteins LC3 and p62 were detected by Western Blot.Cell Counting Kit-8(CCK-8)kits were used to detect the optimal concentration of LBP.RSC96 cells were randomly divided into Control group,Starvation group and Starvation+LBP group.The expressions of autophagy associated pro-teins(LC3,p62)and myelin associated proteins(p75NTR,PMP22,S100β)were detected by Western Blot or immu-nofluorescence staining.Annexin V/PI fluorescence staining was used to detect apoptosis of the cells.The cell cycle was analyzed by flow cytometry.Western Blot analysis of phosphorylation levels of pathway proteins Erk1/2 and Akt.Results:CCK8 results showed that the viability of damaged RSC96 cells was the best when LBP was 300 μg/ml.Com-pared with Control group,LC3-Ⅱ/LC3-I levels in Starvation group were significantly increased(P＜0.05).Compared with Starvation group,the proportion of apoptotic and necrotic cells in Starvation+LBP group was significantly de-creased,and the proportion of cells in S and G2/M stages was increased.The expression levels of LC3-Ⅱ,p75NTR,PMP22 and S100β were increased,while the expression levels of autophagy substrate protein p62 were decreased.In-creased expression of pathway protein p-Erk1/2(P＜0.05),while the expression of p-Akt protein decreased slightly.Conclusion:LBP can inhibit the apoptosis of SCs and promote the expression of myelin-related proteins by enhancing autophagy,which is related to the activation of Erk1/2 and/or the inhibition of Akt.