Objective:To value the application of two bacteria colony-PCR methods in the screening of phage antibody library. Methods:Five positive monoclonal bacterium were respectively suspended in either deionized water or 0.1% Triton X-100, and then boiled to be used as template in PCR. . The DNAs products of PCR were extracted and digested by two enzymes, and then determined by electrophoresis. Results:The inserted genes were detected in all the 5 clones after PCR and enzyme digestion .Conclusion:Bacteria colony-PCR can be used in screening positive recombinant colonies. The bacteria colony-PCR method with bacteria colonies suspended in deionized water is valuable in large scale positive recombinant bacterium screening.

Objective To investigate the role of autophagy and synaptophysin (SYP) in cognitive impairment in de?pressed rats receiving electroconvulsive shock (ECS). Methods Clean and healthy adult male Sprague-Dawley rats were acclimatized to a standard laboratory environment for 7 days. The chronic unpredictable mild stress (CUMS) was used to establish the rat model of depression. Behavior tests were conducted before and after CUMS to evaluate the depression and cognition level of rats. After establishment of the model, 24 rats were randomly divided into ESC group (group E) and depression group (group D) with 12 rats in each group. The rats in group E were administered 80 mg/kg of propofol (10 mg/mL) by intraperitoneal injection, followed by ECS treatment. The rats in group D were administered propofol by intra?peritoneal injection, followed by sham-ECS treatments. The above interventions were conducted daily for 7 consecutive days. After the interventions, rats underwent behavior tests as before. Subsequently, rats were killed and specimens were collected for measurements. Immunohistochemistry was performed to examine autophagy markers such as Beclin 1 and LC3Ⅱand ELISA was used to detect SYP in the hippocampus. Results Group E after ECS significantly increased the percentage of sucrose preference (68.2%±8.7%), rearing times (7.0±1.9), total horizontal distance [(569.5±70.0) cm], es? cape latency [(21.9±5.3)s] and space exploration time [(20.5±3.9)s] compared with group D or group E before ECS. There was no significant difference in these index between groups before ECS or in group E between before and after ECS(P>0.05). Compared with group D, group E had upregulated protein expression levels of Beclin 1 and LC3Ⅱin CA1, CA3, DG as well as the area near the hippocampus and increased SYP contents (P<0.05). Conclusions Cognitive impairment in depression rats following ECS correlates with activated autophagy and increased SYP by ECS.

Objective To investigate the effect of controlled low central venous pressure(CLCVP) on blood loss and prognosis in different types of hepatectomy .Methods Two hundred and fifty seven patients underwent standard hepatectomy ,half liver resec‐tion or irregular partial hepatectomy from January 2011 to December 2012 in the First Affiliated Hospital of Chongqing Medical U‐niversity were retrospectively studied .Patients treated with CLCVP during hepatectomy were attributed to the CLCVP group .CVP of these patients were lowed to below 5 cm H2 O by minimizing fluid infusion and one or both of the following maneuvers :posture adjustment ,nitroglycerin administration .Alpha agonists were used when necessary to maintain the mean arterial pressure MAP at ≥60 mm Hg .Other patients been maintained with normal level of CVP by adjusting fluid administration were included in normal CVP group (NCVP) .Blood loss and transfusion volume ,length of hospital stay of the two groups were compared ,and the effects of different surgery type on CLCVP blood protection were evaluated .Results In the patients underwent standard hepatectomy or half liver resection ,intraoperative blood loss and transfusion were not statistically different between the two groups .While in the pa‐tients underwent irregular partial hepatectomy ,the CLCVP group suffered less blood loss and transfusion(P<0 .05) .Percentage of the patients with less than 200 mL blood loss and no transfusion of concentrated red cell in CLCVP group was higher than that of in NCVP group(P<0 .05) .Differences between the two groups in postoperative hospital stay were with no significance in all the operation types(P>0 .05) .Conclusion The efficiency of CLCVP on blood protection during hepatectomy is influenced by the sur‐gery type ,the blood protection is found to be significant only in irregular partial hepatectomy .No relationship was found between CLCVP and postoperative hospital stay in all types of hepatectomy .

OBJECTIVETo investigate the value of susceptibility weighted imaging(SWI)in the diagnosis of hemorrhagic foci early after blast injury and its role in the outcome prediction.

METHODSTotally 30 rabbits with blast-induced cerebral blast injury were used in this study. After routine CT/MRI and SWI scanning,quantified analysis was performed in regions of interest using post-processing technology. After dissecting the brains of the experimental rabbits,the cerebral histopathological changes were observed,and the results were compared with SWI findings.

RESULTSIn these 30 rabbits,22,102,221,and 738 hemorrhagic foci were detected by CT,T1WI,T2WI,and SWI,respectively. The number of cerebral microbleeds detected by SWI was significantly larger than those revealed by conventional T1WI and T2WI(Χ(2)=10.00,P<0.01). Furthermore,the SWI imaging displayed the punctiform(n=315,42.7%),lamellar(n=218,29.5%),slinar(n=205,27.8%)hypointense foci,with clear margin. The number of hemorrhagic foci detected by SWI was positively correlated with survival(r=-0.667,P<0.05).

CONCLUSIONSSWI remarkably increases the detection rate of hemorrhagic foci(particularly microbleeds)in rabbits with cerebral blast injury. The number of cerebral microbleeds and location of foci are closely related with the outcomes and therefore may facilitate clinical managment.

Animals ; Blast Injuries ; complications ; diagnosis ; Brain ; pathology ; Brain Injuries ; diagnosis ; etiology ; Cerebral Hemorrhage ; diagnosis ; etiology ; Female ; Image Enhancement ; Magnetic Resonance Imaging ; methods ; Male ; Prognosis ; Rabbits

Objective To understand the prevalence and disease distribution of migrants within the water source area of the Middle Route Project of South-to-North Water Diversion Project and analyze the influencing factors. Methods By multi-stage stratified cluster sampling method, 307 immigrants and 676 immigrants were selected from Danjiangkou reservoir area, Shiyan city, Hubei Province. Results The prevalence of chronic diseases among in-migration was 51.8%. The prevalence rates of the top five chronic diseases were waist and leg pain, hypertension, gastritis, heart disease and diabetes. The prevalence of chronic diseases among out-migration was 65.1%. The prevalence rates of the top five chronic diseases were waist and leg pain, hypertension, gastritis, heart disease and bronchitis. The rate of chronic disease, the rate of two weeks and the rate of two weeks' visit were higher than that of emigration(all P<0.05). The results of multivariate analysis showed that the factors influencing the chronic diseases of immigrants were 60 years old (OR=59.928，P<0.001), unsatisfied family income (OR=0.377, P=0.002) and poor economic status (OR=0.517, P=0.028). The factors influencing the chronic diseases of immigrants were 60 years old (OR=26.190, P<0.001), smoking (OR=1.856,P=0.004), unsatisfactory family income (OR=0.598,P=0.029) and poor economic status (OR=0.460,P<0.001). Conclusions The prevalence of migrants within a high level is at a high level. Actually raising the income of immigrants is conducive to improving the health condition of immigrants.

OBJECTIVETo evaluate the estrogenic activity of several kinds of Chinese herbal medicines.

METHODUse zoopery and reporter gene technique to study the estrogenic activity of five Chinese herbal medicines. Zoopery: weanling female Kunming mice weight 9-12 g were administrated botanical extracts of Selaginella tamariscina, Pinus Massoniana, Corallodiscus flabellate, Dryopteris sublaeta and Leonurus heterophyllus, the positive control group with Nilestriol tablets and control group with water, respectiely. On the eighth day, the animals were sacrificed and the uteri were separated solely and weighed. Reporter gene technique: Induce the expression of reporter gene controlled by ERE and measure the activity of luciferase on cell's clear supernatant.

RESULTThe botanical extracts of S. tamariscina can increase weights of mice (P < 0.01); In the expression of reporter gene controlled by ERE, Either ERalpha or ERbeta's has estrogenic activity (P < 0.01). Follow in the zoopery we find the water part and the n-butanol part of S. tamariscina are the two active parts.

CONCLUSIONS. tamariscina and it's water part and n-butanol part have estrogenic activities, effect on ERbeta is greater than ERalpha.

Animals ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Estrogen Receptor alpha ; metabolism ; Estrogen Receptor beta ; metabolism ; Female ; Leonurus ; chemistry ; Mice ; Organ Size ; drug effects ; Phytoestrogens ; isolation & purification ; pharmacology ; Pinus ; chemistry ; Plants, Medicinal ; chemistry ; Selaginellaceae ; chemistry ; Uterus ; anatomy & histology

OBJECTIVETo establish the quality standard of PsL injections containing mainly 5F (ent-11alpha-hydroxy-15-oxo-kaur-16-en-19-oic-acid).

METHODThe identification of PsL was performed by thin-layer chromatography, and the content was determined by HPLC. The column was Hypersil C18 (4.6 mm x 250 mm, 5 microm), the mobile phase was the mixture of methane-water-acitic acid (55:45: 0.045) with a flow rate of 1.0 mL x min(-1), the detective wavelength was 254 nm, and the column temperature was maintained at 35 degrees C. The pH value and K+ content of the three batchs injection were determined with pH meter and flame photometric meter, and the contents of tannin, protein, oxalic acid salt and heavy metals were detected by deferent methods.

RESULTThe TLC method was suitable for the identification of PsL5F. The linearity for 5F was obtained over the range of 30-240 microg x mL(-1) (r = 0.999 8), the average recovery of 5F was 99.8%. The injections were of pH value range from 7.80 to 8.20, K+ contents less than 10 mmol x L(-1), and the contents of tannin, protein, oxalic acid salt and heavy metals were qualified with the Chinese pharmacopoeia, respectively.

CONCLUSIONIt's sensitive and reliable that can be used as quality control methods of PsL5F injections.

Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Diterpenes ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Injections ; Reproducibility of Results

OBJECTIVETo evaluate the effect of cytomegalovirus (CMV) infection following kidney transplantation on long-term renal function and its mechanism.

METHODSNinety-six patients undergoing kidney transplantation between March 2000 and December 2005, who completed a 3-year follow-up investigation, were divided into 3 groups according CMV-pp65 antigenemia and clinical symptoms. Group A consisted of 33 recipients with symptomatic active CMV infection, group B included 33 with asymptomatic active CMV infection and group C included 30 with inactive infection. The relation of CMV infection, transforming growth factor-beta1 (TGF-beta1) mRNA in the peripheral blood mononuclear cells (PBMCs) and serum creatinine (Scr) were analyzed, and the grafts in 6 cases with renal dysfunction were biopsied.

RESULTSThe expression of TGF-beta1 mRNA in PBMCs was significantly higher in group A than in the other two groups 6 months after the transplantation (P<0.01), while Scr levels showed no significant difference between the 3 groups (P>0.05). Three years later, Scr levels in group A were significantly increased as compared with those in the other two groups (P<0.01), and the rate of renal dysfunction in group A (10/33) was significantly higher than those in group B (3/33) and C(3/30) (P<0.05). In the 16 with renal dysfunction, the expression of TGF-beta1 mRNA in PBMCs significantly higher than that in the other 80 patients with normal renal function (P<0.01). Renal allograft biopsies demonstrated mild or severe interstitial fibrosis, tubular atrophy and mononuclear cell infiltration in the 6 patients with renal graft dysfunction, supporting the diagnosis of chronic allograft nephropathy (CAN).

CONCLUSIONSymptomatic active CMV infection in renal allograft recipients is an important factor contributing to the occurrence of CAN. Monitoring of TGF-beta1 mRNA expression in PBMCs proves useful in identifying patients at risk of CAN.

Adult ; Creatinine ; blood ; Cytomegalovirus Infections ; blood ; metabolism ; physiopathology ; Female ; Humans ; Kidney ; metabolism ; physiopathology ; virology ; Kidney Transplantation ; Leukocytes, Mononuclear ; metabolism ; Male ; RNA, Messenger ; genetics ; metabolism ; Transforming Growth Factor beta1 ; genetics ; Transplantation, Homologous

Objective To create a simple differential diagnostic rule for tuberculous meningitis and cryptococcal meningitis in adults on the basis of basic clinical and laboratory features. Methods We compared the clinical and laboratory features of 219 adults admitted to our hospital during the period of 2000-2008; these patients satisfied the diagnostic criteria for tuberculous meningitis (n=100) and cryptococcal meningitis (n=119). Multivariate logistic regression analysis was employed to find the risk factors those could independently predict the tuberculous meningitis,so that the diagnostic rule could be created. Results Six risk factors those could independently predict the tuberculous meningitis were found, including gender, mind disorder, vision and/or hearing damage, amount of cerebrospinal fluid protein, total cerebrospinal fluid white-cell count and combination with periphery tuberculosis. A diagnostic rule developed from these features enjoyed a 78.0% sensitivity, a 95.2% specificity, a 92.9% positive predictive value and a 84.4% negative predictive value. Conclusion Simple clinical and laboratory features can help in the differential diagnosis between adult tuberculous meningitis and cryptococcal meningitis, which can be applied in primary hospital with limited microbiological resources.

OBJECTIVETo investigate the molecular mechanisms of icariin (ICA) in regulating the bone formation of osteoblasts and the bone resorption of osteoclasts.

METHODSPrimary osteoblast cell cultures were obtained from newborn rat calvarial. Calcified nodules were stained by alizarin red. The mRNA levels of osterix (OSX), runt-related transcription factor 2 (Runx-2), alkaline phosphatase (ALP), Collagen1, osteoprotegerin (OPG), and receptor activator of nuclear factor-ΚB ligand (RANKL) were analyzed by quantitative real-time RT-PCR, the protein levels of OPG, RANKL, and Collagen1 were examined by Western blotting, and the intracellular Ca(2+) concentration of osteoblasts was measured on a flow cytometer using the Cellquest program.

RESULTSCompared with control group, ICA markedly promoted bone formation by significant up-regulating the gene expressions of OSX, Runx-2,ALP, and Collagen1, the protein expression of Collagen1(all P<0.01), and the Ca(2+) concentration. Furthermore, ICA remarkably inhibited bone resorption by significant up-regulating the mRNA and protein expressions of OPG as well as the OPG/RANKL ratio.

CONCLUSIONSICA could promote bone formation of osteoblasts through inducting the gene expressions of OSX,Runx-2, ALP and Collagen1, and the protein expressions of Collagen1, and by increasing the Ca (2+) concentration. Moreover, ICA could inhibit bone resorption of osteoclasts through regulating OPG/RANKL signal pathway.

Alkaline Phosphatase ; metabolism ; Animals ; Bone Resorption ; Cells, Cultured ; Collagen Type I ; metabolism ; Core Binding Factor Alpha 1 Subunit ; metabolism ; Flavonoids ; pharmacology ; Gene Expression ; Osteoblasts ; drug effects ; Osteoclasts ; drug effects ; Osteogenesis ; drug effects ; Osteoprotegerin ; metabolism ; RANK Ligand ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptor Activator of Nuclear Factor-kappa B ; metabolism ; Transcription Factors ; metabolism