Objective To evaluate the effect of isoflurane anesthesia on Wnt3a expression in the hippocampus of developing rats.Methods Twenty-four 7-day-old male Sprague-Dawley rats,weighing 125-155 g,were randomly divided into 3 groups (n=8 each) using a random number table:control group (group C),4 h inhalation of 2％ isoflurane group (group I4),and 6 h inhalation of 2％ isoflurane group (group I6).The rats were sacrificed at 6 h after the end of treatment in each group,and the hippocampi were removed for determination of Wnt3a mRNA expression (by real-time reverse transcriptase polymerase chain reaction) and Wnt3a expression (by Western blot).Results Compared with group C,the expression of Wnt3a and Wnt3a mRNA in hippocampi was significantly up-regulated in I4 and I6 groups (P＜0.05).Compared with group I4,the expression of Wnt3a in hippocampi was significantly up-regulated (P＜0.05),and no significant change was found in the expression of Wnt3a mRNA in hippocampi in group I6 (P＞0.05).Conclusion The mechanism of isoflurane-induced neurotoxicity is probably related to upregulation of Wnt3a expression in the hippocampal tissues of developing rats.

OBJECTIVE To explore the effect of E-cadherin on the proliferation ability of Hep-2 by method of RNA interference technology to silence the expression of E-cadherin. METHODS The specific siRNA sequences and non-silencing siRNA were designed and synthesized. Hep-2 cells were transfected and then the down expression of E-cadherin gene in vitro cultured Hep-2 cells were got. The silencing effect of E-cadherin gene was explored by Fluorescence Quantitative Polymerase Chain Reaction and the proliferation of the transfected Hep-2 cells were detected in vitro by MTT assay. RESULTS 1.When transfected with the ratio of recombinant plasmid and the quality of liposome volume at 1:1, the transfection efficiency at the siRNA-3 group was the highest and can be up to 65%. 2.The results of Fluorescence Quantitative Polymerase Chain Reaction: recombinant plasmid pRNAT-U6.1/Neo-siRNA1, pRNAT-U6.1/Neo-siRNA2 and pRNAT-U6.1/Neo-siRNA3 can down regulate the expression of E-cadherin mRAN. Set blank control group as a baseline (set to 1), the changes of expression of E-cadherin relative to β-actin in siRNA-1group was 0.00092, siRNA-2 group was 0.00143, siRNA-3 group was 0.00045 and the negative control group was 3.44898. The difference was statistically significant (P<0.05). 3. MTT: The growth rate of Hep-2 cells treated by specific siRNA was faster than that of the control group, and the difference was statistically significant (P<0.05). CONCLUSION Effectively inhibition the expression of E-cadherin's mRAN can enhance the proliferation of Hep-2 cells.

Objective To express and purify recombinant Tp0136 epitope fragment, and study the immunity activity. Methods The Tp0136 selective fragment(Tp0136B) gene was devised by the surface property analysis, solvent-accessible suface calculateions, secondary structure function region analysis, and was inserted between the sites of Nde Ⅰ and Not Ⅰ in pET22b ( + ) . The recombinant plasmid was expressed in E. coli BI21. After nickel ion metal affinity chromatography, the antigenic and immune reactivity of rTp0136B was confirmed. Then indirect ELISA with the rTp0136B as coating antigen was performed to detect the anti-Tp0136 antibody in sera from 100 normal human controls and 131 primary syphilis patients. Results The rTp0136B was soluble expressed with a molecular weight of about 28 000 and was obtained with a purity of ＞98% by chromatography. Western blot proved that the rTp0136B could specifically react with anti-Tp0136 polyclonal antibody. Specific humoral response was elicited by the recombinant protein in Japan negative. The positive detection rate in sera from primary syphilis patients was 85.5%. Conclusion This result suggested that the recombinant Tp0136 epitope fragments have a satisfactory immunocompetence,which may have applications in the serodiagnosis of primary syphilis.

Adolescent ; Female ; Humans ; Patellar Dislocation ; physiopathology ; surgery ; Patellar Ligament ; injuries ; physiopathology ; surgery ; Range of Motion, Articular ; Reconstructive Surgical Procedures

OBJECTIVETo explore material bases and neurobiological mechanisms of "Shen storing will" by observing learning and memory capacities and N-methyl-D-aspartic acid (NMDA) receptor expressions in Shen deficiency constitution (SDC) rats.

METHODSTotally 40 SD rats were randomly divided into the model group, the Zuogui Pill (ZP) group, the Yougui Pill (YP) group, the blank control group (consisting of normal pregnant rats), 10 in each group. SDC young rat model (inherent deficiency and postnatal malnutrition) was prepared by the classic way of "cat scaring rat". Medication started when they were scared by cat. Rats in the ZP group and the YP group were administered by gastrogavage with ZP suspension 0.1875 g/mL and YP suspension 0.0938 g/mL respectively. Equal volume of normal saline was administered to rats in the blank control group and the model group by gastrogavage. All medication was given once per day, 5 days in a week for 2 consecutive months. Learning and memory capacities were detected by Morris water maze test. Expressions of NMDA receptor subunits NR2A and NR2B in hippocamus were detected by immunohistochemical method.

RESULTSCompared with the blank control group, the latency period, total distance in Morris water maze test were longer in the model group (P < 0.05). All the aforesaid indices all decreased in the ZP group and the YP group, with statistical difference when compared with the model group (P < 0.05). The protein expressions of NR2A and NR2B in hippocamus were lower in the model group than in the blank control group (P < 0.05). But when compared with the model group, they were obviously higher in the ZP group and the YP group (P < 0.05).

CONCLUSIONSSDC rats had degenerated learning and memory capacities and lowered NMDA receptor expressions. ZP and YP could up-regulate learning and memory capacities and NMDA receptor expressions, thereby improving deterioration of brain functions in SDC rats.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Learning ; drug effects ; Memory ; drug effects ; Pregnancy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism ; Up-Regulation

Objective To investigate the effect of acupuncture combined with minimally invasive hematoma aspiration on a rabbit model of brain injury in the acute stage of intracerebral hemorrhage (ICH). Methods A total of 65 healthy New Zealand white rabbits were randomly divided into a sham operation (SO) group, an ICH group,a hematoma aspiration (HA) group, and an acupuncture and hematoma aspiration (AHA) group. Models of ICH were established in the latter three groups. The HA group was treated with minimally invasive hematoma aspiration and the AHA group was treated with both acupuncture and minimally invasive hematoma aspiration. At 6 hours and 1, 3 and 7 days after the ICH models were established, brain water content (BWC) was measured, and the expression of matrix metalloproteinase-9 ( MMP-9 ) in the rabbits' brains was detected by immunohistochemistry. MMP-9activity was detected by gelatin zymography. Results The BWCs of the ICH group, HA group and AHA group rabbits were significantly higher than those of the SO group. The BWCs of the HA group and AHA group animals descended significantly more than those in the ICH group as time went on, especially on the 3rd day in the AHA group. Immunohistochemistry and gelatin zymography showed that the expression and activity of MMP-9 in these test groups decreased with time, especially on the 3rd day in the AHA group. Conclusions Acupuncture combined with hematoma aspiration can reduce injury in the acute stage of ICH, and inhibition of the expression of MMP-9 may be the mechanism.

Iridoid glycosides were the main active ingredient of Rehmannia glutinosa, of which catalpol has the highest content. This work will provide theoretical basis for metabolic study and cultivation of iridoids on the basis of the dynamic accumulation of catalpol and total iridoids in the growth of R. glutinosa. The samples of rehmannia 85-5 were gathered in the same filed from July to October. The contents of catalpol and total iridoid glycosides were measured by HPLC and specteophotometric, respectively. The results showed that youngest leaves had the higher content of catalpol and total iridoid glyosides than that of the other two leaf ages in the same growth stage from July to September, while their content of catalpol and total iridoid glycosides were all decreased as the growth of leaves of R. glutinosa. The content of catalpol didn't differ significantly from July to September, whereas it has significantly increased in October in the three leaf stage. In the same stage, the wider the root diameter is, the higher content of the effective components are. In August and September, the total iridoid glycosides have the fastest accumulation. The content of catalpol was increased as the accumulation of total iridoid glycosides.
Iridoid Glucosides ; metabolism ; Iridoids ; metabolism ; Plant Roots ; metabolism ; Rehmannia ; growth & development ; metabolism ; Seasons ; Water ; metabolism

Objective To evaluate the clinical efficacy of a stroke unit combined with community health services for treating stroke survivors. Methods A total of 120 stroke patients were randomly divided into a " stroke unit combined with community medicine" group ( combined group) , a stroke unit group and a general treatment group. Patients in the former 2 groups were treated in a hospital stroke unit during their hospitali-zation. The general treatment group was given conventional medical treatment. After discharge, the combined group continued to receive regular rehabilitation therapy and guidance in the form of community medical services, while the stroke unit group received follow-up only. Assessment was by means of Fugl-Meyer scores, the Barthel index and self-rating on a depression scale ( SDS). The patients were assessed at admission, on discharge and 3 months after discharge. Results There were no significant differences in average limb motor function, ability in the activities of daily living ( ADL) or depressive mood among the 3 groups on admission, but at discharge, limb motor function and ADL ability in the combined group and stroke unit groups were significantly superior to those in the general therapy group. Limb motor function and ADL ability in the combined and stroke unit groups had improved further 3 months after discharge, with more significant improvements in the combined group. No significant change in depression was observed in any group at discharge, but average depression scores in the combined and stroke unit groups improved significantly in the 3 months after discharge, and there was a statistically significant difference between the combined group and the general group. Conclusion Supplementing the work of a stroke unit with community health services significantly improves stroke patients' recovery of limb motor function and ADL ability.

Objective To detect the expression of delta-like ligand 4 (DLL4) and its relation with the angiogenesis in pancreatic cancer.Methods SP immunohistochemistry method was used to detect the expression of DLL4 in pancreatic cancer and para-cancerous tissue.The microvessel density (MVD) was calculated with CD34 staining for microvessel endothelium and the correlation between expression of DLL4,MVD and clinicopathological parameters of pancreatic carcinoma was investigated.Results The expression of DLL4 in pancreatic cancer was significantly higher than that in para-cancerous normal pancreatic tissue (68.3％ vs 20％,P ＜0.01 ).The over-expression of DLL4 in pancreatic cancer was positively related to the differentiation degree,TNM staging,metastasis and invasion (P ＜0.05),but not related to the location,tumor size and histological types.The MVD in pancreatic cancer tissues was significantly higher than that in para-cancerous normal tissues (34.9 ± 13.2 vs 18.9 ±2.2,P ＜0.01).MVD was correlated with the differentiation degree,TNM staging,metastasis and invasion (P ＜0.05 ),but not with the tumor size and location,histological types.MVD in pancreatic cancer expressing DLL4 was significantly higher than that in pancreatic cancer not expressing DLL4 ( 38.8± 10.7vs 29.0 ± 15.2,P =0.038 ),and the expression of DLL4 and MVD were positively correlated ( r =0.669,P ＜ 0.05 ).Conclusions DLL4 expression may play an important role in pancreatic carcinogenesis by promoting angiogenesis and may be associated with tumor invasion and metastasis.

Objective To analyze the distribution of multiple risk factors for hospitalized patients with acute coronary syndromes (ACS) and explore the status and determinants of drug usage recommended by the guideline.Methods This was a multi-center cross-sectional study in 34 hospitals from 22 provinces in China.About ninety ACS patients were consecutively enrolled from each hospital since April 15,2012 according to a standard protocol.Totally,3 253 patients with complete data were analyzed in this study.Results (1) The average age of male patients with ACS was lower than that of female patients (60.4 years vs 66.2 years,P ＜0.01).Nearly 60％ of ACS patients were under the age of 65 years.Early onset of ACS accounted for one-third of male (＜ 55 years of age) and two-fifthes of female patients (＜ 65 years old).(2) Among the four ACS major risk factors (hypertension,hyperlipidemia,smoking and diabetes),hypertension was with the highest prevalence (68.4％).More than 90％ of ACS patients had at least one risk factor and about two-thirds of them had at least two.(3) As for the application of evidence-based drugs,the top one was aspirin with 95.3％ of ACS patients reseiving it.The second was statins (90.1％).Angiotensin converting enzyme inhibitors (ACEI) or angiotension Ⅱ receptor blocker(ARB) was the lowest (53.6％).Multivariable analysis indicated that,in contrast to that in ACS patients without percutaneous coronary intervention(PCI),the drug usage rates were increased by more than 30％ for ACEI or ARB and β receptor blockers,by more than 50％ for statins,and by 4-7 times for antiplatelet agents among ACS patients with PCI.Conclusions In China,more than 90％ of hospitalized patients with ACS carried at least one major risk factor.There is still room for improving in the application of drugs recommended by the guidelines,especially for ACS patients without PCI.