Objective To study the effect of human Slitrk1 gene on proliferation and differentiation of PC12 cells. Methods The CDS sequence of Slitrk1 was amplified by PCR, and then cloned into pcDNA4 vector. The recombinant plasmid pcDNA4/Slitrk1 were transfected into rat PC12 cells by lipofectamine. The stable expression cell clones were screened by RT-PCR. MTT method was used to detect the proliferation rate of PC12 cells. The morphologic changes in PC12 cells were observed microscopically. Results The stable cell lines expressing pcDNA4 (pcDNA4/PC12) and pcDNA4/Slitrk1 (ST1/PC12) were established. Compared to pcDNA4/PC12 cells, the growth rate of ST1/PC12 cells was decreased. In addition, pcDNA4/PC12 cells tend to grow as well as the normal PC12 cells. However, most of the ST1/PC12 cells adhered to the plate with one or two neurites. Conclusion Over-expression of human Slitrk1 gene inhibited the proliferation of PC12 cells and promoted the outgrowth of neurites. It is suggested that human Slitrk1 gene may be involved in differentiation of PC12 cells.

Objective To define laboratory technology for expression and purification of nucleocapsid proteins of human coronaviruses SARS-CoV,HCoV-229E,HCoV-OC43,HCoV-NL63 and HCoV-HKU1,animal coronaviruses bovine coronavirus,murine hepatitis virus,feline infectious peritonitis virus,and infectious bronchitis virus.MethodsThe pET-30a-based recombinant plasmids containing full length nucleocapsid(N) protein of coronaviruses were transformed into E.coil BL21(DE3) competent cells and were induced to express N proteins by IPTG.The expression products were purified by ion exchange chromatography and Ni2+ affinity chromatography,and were verified by SDS-PAGE and Western blot.Results Nine coronavirus nucleocapsid proteins with correct Mr were solubly expressed and highly purified with purity over 90%.Conclusion Successfully expressed nine recombinant nucleocapsid proteins with high purity in E.Coli,which provides materials to study the function of these coronavirus N proteins.

Objective:To investigate the effect of anti-CD134 mAb or CTLA4Ig on ConA induced splenic cell proliferation,Th cytokine secretion and production of anti-dsDNA antibody from splenic lymphtocyte in vitro in lupus-prone BXSB mice. Methods:Eighteen male lupus-prone BXSB mice model and 6 syngeneic normal C57BL/6 male mice were used in the experiment. The model mice were divided into three groups:un-treated group,Lupus recipe(LR) treated group and prednisone(pred. ) treated group. The mice's splenic cell suspension from above groups was culture stimulated by ConA respectively. The splenic cells from un-treated model mice were further divided into Anti-GD134L mAb,CTLA4Ig or Anti-CD134L mAb + CTLA4Ig treated subgroups. The ConA induced splenic cell proliferation was measured by MTT colorimetric assay. The levels of IFN-?, IL-6 and anti-dsDNA antibody in cell supernatant were measured by ELISA. Results; (1 )The splenic cell proliferative reaction and contents of IFN-?,IL-6 and anti-dsDNA antibody in cell supernatant of either spontaneous or ConA induced culture in the un-treated model group were obviously higher than that of the normal control or other groups. (2) The splenic cell proliferative reaction and production of IFN-?,IL-6 and anti-dsDNA antibody in the CD134L/CTLA4Ig treated group,LR treated goup or pred. treated group was not different from the normal control significantly. (3)To compared with CD134L treated group or CTLA4Ig treated gruop,the CD134L/CTLA4Ig and prednisone reduced significantly the splenic cell proliferative reaction and production of IFN-?,IL-6 and anti-dsDNA antibody in cell supernatant of either spontaneous or ConA induced culture,while no difference was found between CD134L treated group and CTLA4Ig treated proup. Conclusion:The lupus-prone BXSB mice might present abnormal lymphocyte proliferation,spontaneously express cytokines and secrete high level of autoantibody during the SLE development. LR and corticosteroids could obviously inhibit the abnormal lymphocyte proliferation;reduce the Th cytokine formation and antoantibody production Blockade of CD134-CD134L or B7-CD28 costimulatory pathway by Anti-CD134L rnAb or CT-LA4Ig could inhibit the activation of T cells and B cells like LR and corticosteroids. Furthermore, by blockade of both CD134-CD134L and CD28-B7 pathways,the frequency of alloreactive T cell was markedly reduced and was maintained at low levels so as to treat SLE effectively.

AIM: To investigate the effects of lupus recipe on immune system and lymphocyte subsets proliferation in splenic cells in BXSB mice. METHODS: Eighteen male BXSB mice model was used in the experiment. The model mice were divided into three groups: un-treated model group, lupus recipe (LR) treated group, and prednisone treated group. All model mice were killed in 10 weeks. The control group consisted of 6 syngeneic normal C57BL/6 male mice. The levels of total IgG and anti-dsDNA antibody in serum were detected by ELISA. The percentages of lymphocyte subsets (CD3+, CD4+, CD8+ T lymphocytes and CD19+, CD23+ B lymphocytes) were detected by using flow cytometry analysis. RESULTS: (1) The serum levels of total IgG and anti-dsDNA antibody in un-treated model group were higher than that in other groups. There was no differences among LR treated group, prednisone treated group and control group. (2) The percentages of CD3+, CD4+, CD8+ T lymphocytes and CD19+, CD23+ B lymphocytes in model group were obviously higher than that in normal control. (3) Compared to un-treated model group, the percentages of CD4+, CD8+ T lymphocytes and CD19+, CD23+ B lymphocytes in LR or prednisone treated group were significantly reduced, which closely reached the levels in normal group. CONCLUSIONS: The immune functions of T and B lymphocytes in BXSB mice are up-regulated. LR inhibits the activation of T and B lymphocytes, reduces the serum levels of IgG and auto-antibody production.

Objective To effectually record cardiopulmonary resuscitation (CPR) procedure, analyze and compare the CPR performance of all medical and nursing staffs to find out the existed information for the improvement of the quality of CPR and its survival rate. Methods The medical data were collected according to the Utstein Criteria and CPR event was automatically recorded by a digital video-recording system, by which hands-off times within 10 minutes of CPR and times for installation of chest compression machine, establishment of endotracheal intubation and establishment of the venous channel were analyzed. Multiple regression analysis was conducted to analyze the factors affecting CPR effect. Results ① During the period from December 2009 to December 2015, a total of 376 patients with cardiac arrest (CA) was registered, including 248 males and 128 females, with a median age of 68 (53, 78) years. Estimated median time interval from CA to CPR initiation was 5.0 (0.1, 20.0) minutes and there were 189 cases less than or equal to 5 minutes. The acute myocardial infarction (AMI) with 145 cases (38.6%) was the main etiology of CA. Initial shockable rhythm was found in 16 patients (7.0%) out of 230 out-of-hospital cardiac arrest (OHCA) patients, of whom 13 underwent ventricular fibrillation (5.7%), 3 underwent ventricular tachycardia (1.3%). Initial shockable rhythm was found in 47 patients (32.2%) out of 146 in-hospital cardiac arrest (IHCA) patients, of whom 40 underwent ventricular fibrillation (27.4%), and 7 underwent ventricular tachycardia (4.8%). CPR by a mechanical device (Thumper) was performed in 219 patients (58.2%). In 376 patients, 186 patients had return of spontaneous circulation (ROSC, 49.5%), a successful CPR (ROSC ≥ 24 hours) was found in 110 patients (29.3%), 99 patients was hospitalized alive (26.3%) and 40 patients were discharged alive (10.6%). In 146 cases of IHCA, 89 patients had ROSC (61.0%), a successful CPR was found in 63 patients (43.2%), 56 patients were hospitalized alive (38.4%), and 29 patients were discharged alive (19.9%). In 230 patients of OHCA, 89 patients had ROSC (38.7%), 65 patients received pre-hospital CPR (28.3%), a successful CPR was found in 47 patients (20.4%), 43 patients were hospitalized alive (18.7%), and 11 patients were discharged alive (4.8%). There were 37 patients had a successful CPR (69.8%), and 25 patients were discharged alive (47.2%) in 53 patients with ventricular fibrillation. ② There were 77 patients with valid video information for analysis of CPR performance, with 48 patients of OHCA, and 29 patients of IHCA. Delay median time from the patients presence in the resuscitation room to be placed in rescue bed was 22 (0, 33) seconds. Hands-off median times during 10 minutes of CPR was 41 (18, 90) seconds. Thumper installment median times was 43 (31, 69) seconds. Median time for endotracheal intubation was 59 (35, 109) seconds. Median time of venous catheter placement was 112 (70, 165) seconds. It was shown by multivariate regression analysis that there was a significant correlation between estimated time interval from CA to CPR performed, hands-off time and success rate of CPR (t1 = -3.452, t2 = -2.729), rate of discharge alive (t1 = -2.328, t2 = -2.736, all P < 0.05). In 48 OHCA patients, success rate of CPR was significantly correlated with estimated time interval from collapse to CPR performed (t = -2.409, P = 0.021). In 29 IHCA patients, success rate of CPR and rate of discharge alive was significantly correlated with hands-off times (t1 = -3.412, t2 = -2.536, both P < 0.05). Conclusions Survival to hospital discharge following CA is significantly correlated with the time interval from collapse to CPR performed and hands-off times in CPR. Installment and usage of Thumper should be postponed in order to reduce hands-off times during CPR in IHCA.

A large number of documents will be generated in the process of new drug research,including lots of references.These data accumulate into institutional repository.Up to now,the management of document knowledge base in the process of new drug research has not yet been published.The paper uses the reference management software,combined with the document knowledge management system,to analyze and research the management model and method for establishing institutional repository.

Objective:To observe the hemodynamic change and inhibitory effect on stress response of oxycodone during induction, maintenance and resuscitation period of general anesthesia in patients undergoing gastric cancer surgery.Methods:60 patients with general anesthesia undergoing elective gastric cancer from March 2018-March 2019 in No.904 Hospital were selected. According to the random number table method, they were randomly divided into two groups ( n=30): Oxycodone group (group O) and Fentanyl group (group F). The anesthesia was induced by intravenous oxycodone 0.3 mg/kg (group O) or fentanyl 3 μg/kg (group F), propofol 2 mg/kg and cisatracurium 0.25 mg/kg. Oxycodone (group O) or fentanyl (group F) with sevoflurane were used for maintenance of general anesthesia. The mean arterial pressure (MAP) and heart rate (HR) were recorded before anesthesia induction (T 0), immediately intubation (T 1), 1 min (T 2) and 5 min (T 3) after intubation, immediately skin incision (T 4), surgery for 1 h (T 5), the end of operation (T 6) and 1 min (T 7) and 5 min (T 8) after extubation. The concentration of glucose (Glu), cortisol (Cor), epinephrine (E) and norepinephrine (NE) were detected in time T 0,T 2, T 4,T 5 and T 7. The time of extubation and the visual analogue scale (VAS) scores after extubation and 30 min after entering postanesthesia care unit (PACU) were recorded. The incidence of cough during induction, dysphoria during extubation and the adverse effects such as nausea, vomit and respiratory depression during PACU were also observed. Results:There were no significant difference between two groups in MAP and HR at the time point from T 0 to T 8 ( P>0.05); there were no significant difference about concentrations of Glu, Cor, E and NE at the time point of T 0,T 2,T 4,T 5 and T 7 ( P>0.05). Extubation time and VAS scores of 30 min after entering PACU in group O were significantly lower than those of group F ( P<0.05). Meanwhile, the incidences of cough during induction, dysphoria during extubation and respiratory depression in group O were significantly lower than in group F ( P<0.05). There was no significant difference in the incidence of nausea and vomiting ( P>0.05). Conclusions:Oxycodone can be safely and effectively used for induction and maintenance of general anesthesia in gastric cancer surgery, inhibit intraoperative stress response, and improve the quality of recovery period.

OBJECTIVE:To study the purification technology of total flavonoids from Nelumbinis receptaculum by macropo-rous resin. METHODS:Using adsorption rate and desorption rate of total flavonoids from Nelumbinis receptaculum as index,the type of macroporous resin was selected by static adsorption-desorption tests;With adsorption rate of total flavonoids as index,sin-gle factor test was used to investigate the effects of the concentration of total flavonoids,adsorption time,adsorption speed, drug-loading amount,water amount,volume fraction and amount of eluant and other factors on the purification technology. The op-timal technology was validated. RESULTS:Among 10 kinds of resin,HPD-400 macroporous resin was found to have the best ad-sorption and desorption effects. The optimal purification conditions was as follows as the concentration of total flavonoids 7.00 mg/ml, adsorption time of 3 h,flow rate for sampling of 3 column volume (BV)/h,drug-loading amount of 8 BV,water amount of 6 BV,50% ethanol elution amount of 4 BV. In validation test,mass fraction of total flavonoids from purified Nelumbinis receptacu-lum were 63.88%,62.50% and 63.44%(RSD=1.11%,n=3). CONCLUSIONS:HPD-400 macroporous resin could purify total flavonoids from purified Nelumbinis receptaculum,and established purification technology is stable and practical.

Objective To investigate the therapeutic effects of recombinant human erythropoietin(rhEPO)on brain mitochondrial energy metabolism and mitachondrial respiratory functionin after brain injury in rats.Method A total of 63 Sprague-Dawley rats were divided randomly into three groups:the rhEPO treated group(n =28),the control group(n=28),the shanl group(n=7).The models of contusion of brain caused by freefalling were set up in rhEPO treated group(n=28).The recombinant human erythropoietin was intraperitoneally injected in dose of 10 U/g immediately after brain injury and it was repeated every 10 hours in rhEPO group treated.The same models of contused brain were made without rhEPO treatment as control group(n=28).In control group,the same volume of normal saline was used in replacemem of rhEPO.Aburr hole was made on the skull of the sham group(n=7),but the brain tissue was not wounded.The mitochondria were isolated at 6 h,12 h,24h,48 h after trealment,respectively.The activity of ATPase and SOD,the content MDA and brain mitochondrial respiratory function were measured by biochemical technique.The data were analyzed with the F-test and t-test.Results The activity of ATPase(P＜0.05),SOD(P＜0.01)and brain mitochondrial respiratory function(P ＜0.05)were increased.and the levd of MDA in brain mitochondria was reduced markedly in rats treated with rhEPO.Conclusions Treatment with rhEPO can alleviates the secondary brain injury by affecting mitochondrial function.

Objective To compare the diagnostic significance of pleural SLPI,IFN-γ and ADA for differenti-ating TPE from pleural effusions with the other etiologies. Methods Pleural effusion samples were obtained from 93 patients who were divided into the following groups: tuberculous pleural effusion,malignant pleural effusion, bacterial pleural effusion and transudative pleural effusion. The pleural effusion and/or serum levels of SLPI , IFN-γand ADA were determined. Results 1.The concentrations of SLPI, IFN-γand ADA in tuberculous pleural effusion was higher than that in malignant group, bacterial group and transudative group. 2. The diagnostic value of SLPI, IFN-γor ADA for the diagnosis of tuberculous PE is high respectively. The combinations of SLPI, IFN-γand/or ADA gained the more valuable diagnostic performance. Conclusion Pleural SLPI, IFN-γand ADA may be helpful for the differential diagnosis of tuberculous pleural effusion and the other pleural effusion. The combinations of SLPI or/and IFN-γor/and ADA further increased diagnostic value.