Objective::To explore the effects of the biomimetic network membrane prepared by chitosan/gelatin/pectin on the proliferation and mineralization of mesenchymal stem cells (MSCs) , and to evaluate its feasibility of constructing tissue engineering bone.Methods:Chitosan, gelatin and pectin were made into a new biomimetic network membrane in a certain ratio by biomimetics.The experiment was divided into control group (MSCs+conventional medium) , material group (MSCs+network membrane+conventional medium) and material+OS group (MSCs+network membrane+OS medium) .The cell morphology was observed by inverted phase contrast microscope;the growth and secretion of extracellular matrix of the MSCs were observed under scanning electron microscope (SEM) .The proliferation of cells was determined by MTT assay (The MSCs were divided into negative control group and material group, and they were cultivated with blank medium and medium including materials) .The expression of calcium in MSCs was detected by Alizarin Red staining.Real-time polymerase chain reaction (RT-PCR) was used to determine the expression levels of osteocalcin (OC) mRNA and osteopontin (OPN) mRNA in the MSCs.Results:The network membrane was semitransparent thin film.The MSCs were short shuttle and clustered under inverted phase contrast microscope.After cultured for 7d, the MSCs were shuttle;after cultured for 14d, the number of MSCs was increased, with pseudo feet on the membrane;after cultured for21d, the MSCs clustered with a lot of neo-formed extracellular matrix.The MTT results showed that there was no significant difference in the proliferation level of MSCs between material group and negative control group (P＞0.05) .The Alizarin Red staining results showed that the MSCs in the network membrane were dyed orange red.The RT-PCR results showed that the expression levels of OC mRNA in the MSCs in material group and material+OS group were lower on the 7th and 14th days, but on the 21th day, the expression levels were significantly increased and reached the peak;the expression level of OC mRNA in the MSCs in material group was significantly increased on the 7th day, and the expression level reached the peak on the 14th day, then fell slightly on the 21th day;compared with control group, the expression levels of OC mRNA and OPN mRNA in the cells in material group and material+OS group at different time points were significantly increased (P＜0.01) , but there were no significant differences between material group and material+OS group (P＞0.05) .Conclusion:Chitosan/gelatin/pectin biomimetic network membrane has good biocompatibility, and MSCs can grow and proliferate well on the membrane.The membrane can induce the MSCs to express mineralization-related genes and proteins without inducers.

Objective: To investigate the effect of thoracic paravertebral nerve block combined with general anesthesia in the perioperative period of elderly patients with non-small cell lung cancer (NSCLC). Methods: 50 elderly patients with NSCLC in our hospital from June 2015 to June 2018 were randomly divided into control group (n=25) and observation group (n=25). The control group was given general anesthesia, while the observation group was given thoracic paravertebral nerve block combined with general anesthesia. Postoperative recovery (recovery time, extubation time, ambulation time, feeding time) and hospitalization time, visual analogue scale (VAS) score at 2, 12, 24 and 48 hours after operation, serum tumor markers [cyfra21-1], carbohydrate antigen 125 (CA125), CA199, carcinoembryonic antigen (CEA) were counted and compared. The heart rate (HR), mean arterial pressure (MAP) [preoperative (T₁), 15 minutes (T₂), 5 minutes (T₃) after anesthesia], and the incidence of adverse reactions were recorded. Results: ⑴ Postoperative recovery and hospitalization time: hospitalization time, recovery room stay time, extubation time, out-of-bed activity time and eating time of the observation group were shorter than those of the control group (P<0.05); ⑵ Pain degree: VAS score of the observation group was lower than that of the control group at 2, 12 and 24 hours after operation (P<0.05), and 48 hours after operation VAS score of the observation group was lower than that of the control group (P<0.05). There was no significant difference between the two groups (P>0.05); ⑶ Serum tumor markers: there was no significant difference in serum cyfra21-1, CA125, CA199 and CEA levels between the two groups before operation (P>0.05). After operation, serum cyfra21-1, CA125, CA199 and CEA levels of the two groups were decreased, and the indexes in observation group were lower than that of the control group (P<0.05); ⑷ Hemodynamics: there was no significant difference in HR and MAP between the two groups at T₁ (P>0.05). HR and MAP in the two groups at T₂ were lower than those at T₁, but the levels of each index in the observation group were higher than those in the control group (P<0.05). HR and MAP in the observation group at T₃ stage returned to the level at T₁, and there was significant difference between the two groups (P<0.05); ⑸ Adverse reactions: the incidence of adverse reactions in the observation group (8.0%) was lower than that in the control group (32.0%, P<0.05). Conclusions The application of thoracic paravertebral nerve block combined with general anesthesia in total thoracoscopic lobectomy is safe and can inhibit the large fluctuation of hemodynamics in elderly patients with non-small cell lung cancer, shorten hospitalization time and post-operative rehabilitation time, alleviate post-operative pain, reduce the content of serum tumor markers, and reduce the risk of adverse reactions.

Objective To study the pharmacokinetics and detection window of clozapine and its metabolites in human blood, so as to provide experimental basis for forensic cases of identification of clozapine poisoning. Methods 29 Taiyuan Han people's elbow venous blood was collected after given oral administration of 12.5mg clozapine at different time point, in which clozapine and its metabolites were extracted with solid phase extraction (SPE) and determined by HPLC-MS-MS. The qualitative analysis was based on retention time and MRM ions. The quantitative analysis was based on an internal standard method and calibration curve. Using the 3p97 pharmacokinetic software, pharmacokinetic equation of clozapine in the blood were imitated from the C-T data, and pharmacokinetic parameters were calculated. Results The pharmacokinetics of clozapine met a two compartment open model with a first kinetics absorption. The Tmax of clozapine(CLP), demethylclozapine(DMCLP), N-oxidation-clozapine(NO-CLP) respectively were 2.96±1.32h, 8.65±3.00h, 9.31±26.38h; The Cmax of CLP, DMCLP, NO-CLP respectively were 34.68±9.32ng/mL, 11.16±4.15ng/mL, 9.62±13.88ng/mL;The t1/2 of CLP, DMCLP, NO-CLP respectively were 17.02±23.63h, 27.06±12.58h, 41.27±29.75h; The detection window of CLP, DMCLP, NO-CLP respectively were 81.72±26.19h, 93.21±29.40h and 19.93±14.62h. Conclusion The pharmacokinetics of clozapine in blood of Han people is consistent with two compartment open model with a first kinetics absorption. The pharmacokinetics model and parameters of clozapine can provide expirimental basis for forensic identification of clozapine poisoning cases.

Objective To study the toxicokinetics of Cymermethrin and its metabolites in dog bile and provide evidence for forensic cases of identification of Cymermethrin poisoning. Methods 1/4LD50 doses of Cymermethrin were given to 6 male dogs by oral perfusion after the gallbladder fistula surgery on them,and their bile were collected at different time, in which Cymermethrin and its metabolites were extracted by Liquid-liquid extraction with dichloromethane and detected by HPLC-MS-MS. The qualitative analysis was based on retention time and MRM ions. The quantitative analysis was based on an internal standard method and calibration curve. Toxicokinetics equations of Cymermethrin and its metabolites in the bile were established from the c-t curves which were fitted by the WinNonlin toxicokinetics software meanwhie toxicokinetics parameters were obtained. Results The toxicokinetics of Cymermethrin met first-order dynamic equation. The Tmax of Cymermethrin(CYM), 3-phenoxybenzoic acid (3-PBA), 3-(2,2-Dichloroethenyl)-2,2-dimethyl-cyclopropanecarboxylate (DCVA) respectively were 1.52±0.30,1.29±0.04,0.93±0.41 h ; The Cmax of CYM, 3-PBA, DCVA respectively were 0.38±0.03,7.9±1.32,30.9±16.24 μg/mL ; The T1/2 of CYM, 3-PBA, DCVA respectively were3.93±0.71,1.36±0.11,4.49±2.81 h; Conclusion The toxicokinetics of Cymermethrin in dog bile met first-order dynamic equation ; The toxicokinetics model and parameters of Cymermethrin can provide evidence for forensic identification of Cymermethrin poisoning cases.

Objective Study on the stability of carbofuran and its metabolite carbofuran phenol in blood preserved at different conditions,in order to provide a scientific evidence for forensic identification of carbofuran poisoning death. Methods The dogs were given intragastric administration with 4LD50(13.5mg/kg) of carbofuran, the blood were collected and divided into five equally groups preserved at 20℃(NC2.5mg/mL), 20℃(1%NaF), 20℃, 4℃ and -20℃, respectively. The concentrations of carbofuran and carbofuran phenol in above samples were detected by GC-MS/MS with MRM at 0d、5d、7d、15d、40d、83d and 150d. Results The concentration of carbofuran in preserved blood were found to be significant decrease at 7d(P < 0.05), then a steady decline. In each condition, the concentration of carbofuran phenol in preserved blood showed an increasing trend firstly, then a declined tendency. The concentration of carbofuran and carbofuran phenol descending fast in blood at 20 ℃ (NC) and 20 ℃ (1%NaF).Conclusion Carbofuran and carbofuran phenol in preserved specimens are found to be decomposed. The decomposition is quick at 20℃ and slow at -20℃. Citrate sodium and sodium fluoride are not suit for anticoagulation and antiputrefactiva. Biological specimens used for forensic identification of the carbofuran poisoning should be stored at refriferated or freezed, and be analyzed as soon as possible.

Objective To develop a method of support liquid-liquid extraction (SLE) and simultaneous determination of 4 components in somedon and its 8 metabolites by GC–MS/MS. Methods Somedon and its metabolites were extracted by SLE and determined by GC-MS/MS in MRM mode. The qualitative analysis was based on retention time and ratio of ions. The quantitative analysis was based on internal standard method and calibration curve. Results After SLE and determination of 4 components in somedon and its 8 metabolites, the extraction rate were 37.57%~95.87%, the linear range were 0.12μg/mL~16.00μg/mL, the correlation coefficient(r)were 0.989 6~0.999 7, LOD were 0.08ng/mL~14.48ng/mL, the accuracy were 79.63%~122.90%, the interday and intraday precision were 0.99%~7.43% and 2.19%~10.60% respectively. Conclusion Simultaneous determination of somedon and its metabolites by GC–MS/MS in biological samples, which was rapid, simple, accurate and was high precision and recovery, can be used for qualitative and quantitative analysis in forensic cases.

Objective To investigate the possibility of pulp regeneration of different irreversible pulpitis with absorbable gelatin sponge as the scaffold,and dental seed cells,induced molecules and MTA closed.Methods 150 patients with irreducible pulpitis,who were removed the crown infection pulp and retained the root living pulp,according to the visiting sequence,were randomly divided into two groups.The experimental group was given absorbable gelatin sponge as the scaffold and MTA closed.The control group was given calcium hydroxide coverage.The symptoms situation and pulp situation were observed.Results 6 months after surgery,the success rate of acute pulpitis and chronic closed pulpitis in the experimental group was 76.5％ and 47.0％,which were significantly higher than those of the control group (14.3 ％,8.8％) (P ＜ 0.01).In the experimental group,the success rate of chronic hyperplastic pulpitis and chronic ulcerative pulpitis was 33.0％ and 35.0％,which were higher than those of the control group(9.1％,6.0％),but the differences between the two groups were not statistically significant (P ＞ 0.05).Conclusion Pulp regeneration in vivo is possible with absorbable gelatin sponge as the scaffold and MTA,pulp regeneration of acute pulpitis and chronic closed pulpitis are more likely to induce.

Objective To evaluate the survival benefit of amphotericin B (AmB) plus flucytosine or fluconazole for treatment of patients with acquired immunodeficiency syndrome (AIDS)-associated cryptococcal meningitis.Methods The following database were searched from the beginning to October 2013,including Cochrane library,PubMed,OVID,Embase,Wanfang Date,CNKI and Chinese Biomedical Database,and the references of eligible studies were manually screened.Reference lists of relevant articles were screened according to selection and extraction criteria.Meta-analysis was performed using RevMan 5.2.Results Four prospective controlled studies with a total of 399 patients with cryptococcal meningitis were identified,including 386 patients with AIDS-associated cryptococcal meningitis and 13 human immunodeficiency virus (HIV)-negative patients.Two hundred and twentyseven patients were treated with AmB and flucytosine combination therapy,including 217 patients with AIDS-associated cryptococcal meningitis and 10 HIV-negative patients.One hundred and seventy-two patients were treated with AmB and fluconazole combination therapy,including 169 patients with AIDS-associated cryptococcal meningitis and 3 HIV-negative patients.The Meta-analysis revealed that the mortality rate in AmB plus flucytosine combination therapy group was 6.6％ (95％ CI:18.5％-31.6 ％) at two weeks point,which was significantly lower than that in AmB plus fluconazole combination group (19.7％,95％CI:-23.6％-62.9％; OR=0.51,95％CI:0.27-0.93,P＜0.05).But at 10 weeks point,the mortality rate in flucytosine combination group was 12.9％ (95％oo CI:-22.2％-48.0％),which was lower than that in fluconazole combination group (31.4％,95％ CI:-23.1％-85.9 ％).However,there was no statistically significant difference between these two groups at 10 weeks point (OR=0.70,95％CI:0.44-1.13,P=0.15).Conclusion Administration of AmB plus flucytosine at early stage can reduce the mortality rate in patients with AIDS-associated cryptococcal meningitis.

Objective To evaluate the clinical effect of the use of pathfile and protaper nickel-titanium instruments for machine in root canal preparation.Methods A total of 200 patients suffering from pulpitis and periapical that needs root canal treatment was randomly divided into two groups according to treatment order (100 patients/each group).The experimental group was used pathfile and protaper in root canal preparation,and filled with warm gutta vertical condensation.The control group was used protaper root canal preparatory standards method,and filled with warm gutta vertical condensation.The required time of two sets of root canal preparation and the number of equipment broken were recorded,and X-ray film at the preoperation intraoperation and postoperation were used to evaluate effect of preparation and root filling.Results In the root canals prepared of the two groups,no issued offset,no side wear,no step-forming,and no position changed of the apical foramen were occurred,which was consistent with preperative X-ray film.One case was occurred fracture of instrument in the experimental group and the control group,it was Slfile.Tooth position was right mandibular,second molar nearly the root apical 1/3,and upper left second molar mesial buccal root apical position.The root canal preparation time of experimental group was significantly shorter than the control group[the front teeth:(4.73 ± 1.12) min vs (6.32 ± 1.84) min,t =2.243,P ＜0.05; the premolars:(7.83±1.23)min vs (10.25±2.68)min,t =2.475,P ＜0.05; the molars (10.14 ± 1.18) min vs (15.43 ± 5.94) min,t =2.991,P ＜ 0.01].Postoperative X-ray showed that 2 cases were occurred small amount of paste overfilled,the root canal closed tight,no less filling; and 3 cases of the control group occurred paste overfilled,no less filling too.Conclusions The use of nickel-titanium instruments pathfile and protaper for machine in root canal preparation is feasible,and improves the efficiency of root canal preparation.

Objective To characterize clinical feature, frequency of isolation and antimicrobial susceptibility of pathogens isolated from patients with nosocomial bloodstream infections in Huashan Hospital, Fudan University from 1995 to 2004. Methods The clinical data of all patients who were diagnosed with nosocomial bloodstream infections based on national diagnostic criteria of nosocomial bloodstream infections were retrospectively analyzed. The pathogens were routinely isolated and identified. Susceptibilities against antimicrobial agents were determined by Kirby-Bauer methods and analyzed by WHONET 5.0 software. Results During the 10-year study period, a total of 395 patients were diagnosed with nosocomid bloodstream infection with 435 strains isolated from blood specimen.Gram positive bacteria, Gram negative bacilli and fungi, accounted for 47.4%, 45.1 % and 7.6%,respectively. Coagulase-negative Staphylococci (21.4%), S. aures (17.9%), E.coli (13.6%), K. pneumoniae (10.8%), Candidaspp (7.4%), Enterococci (6.0%), Pseudomonasspp (6.0%) and Acinetobacter spp (3.7%) were frequently identified isolates. S. aures and coagulase-negative Staphylococci resistant to methicillin were 62.8% and 87.1%, respectively. The susceptibilities of cefotaxime and ceftazidime against E. coli and K. preumonine were 46%-78% and 27.7%-40.4%, respectively. Conclusions The Gram positive cocci are slightly more prevalent than Gram negative bacilli in nosocomial bloodstream infections and resistance to the first line antibiotics is common among all pathogens isolated. Candida spp is the fifth leading cause of nosocomial bloodstream infections.