The effect of methanol addition on the heterologous expression of isoprenyl transferase NovQ was studied in Pichia pastoris Gpn12, with menadione and isopentenol as precursors to catalyze vitamin K2 (MK-3) synthesis. The expression of NovQ increased by 36% when 2% methanol was added every 24 h. The influence of initial pH, temperature, methanol addition, precursors (menadione, isopentenol) addition, catalytic time and cetyltrimethyl-ammonium bromide (CTAB) addition were explored in the P. pastoris whole-cell catalytic synthesis process of MK-3 in shaking flask. Three significant factors were then studied by response surface method. The optimal catalytic conditions obtained were as follows: catalytic temperature 31.56 ℃, menadione 295.54 mg/L, catalytic time 15.87 h. Consistent with the response surface prediction results, the optimized yield of MK-3 reached 98.47 mg/L in shaking flask, 35% higher than that of the control group. On this basis, the production in a 30-L fermenter reached 189.67 mg/L when the cell catalyst of 220 g/L (dry weight) was used to catalyze the synthesis for 24 h. This method laid the foundation for the large-scale production of MK-3 by P. pastoris Gpn12.

Objective To compare the early clinical efficacy of renal transplantation between extended criteria donor (ECD) and standard criteria donor (SCD). Methods Clinical data of 85 recipients undergoing renal transplantation from donation after cardiac death (DCD) were retrospectively analyzed. According to the types of donors, all recipients were divided into the ECD group (n=31) and SCD group (n=54). The level of serum creatinine (Scr), incidence of early complications and clinical prognosis within 3 months after renal transplantation were compared between 2 groups. Results No statistical significance was observed in the levels of Scr within 1 month after renal transplantation between the ECD group and SCD group (all P>0.05). At postoperative 60 and 90 d, the level of Scr in the ECD group was (189±97) and (175± 69) μmol/L respectively, significantly higher than (142±49) and (135±41) μmol/L in the SCD group (P=0.005 and 0.002). In the ECD group and SCD group, the incidence of acute rejection (AR) was 6% and 15%, the incidence of delayed graft function (DGF) was 23% and 19%, the incidence of pulmonary infection was 10% and 6%, the incidence of other early complications was 32% and 15%, respectively, no statistical significance was identified (all P>0.05). In the ECD group and SCD group, the survival rate of the recipient was 97% and 94%, the survival rate of the renal was 84% and 91%, no statistical significance was identified (all P>0.05). Conclusions Compared with the SCD, renal transplantation from ECD can achieve equivalent early clinical efficacy. In the present condition of serious deficiency of donor kidney, the application of ECD can enlarge the supply of the donor kidney.

Objective To observe the clinical efficacy and prognosis of nimotuzumab combined with intensity modulated radiotherapy(IMRT) in the treatment of advanced local nasopharyngeal carcinoma.Methods 82 patients with advanced nasopharyngeal carcinoma were selected, and they were randomly divided into observation group(40 cases) and control group(42 cases) according to the digital table.The observation group was treated with nimotuzumab plus IMRT plus cisplatin, and the control group was treated with IMRT plus cisplatin.After 7 weeks of treatment, the clinical efficacy, adverse reactions and T cell subsets were observed.And the patients were followed up for 1 year, 2 years and 3 years to evaluate the survival rate.Results In the observation group, the total remission rate was 75.00%,which was higher than that in the control group(59.09%),the difference was statistically significant(x2=35.81,P<0.05).The CD+4,CD+8 and CD+4/CD+8 levels in the observation group were similar to those in the control group(tCD+4=0.59,P=0.56;tCD+8=1.70,P=0.01;tCD+4/CD+8=0.13,P=0.89).The incidence rates of Ⅰ degree and Ⅱ degree mucosal reaction and dermatitis in the observation group were 35.00%,40.00%,40.00%,35.00%,respectively,which were higher than 9.09%,9.09%,9.09%,9.09% of the control group,the differences were statistically significant(x2Ⅰmucous membrane=4.18,P=0.04;x2Ⅱmucous membrane=5.52,P=0.02;x2Ⅰdermatitis=5.52,P=0.02;x2Ⅱdermatitis=4.18,P=0.04).The 2-year and 3-year survival rates in the observation group were 65.50%,60.00%,respectively,which were significantly higher than 31.82%,27.27% in the control group, the differences were statistically significant(x2=4.11,P=0.04;x2=4.58,P=0.03).Conclusion Nimotuzumab combined with IMRT in the treatment of advanced nasopharyngeal carcinoma has good short-term clinical efficacy, good long-term efficacy and controllable adverse reactions.It is worthy of clinical application.

The service capabilities and sustainable development of community doctors' first contact are undoubtedly the critical factors to promote the construction of grassroots clinics and tiered health care system.Based on the review of motivation-related concepts and its development, as well as the analysis of the current conditions of community first contact and characteristics of community doctors in China, this paper puts forward and defines the concept and connotation of service motivation for community doctors' first contact by logically summarizing the concept of force in physical sciences ,motivation in service science, Based on analyzing the extrinsic motivations to form community doctors' first contact such as attraction, support, constraint, and stress the intrinsic motivations produced by professional identity, self-efficacy and achievement,as well as the interactions and relationships between them, this paper contructs a formation mechanism of the service motivation for community doctors' first contact, which is visual and intuitive.This paper provides a logical path and basis for scientific governance and upgrade of the service motivation for community doctors's first contact to effectively promote community first contact system in China.

Objective To evaluate the effect of hypothermia status in the donors upon the renal graft function after renal transplantation from donation after citizen's death. Methods Thirty-six eligible donors were randomly divided into the normal temperature (body temperature 36.5-37.5 ℃ , n=19) and hypothermia groups (body temperature 34.0-35.0 ℃ , n=17). The matched recipients undergoing renal transplantation were also assigned into the normal temperature (n=38) and hypothermia groups (n=34). Perioperative conditions of the donors and recipients were compared between two groups. And postoperative renal graft function of the recipients were statistically compared between two groups, including the incidence of delayed graft function (DGF) and primary nonfunction (PNF). Results No statistical significance was identified in the perioperative amount of urine volume, serum creatinine (Scr), systolic blood pressure, saturation oxygen, warm ischemia time and cold ischemia time of the donors between two groups (all P>0.05). No statistical significance was noted in terms of the operation time, intraoperative mean blood glucose and intraoperative mean arterial pressure of the recipients between two groups (all P>0.05). Postoperative incidence of DGF of the recipients in the hypothermia group was 6%, significantly lower than that in the normal temperature group (24%) (χ2=4.393, P=0.036). Postoperative incidence of PNF of the recipients was 3% in both the hypothermia and normal temperature groups with no statistical significance (χ2=0.000, P=1). Conclusions The hypothermia status of the donors can significantly reduce the incidence of DGF, whereas exerts no evident effect upon the incidence of PNF in the recipients.

Objective To investigate the clinical characteristics, prevention and treatment of multi-drug resistant organisms (MDROs) infection early after renal transplantation from donation after citizen's death. Methods Clinical data of 166 patients undergoing allogeneic renal transplantation and regular follow-up in Xijing Hospital from November 2011 to September 2016 were retrospectively analyzed. General conditions were statistically compared between the recipients undergoing renal transplantation from donation after cardiac death (DCD) and their counterparts receiving living related donor renal transplantation. The incidence of MDROs infection, onset time, course of diseases, complications, infection site and etiological type were observed. The therapeutic methods and clinical prognosis were summarized. Results The incidence of MDROs infection early after renal transplantation in the recipients undergoing DCD renal transplantation was 14%, significantly higher than 2% in those receiving living related donor renal transplantation, and 13% and 2% for the incidence of delayed graft function with statistical significance (both P<0.05). The incidence of renal graft loss was 8%and 2%, and 5% and 1% for the mortality rate without statistical significance between two groups (both P>0.05). MDROs infection occurred in 11 patients after DCD renal transplantation. The most common infection site was urinary system(n=6) and the most prevalent pathogenic bacterium was Escherichia coli (n=4). All patients infected with MDROs were treated with a sufficient dosage of effective antibiotics according to the outcomes of bacterial culture and drug sensitivity test. Eight patients obtained favorable clinical prognosis, one underwent nephrectomy and two died. Conclusions The incidence of MDROs infection early after DCD renal transplantation is higher than that after living related-donor renal transplantation. Strict donor screening, early detection, intimate monitoring and timely treatment can effectively reduce the risk of MDROs and enhance clinical prognosis.

Objective To investigate the effects of thyroid hormone on extremely severe traumatic brain injury (TBI).Methods A retrospective case-control study was conducted to analyze the treatment of 105 patients with extremely severe TBI admitted from July 2010 to April 2014.There were 79 males and 26 females,with an average age of 32.9 years.The patients were divided into conventional treatment group (Group A,35 cases),conventional treatment ± thyroxine treatment group (Group B,35 cases) and thyroxine treatment group after the condition that thyroxine level was low (Group C,35 cases) according to the random number table method.The incidence of low T3 and T4,incidence of hypotension,the dosage of vasoactive drugs,function evaluation of liver and kidney damage,Glasgow outcome scale (GOS),and acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) within 20 days after admission,and mortality rate within 30 days after admission were compared and analyzed.Results Within 20 days after admission,the rates of low thyroxine levels and hypotension of the Group B (22.9％,77.1％) were significantly lower than those in the other two groups (Group A:40％,100％;Group C:37％,100％) (all P ＜ 0.05).The doses of dopamine and norepinephrine in Group B was significantly lower than the other two groups and the combination rate of vasopressors in Group B was significantly lower than the other two groups (P ＜ 0.05),while there was no significant difference between Group A and Group C (P ＞ 0.05).The corresponding data in Group A and Group C had no statistically significant difference (P ＞ 0.05).The liver and renal dysfunction rates of Group B (29％,31％) were significantly lower than those of the other two groups (Group A:49％,51％;Group C:43％,51％) (all P ＜ 0.05).The corresponding data in Group A and Group C had no statistically significant difference (P ＞ 0.05).GOS in Group B [(4.8 ± 1.9) points] was significantly higher than that in Group A [(3.3 ± 0.2) points] (all P ＜ 0.05) within 30 days after admission and significantly higher than that of itself at the beginning [(3.6 ± 1.1) points] (P ＜ 0.05).The APACHE Ⅱ in Group A was significantly higher than those in other two groups as well as that in Group A at admission (P ＜ 0.05).Mortality rates in Group B (31％) and Group C (29％) were significantly lower than that in Group A (69％) within 30 days after admission (P ＜ 0.05).Conclusions Thyroxine can reduce the incidence of hypotension,liver and kidney injury rate in extremely severe TBI.Prevention is better than the supplementary treatment after severe TBI.Thyroxine can also reduce the mortality of extremely severe TBI within 30 days after admission.

Objective To study the effect of ING5 gene on growth inhibition of gastric cancer. Methods ING5 expressing plasmid was transfect?ed into SGC?7901 cells. The cell viability was assessed by CCK?8,cell apoptosis and cycle was detected by flow cytometry analysis,the migration and invasion was evaluated by scratch and transwell,the expressions of mRNA and protein were determined by real?time quantitative PCR and West?ern blot respectively. Nude mice were implanted subcutaneously with SGC?7901 cells and tumor size and related protein were analyzed. Results After transfection of pEGFP?N1?ING5,the proliferation of gastric cancer SGC?7901 cells was significantly inhibited(P<0.05),the apoptosis was decreased(P<0.05),the percentage of G1 phase was increased and G2 phase was decreased(P<0.05),the ability of migration and invasion was reduced(P<0.05),the expression of NF?κB,PI3K,p?Akt1/2/3,Bcl?2,XIAP,β?catenin,c?myc mRNA,and protein levels were significantly in?creased(P<0.05),and the expression of Akt1/2/3、MMP9 mRNA and protein levels were significantly decreased(P<0.05). Conclusion The ING5gene inhibits the SGC?7901 cell proliferation,induces G1 arrest,inhibits the migration and invasion,and effectively regulates the related genes and proteins about cell proliferation,cell cycle and adhesion,but reduces apoptosis. ING5 can inhibit the evolution and development of gastric can?cer.

Objective To explore the effects and molecular mechanisms of suberoylanilide hydroxamic acid (SAHA) on ovarian carcinoma. Methods (1)Two groups of ovarian carcinoma cell lines (SKOV3 and SKOV3/DDP, HO8910 and HO8910-PM) were exposed to SAHA (1, 3, 5 and 7μmol/L SAHA,group 1-group 4). CCK-8 method was employed to eval-uate the inhibitory effects of SAHA.(2)Ovarian cancer cell lines treated with SAHA (2 or 5μmol/L SAHA) were used as 1 and 2 groups. Flow cytometry was performed following staining with Annexin V-FITC and PI for cell cycle and apoptosis.(3) Reverse transcription polymerase chain reaction (RT-PCR) and Western blot assay were used to assess the mRNA and pro-tein expression levels of phenotypic correlation factor. Results (1)After 48 h of SAHA treatment,the OD value of SKOV3, SKOV3/DDP,and HO8910 showed a trend of gradually reduce (P<0.05).(2)The apoptotic rates were significantly higher in SAHA 1 and SAHA 2 groups than those of control group (P<0.05). Compared with control group, after 48 h of SAHA treat-ment,S phase and G2/M phase of SKOV3 and SKOV3/DDP cells increased;G0/G1 phase of HO8910 and HO8910-PM cells increased in SAHA 1 and 2 groups (P<0.05).(3)The expression levels of CyclinB1 and Cdc2 (p34) mRNA were significant-ly lower in SAHA 1 and 2 groups than those of control group,while the expression levels of Caspase-3,p21 and p53 mRNA expression were significantly higher in SAHA 1 and 2 groups than those of control group. Furthermore,the expression of Ac-Histone H3,Ac-Histone H4,p53 protein were markedly improved,and CyclinB1,Cdc2(p34) protein decreased in SAHA 1-4 groups. Conclusion SAHA may suppress cell growth, induce apoptosis and cause cycle arrest in ovarian carcinoma cells by promoting histone acetylation or modulating their phenotype-related proteins of Caspase-3, p53, CyclinB1 and Cdc2(p34).

Objective To investigate the effect of anisodamine on oxygen metabolism and organ function in dogs with sepsis.Methods Twenty beagles were divided into sepsis group (n =8) and anisodamine-treated group (n =12) according to the random number table.The sepsis model was established by cecal ligation and puncture.Changes in thrombomodulin (TM),arterial oxygen partial pressure (PaO2),oxgen uptake rate (ERO2) and lactate were mearsured before and 1,3,8 and 24 h after operation.Wet/dry ratio of lung tissues and incidence of multiorgan dysfunction syndrome (MODS) were detected 24 h after operation.Results Level of TM was increased progressively after operation,which was significantly higher in sepsis group than in anisodamine-treated group at 3,8 and 24 h [(28.7±15.9) vs (19.5 ±10.6)ng/ml,(36.5±20.8) vs (21.9±12.8)ng/ml,(52.6±32.7)vs (31.3 ± 18.4)ng/ml respectively] (P ＜ 0.05).The PaO2 decreased slowly after operation,with the decrease more significant in sepsis group than in anisodamine-treated group at 24 h [(65.6 ± 14.5) vs (83.3 ± 16.3)mmHg] (P ＜0.05).The ERO2 had a rise-and-fall trend after operation,with the level lower in sepsis group than in anisodamine-treated group at 24 h [(16.7 ± 3.1) ％ vs (21.7 ± 2.9) ％] (P ＜ 0.05).Level of blood lactate increased gradually,with the level higher in sepsis group than in anisodamine-treated group at 24 h [(4.4 ± 1.2) vs (3.1 ± 0.8) mmol/L] (P ＜ 0.05).Wet/dry ratio of lung tissues in sepsis group was (510 ± 80) ％ versus (370 ± 50) ％ in anisodamine-treated group at 24 h (P ＜ 0.05).Incidence of MODS in sepsis group was 38％ versus 17％ in anisodamine-treated group at 24 h (P ＜ 0.05).Conclusion Anisodamine may ameliorate the injury to vascular endothelial cells and oxygen metabolism disorder after sepsis and hence contributes to the reduction of sepsis-induced MODS.