Objective: Proto-oncogene Zbtb7A has been characterized as a molecular switch in the process of cancer initiation and development.Our goal is to obtain the POZ domain of the Zbtb7A protein and prepare its polyclonal antibodies.Methods: We optimized the coding sequence of the POZ domain according to the codon bias of E.coli and synthesized the sequence with two-step PCR,which was then introduced into the pET-26b(+) vector to express the protein.The recombinant protein was analyzed by 15% SDS-PAGE and the corresponding band was cut out from gel.The minced gel slice that contained the POZ domain protein was injected to immunize rabbits.The collected rabbit antiserum was purified using the saturated ammonium sulfate method in combination with protein G antibody purification,and the purified polyclonal antibodies was evaluated by Western blot.Results: The optimized sequence of the POZ domain was correctly obtained and successfully constructed into the pET-26b(+) vector.After induction,an expected protein band about 14 KD was detected on 15% SDS-PAGE,and highly purified polyclonal antibodies were obtained,which were specifically bound to human Zbtb7A.Conclusion: The obtained recombinant protein of the POZ domain and its polyclonal antibodies can be used for further studies of Zbtb7A.

Objective To investigate the resident performance on preoperative anesthetist visit in Beijing hospitals,thereby providing introduction for further training.Methods A self-designed questionnaire was distributed among the anesthesia residents who were receiving anesthesia residency training in Beijing through WeChat.The questionnaire covers geographic data of the residents,information on preoperative visit and existing training program.Results160 questionnaires were reclaimed.History-taking and physical examination were not comprehensive in many residents,the nature of surgery was not evaluated by most residents.The capacity of risk assessment and risk informing were not competent in many residents.The most desired training methods for preoperative visit were scenario simulation and bedside observation.Conclusions Scenario simulation with standard patient may have a promising prospect in preoperative anesthetist visit training.

Objective:Mesenchymal stem cells( MSCs) have self-renewal capacity and potential to differentiate into the cells.It was reported that the expression of miR-30a changed in some immune diseases.But it remains unclear the effect of miR-30a on the im-munoregulatory functions of MSCs.Here we studied the impact of miR-30a on the phenotype,cell viability,apoptosis,cell cycle and im-munoregulatory functions of MSCs.Methods: The mixed enzyme methods were used for the isolation of human umbilical cord MSCs.Flow cytometry(FCM)was used to investigate the effect of overexpressed miR-30a on the phenotype of MSCs.CCK-8 was used to examine the cell viability of miR-30a-overexpressed MSCs.Annexin V/PI was used for the detection of apoptosis of MSCs.Q-PCR and Western blot were used to investigate the effect of miR-30a on the expression of Cyclin E2( CCNE2).CCNE2 was one putative target of miR-30a predicted by Targetscan database.The effects of miR-30a-overexpressed MSCs on the maturation of dendritic cells(DCs)were determined.Results:Overexpression of miR-30a blocked the cell cycle of MSCs in the G0/G1 phase by inhibiting the expression of CCNE2,but did not affect the phenotype, cell viability and apoptosis of MSCs.When co-cultured with DCs, although MSCs down-regulated the expression of CD40 and CD86 on DCs,overexpression of miR-30a more significantly enhanced the suppressive impact of MSCs on the maturation of DCs.Conclusion: miR-30a affects the cell cycle of MSCs and enhances its immunosuppressive effect on DCs.

BACKGROUND: the development of magnetic separation technique,it is feasibility to in vitro sort and amplify CD4+CD25+Treg cells for transplantation; however,the application dosage and immune tolerance have been less reported yet.OBJECTIVE: To investigate dose-effect relationship of CD4+CD25+Treg cells during allograft transplantation.METHODS: SD rats which were considered as the donors and Wistar rats as receptors were used to establish allograff kidney transplantation models.CD4+CD25+Treg cells were separated from splenic cells of Wistar rats and induced phenotype of donor antigenic specificity in vitro.According to the quantities of CD4+CD25+Treg cells injecting through tail vein during the operation of allograft kidney transplantation,models were rolled into four experiment groups: group 1(2×105),group 2(5×105),group 3(1×106),and group 4(2×106).The models out injection were considered as controls.Survival status of kidney was detected at day 15 postoperatively; creatinine level and pathological changes were detected at days 4,9 and 15 according to Banff Schema diagnostic standard; semi-quantitative scores were measured Watanabe technique.RESULTS AND CONCLUSION: The death rate was the highest in control group(83.3%),and then group 1(66.7%),group 4(58.3%),and group 2(33.3%); but rats in the group 3 were all survival.Creatinine level in experimental groups was significantly less than control group at days 4,9,and 15 postoperatively(P<0.05,P<0.01); the creatinine levels in the group1 and group 2 were significantly greater than in the group 3 and group 4 at days 9 and 15 postoperatively(P<0.05),Semi-quantitative scores demonstrated that there was no significant difference between group 2 and group 1; but the scores in the group 3 and group 4 were significantly greater than control group(P < 0.05).The results indicated that CD4+CD25+Treg cells could improve kidney function following transplantation,and prolong survival time of transplanted kidney.The 1×106 was the best dosaae for application.

Objective To investigate the effects of different doses of gabapentin on streptozotocin (STZ)-induced diabetic neuropathic pain in rats.Methods Male SD rats aged 6 weeks weighing 180-200 g were used in this study. Diabetes ntellitus ( DM) was induced by intraperitoneal STZ 60 mg/kg and confirmed one week later by blood glucose =16.7 mmol/L before breakfast. The DM rats were randomly divided into 4 groups ( n = 6 each) : gabapentin groups received intraperitoneal gabapentin 30, 60 and 120 mg/kg twice a day (at 9:00 am and 3:00 pm) for 3 weeks respectively and control group received intraperitoneal normal saline 0.6 ml instead of gabapentin. The paw withdrawal threshold to von Frey filament stimulation was measured before and at 30, 60, 120, 180, 240 min after first gabapentin injection and once a week for 3 weeks. Results After gabapentin 60 and 120 mg/kg, the paw withdrawal threshold to mechanical stimuli was significantly increased and lasted for about 4 h. The analgesic effect peaked at 60 min after IP gabapentin injection. Normal saline and gabapentin 30 mg/kg had no significant analgesic effect. The degree of analgesia was significantly decreased at day 14 and 21 of treatment with gabapentin 60 and 120 mg/kg as compared with that at 60 min after gabapentin injection. Conclusion The hyperalgesia and allodynia in rats with diabetes mellitus can be effectively reversed by gabapentin 60 and 120 mg/kg,while long-term use of gabapentin can induce drug tolerance.

Objective To observe the clinical effect and security of autohemotherapy combined with bacillus calmette-guerin polysaccharide and nucleic acid in treatment of facial steroid-dependent dermatitis.Methods The 74 patients with facial steroid-dependent dermatitis were divided into treatment group (38patients) and control group (36 patients) according to the treatment method.The patients in treatment group were treated with autohemotherapy combined with bacillus calmette-gnerin polysaccharide and nucleic acid 1 ml muscle injection,every 3 days a time for 4 weeks.The patients in control group were treated with bacillus calmette-guerin polysaccharide and nucleic acid,every 3 days a time for 4 weeks.The effect was compared between the two groups.Results The total effective rate in treatment group was 84.2％(32/38),in control group was 63.9％(23/36),there was significant difference between the two groups (P ＜ 0.05).No serious adverse reaction was found in two groups.Conclusion Autohemotherapy combined with bacillus calmettegnerin polysaccharide and nucleic acid is safe and effective in treatment of facial steroid-dependent dermatitis.

Objective The incidence rate of pressure ulcer is high in critical patients and off-loading mattresses and reposi-tioning are known as effective interventions for the prevention of pressure ulcers .However, evidence is lacking for selection of the right type of mattresses and suitable interval of repositioning .This study was to compare the effects of two types of off-loading mattresses with two different repositioning intervals in preventing pressure ulcers in critical patients . Methods According to the design of this ran-domized controlled trial , we made a training plan concerning the participants , methods of intervention and comparison , criteria and methods of observation , and methods of recording , and trained 26 nurses from 7 hospitals .Using non-inferiority design and the method of stratified blocked randomization , we divided 1194 patients with the risk of pressure ulcer into a trial group ( n=596) and a control group ( n=598) , a viscoelastic sponge mattress with every-four-hours repositioning used for the former and an automatic aeration mat-tress with every-two-hours repositioning for the latter , both for 7 successive days .We examined the patients every day , recorded the in-cidence and stages of pressure ulcer , and compared the data obtained between the two groups of patients . Results The total inci-dence rate of pressure ulcer was 1.09%(13/1194), significantly lower in the trial than in the control group (0.34%[2/596] vs 1.84%[11/598], P=0.012). Conclusion A viscoelastic sponge mattress with every-four-hours repositioning is superior to an automatic aeration mattress with every-two-hours repositioning and therefore is preferred to the latter in preventing the incidence of pressure ulcer in critical patients in the ICU .

Objective:To compare the patient-centered doctor-patient communication skills in anesthesiologists with different working experience and genders.Methods:The second and third year residents who had been trained as residents in the Department of Anesthesiology in our hospital from September 2017 and September 2018 and senior physicians who had worked for 10-15 yr in our hospital, were recruited to perform a preoperative interview in a standardized patient counter.Consultation and Relational Empathy (CARE) measure was used to assess the communication skills among senior physicians, residents, and anesthesiologists of different genders.The residents also took the objective structured clinical examination (OSCE) and theory examination.Results:Thirty-six residents and 20 senior physicians were included.There was no significant difference in the CARE measure score between residents and senior physicians and in the total CARE measure score between residents who received doctor-patient communication training at the stage of medical students and those who did not ( P>0.05). Compared with the residents in the second year, the listening ability score and decision-making ability score were significantly decreased ( P<0.05), and no significant change was found in the scores for the other items in the residents in the third year ( P>0.05). Compared with residents, the listening ability scores were significantly increased, and the caring ability scores were decreased ( P<0.05), and no significant change was found in the scores for the other items in senior physicians ( P>0.05). There was no difference in CARE measure scores between the male and female anesthesiologists.The resident′s CARE measure score was positively correlated with the annual OSCE score ( r=0.486, P<0.05), and there was no correlation between the resident′s CARE measure score and annual theoretical examination grade ( r=0.308, P>0.05). Conclusion:Senior anesthesiologists with more clinical experience are not superior to the junior residents in patient-centered communication skills, and the doctor-patient communication skills of residents are not related to their medical theoretical grade.Moreover, there is no significant difference between male and female anesthesiologists in terms of doctor-patient communication skills before surgery.Therefore, training of patient-centered doctor-patient communication skills should be strengthened at all levels of anesthesiologists.

Objective To investigate the relationship among Helicobacter pylori(H.pylori),CD4 positive cells and CD8 positive cells in gastric mucosa of the AIDS patients with gastritis.Methods Fiftyeight AIDS patients with upper abdominal pain were diagnosed with chronic gastritis through gastroscopy.The gastric biopsies from them were used for H.pylori detection with rapid urease test and Giemsa staining,pathology examination with HE staining,and immunohistochemistry analysis for CD4,CD8 positive cells in Gastric mucosa.And the application of flow cytometry was for the detection of peripheral blood CD4 and CD8 lymphocytes from the patients.Results H.pylori was positive in 26 cases,and negative was in 32 cases.CD8 cell expression in gastric mucosa of the AIDS patients with H.pylori positive was significantly higher than H.pylori negative patients(P＜0.05).There is no difference CD4 cell expression in gastric mucosa between the AIDS patients with H.pylori positive and H.pylori negative patients.Moreover,CD8 positive lymphocytes in gastric mucosa of those patients with H.pyloriinfection were significantly stronger than the CD4 positive lymphocytes.However,the peripheral blood CD4 lymphocytes from the patients with H.pylori infection were more than those from H.pylorinegative patients significantly(P＜0.05).Conclusion The expression level of CD8 cells in gastric mucosal tissues of AIDS patients with H.pylori infection were higher than those without H.pylori infection.The CD4 lymphocytes from the peripheral blood of the patients with H.pylori infection were more than those without H.pylori negative patients.

Objective To understand the current situation and existing problems in the training of healthcare-asso-ciated infection(HAI)management,and provide scientific basis for strengthening the management of HAI preven-tion and control system.Methods A questionnaire survey was adopted to investigate situation of training on HAI in 15 provincial-level HAI training agencies in China during the past 30 years,and basic condition of training on HAI management in recent 5 years.Results Among 15 provincial-level training agencies,66.67%(n=10)were respon-sible by HAI management quality control centers,80.00% have already conducted training in each city,53.33%carried out training for 10 to 20 times,33.34% performed training for ≤2 times per year.Of 33 728 trainees in 2011-2015,41.30% were 41-50 years old,61.82% were nursing staff,50.56% had bachelor degree,43.96%were with the intermediate professional title.Most trainers were HAI prevention and control experts in their respec-tive province,accounting for 68.07%,the curriculums were mainly designed on professional course,and only 26.78% were involved in management.Conclusion Professional structure of HAI management personnel is not reasonable,faculty is imbalance,knowledge update is lacking,and HAI training and education system need to be improved further.