AIM: To construct full length hCD59 eukaryotic and extracellular domain of hCD59 (hsCD59) prokaryotic expression vectors and prepare polyclonal antibody of hCD59. METHODS: cDNA fragments encoding hCD59 and hsCD59 were amplified from human PBMCs by RT-PCR and cloned into the eukaryotic vector pVAX-1 and prokaryotic vector pGEX-KG, respectively. The recombinant fusion protein GST-hsCD59 was expressed in E. coil BL21 induced by IPTG. Then the fusion protein was purified and identified. Polyclonal antibody against hCD59 was prepared by immunizing rabbit with pVAX-1-hCD59 and boosting with GSThsCD59 fusion protein, and the titer was identified. RESULTS: The recombinant eukaryotic vector pVAX-1-hCD59 and prokaryotic vector pGEX-KG-hsCD59 were successfully constructed. The GST-hsCD59 fusion protein was over-expressed in E. coli BL21 and the relative molecular mass (M~r) of the expression product was identical with predicted size. The titer of the anti-hCD59 serum was 1:3 200. CONCLUSION: We got the recombinant eukaryotic vector pVAX-1-hCD59, prokaryotic vector pGEX-KG-hsCD59 and rabbit anti-hCD59 polyclonal antibody successfully. These work would be helpful for the further study of the biological function of human CD59.

Objective To evaluate the effects of different concentrations of sevoflurane on the cognitive function in neonatal rats.Methods Twenty-four neonatal Sprague-Dawley rats of both sexes,aged 7 days,weighing 12-16g,were randomly divided into high concentration sevoflurane group (group Sev1),low concentration sevoflurane group (group Sev2) or control group (group C),with 8 rats in each group.Sev1 and Sev2 groups were exposed to 2％ and 4％ sevoflurane for 6h,respectively,and C group inhaled gas mixture composed of 30％ oxygen.At 2 and 8 weeks after exposure to sevoflurane,cognitive function was assessed using the open field test and Morris water maze test.Results In the open field test,no significant differences were observed among the three groups (P ＞ 0.05).Morris water-maze test showed that the escape latency was significantly prolonged,and the frequency of crossing the original platform and the time spent in the third quadrant were decreased at 2 weeks after exposure to sevoflurane in Sev1 group and at 2 and 8 weeks after exposure to sevoflurane in Sev2 group as compared with group C (P ＜ 0.05).Morris water-maze test showed that the escape latency was significantly prolonged,and the frequency of crossing the original platform and the time spent in the third quadrant were decreased at 2 and 8 weeks after exposure to sevoflurane in Sev2 group compared with Sev1 group (P ＜ 0.05).Conclusion Exposure to 4％ and 2％ sevoflurane for 6h both can induce cognitive decline and the decreasing effect is enhanced with the increasing concentration in the neonatal rats.

Objective:To explore the methods and ideas for developing pharmaceutical care in clinical practice. Methods: The pharmaceutical care performed by clinical pharmacists and the therapeutic scheme assisted by clinical pharmacists for one patient with adult purulent meningitis were analyzed retrospectively. Results and Conclusion:Through selection of anti-infective agents, treatment of adverse drug reactions and assessment of patients’ economic capacity,clinical pharmacists help provide reasonable medication to im-prove therapeutic efficacy, safety and economy.

Objective To investigate the effect of intrathecal injection of Tanshinone Ⅱ A on bone cancer pain behavior and spinal IL-1 β,IL-6,TNF-α expression.Methods According to the random number table method,84 C3H/HeNCrlVr male mice were divided into:(1) Tanshinone ⅡA 10 μg group:the tumor mice were treated by intrathecal administration (once daily on the days 14-20 after inoculation of tumor cells) with Tanshinone ⅡA 10 μg; (2)Tanshinone ⅡA 20 μg group:the tumor mice were treated with Tanshinone ⅡA 20 μg; (3)Tanshinone ⅡA 40 μg group:the tumor mice were treated with Tanshinone ⅡA 40 μg; (4) normal control group:the mice were given food and water ad libitum; (5) DMSO+Sham group:the sham mice were treated with the same volume of 5％DMSO; (6) Tanshinone ⅡA+Sham group:the sham mice were treated with Tanshinone ⅡA 40 μg; (7)DMSO+Tumor group:the tumor mice were treated with the same volume of 5％DMSO.The mice pain behaviors were assessed with the paw withdrawal thermal latency (PWTL) at the corresponding time points,then the mice were killed and the samples of spinal cord were detected by real-time PCR.Results The basic values of PWTL had no significant differences among all groups (P＞0.05).At day 14 after operation,no significant difference (P＞0.05) was found in the PWTL value between normal control group and the sham operation group.But in tumor group,the PWTL value was significantly lower than that of normal control group (P＜0.05).At day 21 after operation,the PWTL and the level of spinal IL-1 β,IL-6,TNF-α expression had no significant differences (P＞0.05) among normal control group,Tanshinone ⅡA+Sham group and DMSO+Sham group.The PWTL ((6.19± 1.26)s) in DMSO+ Tumor group was significantly lower than that of normal control group((16.01± 1.59)s) (P＜0.05),but the level of IL-1β,IL-6,TNF-α expression was higher than that of normal control group.Compared with the normal group,the PWTL ((9.83±1.26)s;(10.29±2.95) s) of Tanshinone Ⅱ A 20 μg and 40 μg group was higher,and the level of spinal IL-1β,IL-6,TNF-α expression was lower (P＜0.05).The PWTL and the levels of spinal IL-1β,IL-6,TNF-αexpression had no significant differences between Tanshinone ⅡA 10 μg group ((6.67 ± 0.96) s) and DMSO + Tumor group(P＞0.05).Conclusion Intrathecal injection of Tanshinone Ⅱ A plays a role in anti-cancer pain,and inhibition of spinal inflammatory cytokine release may be one of its mechanisms.

Objective To investigate the safety and effectiveness of different intravenous anesthesia methods for pediatric ERCP . Methods Data of 45 children undergoing ERCP at the Affiliated Hangzhou Hospital of Nanjing Medical University from August 2013 to July 2016, including intravenous anesthesia,the procedure of ERCP, adverse reactions and the waking time were retrospectively studied. Results A total of 45 patients in two groups under intravenous anesthesia successfully underwent ERCP . Seventeen patients ( 37. 8%) whose body weights were over 20 kg and the duration of surgery was predicted less than 30 minutes received deep sedation without airway intubation. Twenty?eight patients ( 62. 2%) with an initial weight of less than 20 kg and the duration of surgery was predicted more than 30 minutes received general anesthesia with airway intubation. In patients with deep sedation, the mean time of waking was 7. 2±6. 3 minutes, body movement reaction occurred in 1 case ( 5. 9%) and with transient decreasing of pulse blood oxygen ( beyond 95%) occurred in 2 cases ( 11. 8%) . In patients receiving endotracheal anesthesia with intubation, the mean waking time was 10. 5±8. 7 minutes without adverse reactions associated with anesthesia. Conclusion Both deep sedation and general anesthesia with airway intubation are safe for pediatric ERCP. However, general anesthesia with airway intubation is an ideal method ensuring the airway safety and oxygen supply for children less than 20 kg undergoing first?time ERCP or the duration of surgery lasting over 30 minutes.

Objective: To investigate the visual acuity of primary and middle school students in Ningxia by using decision tree model and logistic regression, and to provide evidence for the prevention and control of visual acuity. Methods: Using stratified cluster sampling method, 3 257 children aged 7-18 years were randomly selected from Yinchuan and Wuzhong in Ningxia. Visual acuity test and questionnaire survey were conducted. The C 5.0 algorithm in Clementine12.0 and Logistic regression were used to analyze the factors associated with visual acuity. Results: The overall rate of low vision of the middle school of Ningxia was 62.6%, with girls (68.1%) higher than that of boys(56.8%)(P<0.01). There was a significant difference in the prevalence of low vision among different educational stages(P<0.01), with high school students had the highest detection rate(82.2%). Results of decision tree C 5.0 algorithm showed that educational stage, gender, time to do homework after school and frequency of moderate intensity exercise per week associated with poor eyesight of primary and secondary school students. Logistic regression results showed that girls, the distance between the eyes and the computer display screen <66 cm(OR=1.24, 95%CI=1.00-1.52), and the distance between the eyes and the textbook when reading was less than one foot (OR=2.05, 95%CI=1.23-3.40) were positively associated with poor vision. Low grade students and homework time <2 hours after school (OR=0.76, 95%CI=0.62-0.92) were negatively associated with lower risk of poor vision. Conclusion Educational stage, gender and time of homework after school are associated with the visual acuity of primary and secondary school students in Ningxia. It is suggested that prevention of myopia should start from the low-grade students, reducing after-school work for primary and secondary school students, and increasing outdoor activities. All the girls should be encouraged to engage in more outdoor activities, to prevent the occurrence and development of myopia.

Objective To evaluate the modifiying efficacy of pectoral nerve ( Pecs) block com-bined with general anesthesia in patients undergoing radical mastectomy. Methods A total of 60 American Society of Anesthesiologists physical status Ⅰ or Ⅱ female patients, aged 40-64 yr, weighing 50-70 kg, scheduled for elective radical mastectomy, were divided into 2 groups ( n= 30 each) using a random num-ber table method: general anesthesia group ( group GA) and Pecs block combined with general anesthesia group ( group PB+GA) . The patients in group PB+GA received Pecs block guided by ultrasound. After the needle was advanced to the pleura plane between the pectoralis major and minor, 0. 33％ ropivacaine 15 ml was injected, and 0. 33％ ropivacaine 30 ml was injected into the surface of the serratus anterior muscle at the level of the third rib. Anesthesia was induced by intravenous injection of midazolam, propofol, sufen-tanil and cisatracurium and maintained by target-controlled infusion of propofol and intermittent intravenous boluses of sufentanil and cisatracurium. Bispectral index value was maintained at 40-60. Patient-controlled intravenous analgesia was performed with sufentanil at the end of surgery. Immediately after anesthesia in-duction, at 30 min and 2 h after skin incision, and at 2, 6 and 24 h after operation, peripheral venous blood samples were collected for determination of the concentrations of plasma epinephrine by radioimmuno-assay, and the blood samples were collected from the end of the finger at the same time for determination of blood glucose concentrations. The consumption of intraoperative sufentanil, emergence time, pressing times of patient-controlled analgesia ( PCA) and development of nausea and vomiting within 24 h after operation were recorded. Results Compared with group GA, blood glucose and plasma epinephrine concentrations were significantly decreased, the intraoperative sufentanil consumption was reduced, the emergence time was shortened, the pressing times of PCA were decreased at 2, 6 and 24 h after operation ( P<0. 05) , and no significant change was found in the incidence of postoperative nausea and vomiting in group PB+GA ( P>0. 05). Conclusion When Pecs block combined with general anesthesia is used in the patients undergoing radical mastectomy, it is helpful in reducing the opioid consumption and more helpful in inhibiting postoper-ative stress responses and pain responses than general anesthesia alone.

Clinical pharmacists joined the team for treating a cerebral hemorrhage patient with hypertension. Medications were as-sessed by learning guidelines and consulting the related literature reports,and the regimen was adjusted according to the condition of the disease and drug characteristics during the whole process of pharmaceutical care. As a result, the clinical medication became more rea-sonable. The patient's blood pressure was controlled within the target range timely, and the vital signs were sufficiently stable so that the patient could be transferred to a general ward for the further treatment.

Objective: To explore the feasibility and clinical effect of autologous fat concentrate combined with split-thickness skin grafts in the repair of venous ulcer. Methods: From January 2015 to December 2018, a total of 16 patients diagnosed with venous ulcer were admitted to our department, and all patients received symptomatic and supportive treatment such as local debridement and dressing change. After the granulation tissue grew well, all patients received autologous adipose concentrate combined with spilit-thickness skin graft for repair. Results: All the grafted skin survived well, and the survival rate of grafted skin at 2 weeks after surgery was 86% to 99%. All patients were followed up to 6 months after surgery. The wounds of all patients were healed during the follow-up period. One patient suffered local ulcer recurrence due to repeated friction after healing and the ulcer recovered after dressing changing treatment. All the other patients had no recurrence during the follow-up period. The scar hyperplasia in the skin transplantation area was not obvious and the healing quality was good. No significant surgical complications occurred. Conclusions Autologous lipoconcentrate combined with split-thickness skin grafts is an effective and safe method in the repair of venous ulcer, and the application of autologous lipoconcentrate can effectively improve the survival rate and healing quality of skin grafts.

Objective To determine the median effective dose (ED50) of oxycodone inhibiting responses to laryngeal mask airway (LMA) insertion when combined with propofol in the adult female patients.Methods Female patients,aged 18-60 yr,with body mass index of 18-24 kg/m2,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,scheduled for elective gynecologic surgery,were included.ED50 of oxycodone was determined by up-and-down sequential technique.Anesthesia was induced by target-controlled infusion of propofol at target plasma concentration of 3.0 μg/ml.Oxycodone was injected at the initial dose of 0.11 mg/kg after the target effect-site and plasma concentrations were balanced.LMA was inserted at 5 min following injection.When the response to LMA insertion was positive,the concentration of oxycodone was increased/decreased in the next patient.The difference between the two successive doses was 0.02 mg/kg.LMA insertion response was defined as occurrence of swallowing,biting on the LMA and bucking and/or body movement during insertion.Probit analysis was used to calculate the ED50 and 95％ confidence interval of oxycodone inhibiting responses to LMA insertion when combined with propofol.Results The ED50 (95％ confidence interval) of oxycodone inhibiting responses to LMA insertion was 0.095 (0.080-0.108) mg/kg when combined with propofol.Conclusion The ED50 of oxycodone inhibiting responses to LMA insertion is 0.095 mg/kg when combined with propofol in the adult female patients.