Objective:To construct an immune inhibitor of new TNF-?,C-terminal sequence of new rhTNF-? was replaced with sequence of T-help cell of hen egg-white lysozyme(HEL).Methods:The rhTNF-? mutant was cloned 、expressed and purified.Results:The DNA sequencing analysis showed that the C-terminal sequence of new rhTNF-? mutant was correct.The mutant was inserted into pBV220 expression vector .After the recombinant bacteria was incubated at 42℃ for 4 h,a new band of the protein with relative molecular weight of 1.7 kD was shown on the gel.The band amounted to 30 % of total bacteria protein.Western blot showed that the mutant protein could associated with anti-TNF-? antibody.After the protein was purified by through a column of Q-Sepharose Fast Flow, the purity of the protein was above 90%.The biological activity of the protein was measured with L929 cells.The result showed that biological activity of the protein was totally lost.Conclusion:The experimental evidence demonstrated that the construction of new rhTNF-? mutant was successful.The mutant not only can be associated with anti-TNF-? antibody, but also lose the biological activity of original TNF-?. [

Child ; Drug Resistance, Viral ; HIV ; drug effects ; genetics ; HIV Infections ; virology ; Humans

Objective:To compare the pain control efficiency of continuous adductor canal block (ACB) and femoral nerve block (FNB) in total knee arthroplasty.Methods:From April to September 2016,patients with severe knee osteoarthritis undergoing primary unilateral total knee arthroplasty (TKA) were prospectively observed,and all the patients were randomized received ultrasound-guided continuous ACB or FNB after surgery.Numeric pain rating scales (NPRS)pain scores in rest and activity 2,6,12,24 and 48 h after surgery were collected,and the preoperative and postoperative quadriceps strength at 24 and 48 h were analyzed.Opioids consumption and anesthesia related adverse effects were also recorded.Results:In the study,40 patients were enrolled,with 20 patients in each group,male ∶female =7 ∶ 33,the age:(63.8 ± 10.1) years,and the body mass index (BMI):(28.5 ± 3.5) kg/ m2.The general conditions were comparable between the two groups.Though the rest pain 2 h after surgery [ACB =0.0(0,6),FNB =3.0(0,5),P=0.004] and activity pain 12 h post operation [ACB =3.0(3,0),FNB =5.5(0,10),P =0.004] were lower in ACB group compared with FNB group,there was no statistical difference in the other pain checking points between the two groups.The quadriceps strength 24 h and 48 h after surgery were (85.3-± 27.6) N and (80.0 ± 30.1) N in ACB group,(69.0 ± 29.4) N and (64.4 ± 32.0) N in FNB group,both of them were declined by time.The exact data were higher in ACB group,however,there was no statistical difference between the two group by repeated measurements variance analysis(F =2.703,P =0.108).Four patients in ACB group and five in FNB acquired additional use of dolantin once (100 mg/per time) within 24 h.And among them,three patients acquired once dolantin in ACB,two in FNB,from 24 to 48 h postoperation.There were five patients who suffered nausea postoperation in ACB group,and one who reported xerostomia.Four patients in FNB had nausea with vomiting,and three experienced xerostomia.Deep vein thrombosis appeared in 2 patients in FNB group,but no one in ACB group.Conclusion:Continuous ACB is not superior in pain control after TKA compared with FNB,and the quadriceps strength could be reserved more by this method,which performed early benefits in fast rehabilitation.

Objective To modify the method of gavage administration in guinea pigs. Methods Fourty awake guinea pigs were kept rearing on the hind legs and leaning on a vertical fixture to avoid their escaping forward. A 1 mL injector was inserted into the mouth to the depth when the molar teeth were passed. Another fourty guinea pigs under general anesthesia were reversed at trendelenburg position and a children suction tube with an outer diameter of 2 mm was inserted into the stomach. Results All of the 80 guinea pigs were administered by modified gavage smoothly for seven consecutive days by one operator each time. None endured much pain or digestive tract injury, or died from air way perfusion by mistake. Conclusions We successfully modified the gavage method in guinea pigs, which would definitely take guinea pigs involved in intragastical pharmacal experiments besides the routine of rats and mice.

BACKGROUND: Dynamic hip screw (DHS) is a standard internal fixation for intertrochanteric fracture, whereas the patient combined with osteoporosis, cut-out incidence of lag screw is common. The articles in China and abroad indicate bone cement augmentation of DHS to achieve firm fixation. As for normal bone, no reports is published that whether bone cement augmentation is effective.OBJECTIVE: To investigate the biomechanics of DHS with bone cement augmentation for the fixation of intertrochanteric fracture specimen that has a normal bone density.DESIGN, TIME AND SETTING: Bilateral contrast observation study of the same sample was performed in the Laboratory of Biomechanics, Hebei Orthopaedic Research Institute (Shijiazhuang, Hebei Province, China) between March and April In 2005.MATERIALS: Bilateral upper femora from the embalmed male cadaver were provided by Anatomy Department of Hebei Medical University (China). X-ray scan results proved the absence of tuberculosis, anatomical deformity and tumor.METHODS: Twenty-four matched pairs of the upper femora (48 sides) were used to make the specimens of the intertrochanteric fracture of type A2. The right femur specimens were fixed with DHS augmented by bone cement, as the augmentation group (The screw track of femoral neck was expended by curette, and the femoral head facing upwards were injected with 2mL low viscosity bone cement. Then lag screw was wrested to keep the position unchanged till the bone cement coagulated. Placing barrel, compressing through tighten tail screw, and cortical screw fixing side-plate were. followed). And the left femur specimen was fixed with DHS conventional fixation, as the control group. The bending and torsional tests were performed in the two groups.MAIN OUTCOME MEASURES: The maximum load and the maximum torque in the two groupsRESULTS: The maximum load and the maximum torque were (3852.1602±143.6031) N and (15.5+2.6) Nm in the augmentation group, and (3702.9667±133.8601) N and (14.7±3.4) Nm in the control group. There was no significant difference in the biomechanical effects between the two groups (P>0.05).CONCLUSION: The augmenting fixation with bone cement for intertrochanteric fracture specimen has no significant effect on the strength of DHS fixation or on the overall stability of the fractured bone in normal bone density.

Objective To build the cohort of drug resistance and analyze treatment efficiency of AIDS patients and situation of drug resistant mutations among HIV-1 infected individuals.Methods A cohort of 116 HIV-1 infected patients was built and their treatment progress were acquired once every 6 months.At the sanle time CD4+ T cell counts and HIV-1 viral load were measured and genotyping for drug resistance was determined by a home brew nested PCR.Results The CD4+ T cell count(470±251/ml)was higher than that before treatment in patients who were treated by AZT/DDI/NVP or D4T/DDL/NVP.The viral load was lower than that before treatmenL The drug resistant mutation frequency increased gradually along with treatment.The CD4+ T cell count was decreased and viral load was increased and the prevalence of drug resistant mutation was increased in the patients who changed regimens to AZT/3TC/NVP or D41/3TC/NVP.Only one primary mutation that was resistant to non-nucleoside reverse transcriptase inhibitors (NNRTIs)was detected in the naive patients.The cross-resistant mutation was detected in two patients after 6 months treatment. The intermediate resistance to lopinavir(LPV) was detected after 12 months treatment.The prevalence of high-grade resistances to NNRTIs was increased obviously,and the prevalence of multi-resistance and cross-resistance was detected in 5 patients after 36 months treatment.Conclusions The prevalence of primary mutation was rare in naive HIV-1 infected patients.The prevalence of drug resistant mutation was inereased gradually along with treatment.Ahhough few regimens were available,the treatment effect could last relatively long period of time if patients keep taking medicine stably.The regimens could be changed according to the results of drug resistant test.

Objective To research the incidence and etiological factors of intraocular pressure(IOP)elevation in patients in early and late postop?erative stages after vitrectomy. Methods The clinical data of 235 cases(306 eyes)who underwent vitrectomy in our hospital were respectively ana?lyzed. IOP was measured before operation,in the early postoperative stage(within 2 weeks)and in the late postoperative stage(6 months after opera?tion or longer)by non?contact tonometer. Risk factors related with ocular hypertension happened in the early and late postoperative stages were statis?tically analyzed. Results The mean average IOP before operation was 15.3 ± 4.1 mmHg for the 306 eyes. Within the follow?up period of 6?20 months(mean,14.3 months),ocular hypertension occurred in 42 eyes[incidence,13.7%;mean,31.9 ± 6.0 mmHg]in the early postoperative stage and in 12 eyes[incidence,3.9%;mean,32.1 ± 5.7 mmHg]in the late postoperative stage. Logistic regression analysis revealed that lensecto?my,scleral buckling,cyclophotocoagulation,inert gas filling,silicone oil tamponade and diabetic retinopathy were the independent risk factors of oc?ular hypertension in early postoperative stage after vitrectomy,while lensectomy,scleral buckling,pan retinal photocoagulation,silicone oil tampon?ade,diabetic retinopathy and carotid artery stenosis were the independent risk factors of ocular hypertension in late postoperative stage after vitrecto?my. In addition,cyclophotocoagulation was a protective factor of IOP in the late postoperative stage. Conclusion The risk factors of ocular hyperten?sion after vitrectomy include the intraocular gapfiller,the mode of combined operation,disease type and carotid artery stenosis. IOP elevation that happens in the late postoperative stage is more harmful. Ocular hypertension in the late postoperative stage is not associated with that in the early stage.

Objective To evaluate the diagnostic and therapeutic effect of endoscopy on pharyngeal papillomas. Methods Data of patients with pharyngeal papillomas diagnosed and treated by endoscopy be?tween March 2009 and December 2014 were analyzed retrospectively, including dissection, treatment and follow?up results. Results The rate of endoscopic diagnosis of pharyngeal papillomas was 0?9%( 65/6 927) . Endoscopic biopsy forceps resection was performed successfully in 54 patients. Other patients ( n=11) were treated by endoscopic snare resection. There was hemorrhage of different degrees after resec?tion. Argon hemostasis was used in 6 patients for errhysis after resection. Supportive treatment was not given and no severe hemorrhage or death was seen. A total of 41 patients were followed up and endoscopic examina?tion was performed 2 months later. Pharyngeal papillomas were found again at the same site in two patients and endoscopic biopsy forceps resection was performed. Pharyngeal papillomas were not found in these two patients in the next endoscopic examination. Conclusion Endoscopy is a safe and effective diagnosis and treatment method for pharyngeal papillomas.

Objective To compare the applicability of the Beijing Version of the Montreal Cognitive Assessment (MoCA) and the Mini Mental State Examination (MMSE) in screening for cognitive impairment in patients with acute ischemic stroke for 2-3 weeks.Methods MoCA and MMSE were conducted in 201 patients with acute ischemic stroke within 2 to 3 weeks after the onset of stroke.With MoCA＜23 and MMSE ＜26 as the cut off value,we assessed the clinic effect of the MoCA and MMSE and explored the correlation between two instruments.Results The average scores of MoCA and MMSE scale were (20.5±4.3) and (25.4±3.5) points.The prevalence of cognitive impairment evaluated with MoCA and MMSE were 57.2％and 43.3％,respectively.MoCA showed significant correlation with MMSE score (Pearson's correlation coefficient=0.833,P＜0.001),and an agreement with Kappa values of 0.532 (P＜0.01) in screening for cognitive impairment.Conclusions The prevalence of cognitive impairment assessed with MoCA is higher than that of with MMSE when using MoCA＜23 and MMSE＜26 as the cut off values.Both instruments show a good agreement for screening cognitive impairment in acute ischemic stroke within 2 to 3 weeks following the disease onset.

Objective To investigate the clinical effect of pulmonary rehabilitation therapy including respiratory exercise and vibration expectoration on patients with pulmonary infection after abdominal surgery.Methods A retrospective case control study was conducted.Seventy-six patients with pulmonary infection after abdominal surgery admitted to the First Affiliated Hospital of Hunan Normal University from September 2015 to September 2016 were enrolled.According to whether accept the pulmonary rehabilitation therapy or not,the patients were divided into two groups.In the control group (n =35),the convemional expectoration method was adopted.The patients in pulmonary rehabilitation group (n =41) received both methods of the control group and pulmonary rehabilitation treatment,including respiratory exercise (effective cough,lip reduction breathing),respiratory exercise device (respiratory exerciser tri-ball),and vibrated expectoration.The 24-hour sputum volume,degree of comfort,inflammatory and pulmonary function parameters,and recovery situation were recorded in the two groups.Results ① There were no significant differences in the parameters of inflammation and pulmonary function before treatment between the two groups.After treatment,the white blood cell (WBC) and C-reactive protein (CRP) in both groups were significantly decreased,and the forced expiratory volume in 1 second (FEV1) and FEV1/forced vital capacity (FVC) were significantly increased.The above changes in pulmonary rehabilitation group were more significant than those of the control group [WBC (× 109/L):11.12 ± 2.88 vs.13.42 ± 2.62 at 3 days,8.22 ± 1.48 vs.9.27 ± 1.92 at 5 days;CRP (mg/L):13.47 ± 4.77vs.16.03±4.94 at 3 days,9.69±1.56 vs.11.77±1.41 at 5 days;FEV1 (L):2.48±0.14 vs.2.29±0.16 at 3 days,FEV1/FVC:0.78±0.04 vs.0.75±0.04 at 3 days;all P ＜ 0.05].② The 24-hour sputum volume within 3 days of pulmonary rehabilitation group were significantly higher than that of the control group (mL:30.51 ± 4.15 vs.18.30 ± 3.64at 1 day,31.08±3.22 vs.20.37±3.20 at 2 days,29.03±2.55 vs.19.03±2.51 at 3 days,all P ＜ 0.01].③ In the pulmonary rehabilitation group,the recovery time of pulmonary infection symptoms (days:5.44 ± 1.45 vs.6.20 ± 1.55),the days of antibiotic use (days:12.61 ± 3.15 vs.15.03 ± 3.78),the time of getting out of the bed (days:4.05 ± 0.74vs.4.51±0.89),and the hospital days (days:19.95±3.90 vs.22.00±4.42) were significantly shorter than those of the control group (all P ＜ 0.05),and the degree of comfort was significantly better than that of the control group (comfort score:2.71 ±0.90 vs.2.14±0.91,P ＜ 0.01).Conclusion The application of pulmonary rehabilitation including respiratory exercise and vibration expectoration in abdominal surgery patients with pulmonary infection can promote recovery,and it has a good clinical and practical application value.