Objectives:To develop a preparative method of continuous acid-urea-polyacrylamide gel electrophoresis (CAU-PAGE) to purify the defensin components from human and rabbit neutrophils (HNP-1～3 and RNP-1～5).Methods:Neutrophils were isolated from human peripheral blood or rabbit artificial aseptic ascites.The components of defensin were extracted by 5% acetic acid directly followed by elution of CAU-PAGE,and purified HNP-1～3 or RNP-1～5 were harvested.Polyacrylamide gel filtration was used as the method for comparison.Results:For each elution circa 1.50 mg of purified HNP or 2.50 mg of RNP were obtained from 5×108 of neutrophils,and their natural biological activity was preserved.Conclusions：CAU-PAGE is a relatively simple,rapid,economical method for purifying cationic peptides as the components of defensin.

Objective To study the liver X receptor(LXR)and the gene expression of cholesterol ester transfer protein(CETP),cyclooxygenase-2(COX2)in obstructive sleep apnea hypopnea syndrome(OSAHS)patients.Methods Macrophages were separated from the blood drawn in 30 cases of OSAHS patients and 30 healthy control subjects in the morning,RT-PCR gel eletrophoresis was used to determine the patterns of gene expression.Results OSAHS patients had higher LXR,COX2,CETP expression(P

Objective To determine the anti-SARS coronavirus (SARS-CoV) activity of defensins from human and rabbit, and explore the effects of defensin during the onset and progression of SARS. Methods Defensins were purified from neutrophil of human and rabbit respectively, namely HNP (human neutrophil protein) 1-3 and RNP (rabbit neutrophil protein) 1-5, by acid-urea-polyacrylamide gel electrophoresis (CAU-PAGE). The effect of defensin on Vero-E6 cell infected by SARS CoV BJ01 strain as well as the replication of virus were observed by micro-culture technique. Rusults The HNP 1-3 and RNP 1-5 separated by CAU-PAGE presented a single band with molecular weight of approximately 4000 dalton on 15% SDS-PAGE. HNP 1-3 was identified and validated by quantitative ELISA. The activity of anti-escherichia coli (ATCC25922 strain) of HNP 1-3 and RNP 1-5 was tested by MIC determination method, and the IC50 values obtained were 25～50?g/ml and 5～10?g/ml, respectively. In the antiviral experiment, HNP 1-3 and RNP 1-5 in the concentration from 25 to 100?g/ml were not able to prohibit cytopathic effect (CPE) of SARS-CoV and the replication of virus in Vero-E6. Conclution The natural defensins (HNP 1-3 and RNP 1-5) purified from neutrophils do not play any prohibitory or eliminative effects on SARS-CoV, indicating that that human and rabbit do not have natural immunity against SARS-CoV.

A brief description of performance appraisal as used in the healthcare sector, and an analysis of problems existing in performance appraisal for posts in community heahhcare organizations in the country. The paper proposes a job-analysis based performance appraisal for such posts, featuring the appraisal model of "performance + skills + attitude+ work intensity". On the basis of work analysis, a job description should be worked out for such healthcare workers as doctors, nurses and prevention workers in community healthcare organizations, aided by a targeted job-based performance appraisal system. These efforts can effectively enhance the job performance in such organizations, catering to the healthcare needs of community residents.

AIM:To explore the regulation of lumbar physiological curvature by angulation traction and longitudinal rotation and the clinical significance of repairing spinal balance on lumbar disc herniation(LDH). METHODS:①Totally 71 patients with LDH diagnosed by CT at the 451 Hospital of Chinese PLA were enrolled from October 2005 to January 2006. Three-dimensional swirl angle program control restitution bed was applied. This bed was composed of fixing chest swirl section,magnetic heat lumbar flexure massage section and traction haunch angulation section with the pull strength of 20-90 kg. ②The patients were divided into experimental group(n =38) and control group(n =33). ③All patients of the two groups received supine genuflex traction for 20 minutes and combined with longitudinal rotation for 10 minutes. Patients in the experimental group received 20? traction from below lumbar to bed,while patients in the control group received plane traction combined with longitudinal rotation. ④Lumbar function was determined by criterion of the lumbar disease made by Japanese Shaping Association to observe improved indexes and times of treatment. RESULTS:Totally 71 patients were involved in the result analysis. No significant difference was found in preoperative lumbar function between experimental group and control group(13.684?1.662,13.485?3.001,P

Objective To detect the protective mechanism of trehalose in tracheal cryopreservation.Methods Inbred male Sprague-Dawley (SD) rats were sacrificed with intraperitoneal injection of ketamine(150mg?kg -1).The tracheas were removed and immersed immediately in the freezing medium of low potassium dextran (LPD) solution only(Group Ⅰ) ,containing with 10% dimethylsulfoxide(DMSO)(Group Ⅱ), containing with 0.15mol?L -1 trehalose (Group Ⅲ),and containing with 10% DMSO and 0.15mol?L -1 trehalose (Group Ⅳ) respectively. A sterile plastic tube containing a 1-cm-long trachea was filled with the freezing medium,sealed,and frozen to -80℃ at rate of -1℃ per minute in a programmable freezer.Then the tube was stored in liquid nitrogen(-196℃) for 20 days. Then the specimen was thawed in a 37℃ water bath and rinsed with physiologic saline solution 10 times.Histologic changes before cryopreservation and after thawing were examined in each group. After the specimens were embedded in paraffin,5-(m-thick sections were stained with hematoxylin and eosin.The epithelium and cartilage was assessed. We also observed Bcl-2 and Bax gene expression by immunohistochemistry. At last, some tracheas(SD) after cryopreservation were thawed and transplanted into the abdominal cavity of Wistar rats. The transplanted tracheas were retrieved and assessed histologically.Results Microscopic findings of the tracheas in Group Ⅲ and Group Ⅳ showed their structure were intact and Bax gene expression was lower in cartilage after cryopreservation(20d) compared with other groups,especially in Group Ⅳ.The tracheas in Group Ⅲ and Group Ⅳ grew well after they were transplanted into cavity of Wistar rats heterotopically,too.There were no significant differences among 4 groups in Bcl-2 gene expression.Conclusion In tracheal cryopreservation the trehalose can protect the trachea by protecting the tracheal cartilage.It is one of the protective mechanism that the trehalose inhibit the Bax gene expression of cartilage cells.The concomitant use of trehalose and DMSO has a synergistic effect.

[Objective] To detect and genotype HPV in cutaneous wart tissue.[Methods] This study recruited 132 outpatients with cutaneous warts in the Affiliated Hospital of Zunyi Medical College.Of the 132outpatients,46 had verruca vulgaris,38 had verruca plantaris,and 48 had verruca plana.Ten healthy human and 15 patients with condyloma acuminatum served as the control.Tissue specimens were obtained from the lesions of these patients as well as from the normal skin of the healthy controls.DNA was extracted from the specimens followed by PCR amplification with general and type-specific primers.[Results] Of the 132 wart specimens,128 (97.0％) were positive for HPV,111 (84.1％) for HPV2,89 (67.4％) for multiple types of HPV,55 (41.7％) for two types of HPV,28 (21.2％) for three types of HPV,and 6 (4.6％) for four types of HPV.The most prevalent HPV types were HPV2 followed by HPV1 in verruca vulgaris and verruca plantaris,HPV2 followed by HPV10,HPV3 and HPVl in verruca plana.Multiple types of HPV were identified in 50.0％(23/46) of the verruca vulgaris specimens,76.3％ (29/38) of the verruca plantaris specimens,and 77.1％(37/48) of the verruea plana specimens.Among patients infected with more than one type of HPV,the combination of HPV1 and 2 was the most common (35,39.3％),followed the combination of HPV1,2 and 10(12,13.5％).[Conclusion]s The most prevalent HPV type appears to be HPV2,followed by HPV1,in patients with cutaneous warts in Zunyi area of Guizhou province.HPV2 infection is associated with various types of cutaneous warts.There is a high incidence of coinfection with multiple types of HPV in patients with cutaneous warts,especially in those with verruca plana.Moreover,HPV3 and 10 are newly identified in verruca vulgaris,HPV 10 in verruca plantaris,HPV 1 and HPV7 in veruca plana.

BACKGROUND:Traditionally, forced expiratory volume in one second (FEV1) was used to evaluate exercise response of patients with asthma; however, patients obviously had panting after exercise, so FEV1 was affected commonly. Impulse oscillometry (IOS) is a new technique for measuring respiratory impedance that do not require maximal inspiration and forced expiration.OBJECTIVE: To study airway resistance with IOS before and after exercise in healthy and asthmatic patients and investigate the significance of exercise excitation and IOS assessment.DESIGN: Synchronically non-randomized case contrast study.SETTING: Department of Respiratory Medicine, East Hospital Affiliated to Tongji University.PARTICIPANTS: A total of 14 male patients with bronchial asthma who were regarded as the asthmatic group were selected from Department of Respiratory Medicine of Shanghai East Hospital from January to October 2006. They were in a clinical stationary phase. Another 14 male healthy subjects were selected as the control group and ages of all subjects ranged from 29 to 50 years. All subjects provided the confirmed consent.METHODS: IOS was used to measure basic value of respiratory resistance, and then subjects underwent exercise challenge. Nose of subjects was clipped breathing through mouth. Within 3-4 minutes, heart rate was increased to 90% and maintained for 6 minutes during challenge. Respiratory resistance was repeatedly measured at 1, 5, 10, 15 and 20 minutes after exercise, including airway hyperresponse (AHR), total respiratory resistance, central resistance, peripheral resistance and resonance frequency at 5, 20 and 35 Hz of pulse frequency, elasrtic resistance and inertia resistance (X5 and X35) at 5 and 10 Hz of pulse frequency. In addition, difference of AHR at 5 and 35 Hz was calculated, and change ratios of both Rcentral and Rperipheral were calculated as (highest value after exercise-baseline value)/baseline value × 100%.MAIN OUTCOME MEASURES: Basic value of respiratory resistance by using IOS and exercise challenge test.RESULTS: All 14 patients with bronchial asthma and 14 healthy subjects were involved in the final analysis. Peripheral resistance (Rperiphera) was significantly higher than central resistance (Rcentral) in asthmatic patients (P ＜ 0.01). The maximal increase of respiratory impedance occurred from 5 minutes to 10 minutes after exercise in asthmatics. Resonance frequency (Fres) of asthmatics before and after exercise was significantly increased than that of controls (P ＜ 0.01).Change ratios of Fres from asthmatics were higher than that from control group (P ＜ 0.01). After challenge, R5, R5-R20,Zrespir and X5 from asthmatics changed significantly than that from controls (P ＜ 0.01). The increment change value of After exercise Zrespir increased significantly, because obstruction of small bronchi during expiration and impedance increased abruptly. Air trapping was expressed in VT-Zrespir graph in 57.1% patients. There was no difference in the VT-Zrespir graph of controls before and after exercise.CONCLUSION: The main site of airflow obstruction was in small airways in asthmatics after exercise challenge. The general acceptance of IOS method was good among the asthmatic patients. The airway response of exercise challenge may be assessed more accurately with IOS that do not require a maximal inspiration and forced expiration.

BACKGROUNDIn most instances, advanced non-small cell lung cancer (NSCLC) is treated with primary chemotherapy. Many chemotherapy regimens can palliate cancer-related symptoms. Quality of life and modestly improved survival are very important especially for elderly patients. This clinical trial is to compare the efficacy and toxicity of gemcitabine and oxaliplatin (GO) versus gemcitabine and cisplatin (GP) in treatment of advanced NSCLC in elderly patients.

METHODSA total of 42 patients with advanced NSCLC diagnosed pathologically were randomly divided into GO group (gemcitabine 1000mg/m² on days 1, 8; oxaliplatin 65mg/m² on days 1, 8) and GP group (gemcitabine 1000mg/m² on days 1, 8; cisplatin 30mg/m² on days 1-3), 28 days as a cycle. All patients received two cycles of chemotherapy at least.

RESULTSIn GO group, the response rate was 55.0%. Whereas in GP group, the response rate was 40.9%. The difference in response rate was not statistically significant between the two groups (P > 0.05). The median survival duration was 11.2 months in GO group and 11.8 months in GP group. The 1-year survival rate was 45% in GO group and 50% in GP group (P > 0.05). The main toxicities were well tolerated. Leukopenia and nausea/vomiting at grade III+IV, and alopecia and impaired renal function at grade I+II occurred more frequently in GP group than those in GO group (P < 0.05).

CONCLUSIONSBoth of the two regimens are feasible, well-tolerated and effective in treatment of advanced NSCLC in elderly patients. GO regimen may be safer than GP regimen.

Objective Retrospective study and report on cases of "symptomatic facet of residual bone mass" caused by percutaneous transforaminal endoscopic discectomy (PTED),to analysis of its causes and revision strategies.Methods Seven cases of "symptomatic facet of residual bone mass" after PTED were found in six medical centers from July 2015 to November 2017.Weintroduced the course of diagnosis and treatment,to analysis of the causes,clinical features and revision strategies of the rare complication.Results Seven patients came from different medical centers (2 cases in Ningbo No.6 Hospital and 1 case in each of the other medical centers).The average age of the subject is 67.29±9.64 years (range from 57-83 years).Among them there were 1 male and 6 female.PTED was performed for all cases with lumbar disc herniation or stenosis.The operative segments were 1 of L2,3,2 of L3,4,3 of L4,5,1 of L5S1.Symptoms occurred immediately after surgery in all cases except one after a week of operation and another one month later.Two cases were appeared symptom of contralateral irritation,and the rest were aggravated by the original symptoms.Two cerebrospinal fluid leakage caused by bone mass piercing the dural sac.The bone mass compressed the nerve root and caused 1 case of lower limb muscle weakness.Foraminoplasty was performed during PTED in all patients.After CT scan,5 cases of bone mass were found on the same side of operation,and 2 cases were in the contralateral side.The shortest time for revision was 2 days and the longest 3 months.After conservative treatment,the symptoms were relieved in only one case.Revision surgeries were performed for all the other 6 cases,2 with microendoscopic discectomy (MED),1 mobile microendoscopic discectomy (MMED),1 small incision operation,1 PTED and 1 with minimal invasive surgery of transforaminal lumbar intervertebral fusion (MIS-TLIF).The VAS scores of low back pain and leg pain was significantly relieved from 8.67±0.52 to 1.50±0.55.Conclusion FTED may lead to residual bone mass in lumbar foraminoplasty.The penetration of the bone mass block into the spinal canal can cause the compression symptoms of the corresponding segment.The patients showed the corresponding spinal canal stenosis and nerve root irritation symptoms.A revision operation is required to remove the oppressed bone mass to relieve the symptoms as soon as possible if the conservative treatment not effective.