Objective: To observe the clinical effect of puncturing Back-Shu points in treating chronic fatigue syndrome. Methods: Twenty-two subjects were recruited and treated by puncturing corresponding Back-Shu points based on syndrome differentiation. The short-form General Health Survey (MOSSF GHS) and the Chalder Questionnaires for Fatigue were adopted for evaluating the therapeutic effects. Results: Of the 22 patients, 4 cases showed a marked effect, 11 got effect, 7 got failure, and the total effective rate was 68.2%. Conclusion: Puncturing Back-Shu points is effective in relieving the symptoms of chronic fatigue syndrome and enhancing the patients' health standard.

Objective To discuss the value of video-assisted thoracoscopy plus minithoracotomy (VATM) in the management of thoracic diseases. Methods An 1.5 cm incision was made along the middle axillary line at the 7th costal interspace on the diseased side to introduce the thoracoscope. After the exploration of intrathoracic lesion, another incision 5~8 cm in length close to the lesion was made. The thoracic cavity was opened by way of the costal interspace. Surgical performance of exploration, dissection, hemostasis and suture was performed under thoracoscope and direct vision by using routine surgical instruments and thoracoscopic instruments. A total of 43 cases of VATM were carried out. Results The operation time was 40~150 min (mean, 67 min). The length of incision was 5~8 cm (mean, 6 cm). The chest drainage lasted 2~5 d. No postoperative complications occurred. The postoperative hospital stay was 5~8 d (mean, 6 d). Follow-up found no recurrence for 3~22 months in 18 cases of spontaneous pneumothorax and pleural effusion, and found no recurrence, distant metastasis or incision implantation for 5~20 months in 3 cases of lung cancer receiving either lobectomy or wedge resection. Simple biopsy of tumor was conducted in 6 cases of lung cancer, who obtained chemotherapy or gamma knife radiotherapy postoperatively and survived 5~21 months. Conclusions VATM is practical, minimally invasive and safe. By using routine surgical instruments it has an optimistic outlook.

Objective To explore the effect of different controlled hypotension method on the blood flow of the vertebral vein by measuring of blood flow of vertebral vein in rabbits. Methods Eighteen New Zealand rabbits,weighing 2-3 kg,were rando mly divided into three groups (n =6 each).Group S maintained MAP (90±5)mm Hg,group GTN reduced MAP to the base value of (70±10)% by using nitroglycerin 0.5 μg·kg-1 ·min-1 and esmolol 2.5 μg·kg-1 ·min-1 (ni-troglycerin∶esmolol= 1∶5 ),group SNP reduced MAP to the base value of (70 ± 10 )% by using sodium nitroprusside 0.5 μg·kg-1 ·min-1 and esmolol 2.5 μg·kg-1 ·min-1 (sodium nitroprusside∶es-molol=1∶5).The controlled hypotension model was established after intravenous general anesthesia. The blood flow of vertebral vein in rabbits were measured by ultrasonic measurement system (Terason 2 000 system ) before controlled hypotension and 1 hour after controlled hypotension. Results The MAP before controlled hypotension in group S (89.0 ± 5.2 )mm Hg,group GTN (91.5± 9.6 )mm Hg,group SNP (92.0 ± 5.7 )mm Hg had no significant difference.The blood pressure before and after the experiment had no significantly difference in group S.The blood pressure after controlled hypotension had no significant difference in group GTN and group SNP,but lower than that before controlled hypotension (P <0.05).Compared with group S,the blood flow of verte-bral vein in group GTN and group SNP were significantly reduced (P < 0.05 ).The blood flow in group GTN was significantly reduced compared with that in group SNP (P <0.05 ).Conclusion In the experiment,the combination of nitroglycerin and esmolol can better reduce blood flow of vertebral vein than that of nitroprusside and esmolol,that it is suitable for the control of hypotension in spinal surgery.

Objective To investigate the effects of health-education on the behaviors of following doctor's instructions of the patients receiving anticoagulation after valve replacement. MethodsThirty patients who received anticoagulation after valve replacement took part in this study. Health-education for knowledge of anticoagulation after valve replacement was implemented among these patients, and questionnaires investigating compliance for medication were subjected to all cases after their discharge of ward. The awareness of knowledge of anticoagulation was compared between patients of pre- and post-health-education,and the compliance of medication after discharge was also analyzed. ResultsHealth education improves the patient's understanding and knowledge about anticoagulation and also improves their attention to the doctor's instructions. Conclusion Health-education can increase the awareness of the knowledge of anticoagulation and the compliance among patients.

Objective To construct and express human ScFv against keratin in E.coli,and identify its binding activity with antigens and antigenic specificity.Methods By genetic engineering technology,human anti-keratin ScFv was constructed from Fab fragment selected from established semi-synthetic phage antibody library.The binding activity with antigens and antigenic specificity of expressed products were i-dentified by ELISA.Results Results of DNA sequence analysis showed that nucleotide sequence of V?and VH of ScFv gene was similar to that of V?and VH of Fab gene,suggesting that there was no mutation in the construction of ScFv.The soluble anti-keratin ScFv was found to have a good antigenic specificity as well as excellent binding activity with target antigens by ELISA.However,the binding activity of ScFv with keratin was lower than that of Fab.Conclusions The successful expression and identification of human an-ti-keratin ScFv might lay a solid foundation for further observation of biological activities of anti-keratin au-toantibody,manufacture of genetic-engineering anti-keratin products,and promotion of clinical application of genetic-engineering anti-keratin antibodies.

Objective To study the effects of anti-keratin autoantibody (AK auto Ab) on apoptosis of human keratinocytes. Methods Light and electron microscopy were used to observe morphological changes, flow cytometry (FCM) to analyze cell cycle, and electrophoresis to analyze DNA profiles of cultured keratinocytes. Results Typical morphological changes with apoptotic characteristics such as karyopyknosis, chromatin agglutination and apoptosis bodies were found. The subdiploid peaks due to apoptosis were also found in cell cycle analyses. DNA electrophoresis of keratinocytes showed characteristic ladder. Conclusions AK auto Ab could induce apoptosis in cultured human keratinocytes.

Objectives To investigate the binding of natural IgM to Staphylococcus aureus and its role in the phagocytosis of S. aureus by phagocytes, and to pave way for further study on the role and mechanism of natural IgM in defense of bacteria. Methods The binding of natural antikeratin IgM 3B4 to S. aureus was analyzed by ELISA and indirect immunoiluorescence. The role of 3B4 in the phagocytosis was analyzed by colony forming assay and flow cytometry (FCM). Results Both ELISA and indirect immunofluorescence proved the binding of natural IgM 3B4 to S. aureus. Colony forming assay found that the amount of colony forming units decreased significantly when 3B4 was added. The analysis of FCM showed that 3B4 augmented phagocytosis of 5. aureus by phagocytes. Conclusions Natural antikeratin IgM 3B4 can bind to S. aureus and regulate the phagocytosis of it, indicating that natural IgM may play some role in the defense against bacterial infection.

Objective To design and testify a novel strategy for acquiring mimetic epitope mapping by screening for a phage random peptide library using polyclonal anti keratin autoantibodies (AK auto Ab). Methods AK auto Ab were isolated and purified from pooled human sera by keratin affinity column in which keratin had been linked with CNBr Sepharose 4B,then biotinylated by the biotin ester. A 15 mer phage random peptide library was biopanned for 3 cycles and positive clones were identified by ELISA,competition assay and DNA sequencing. ResultsBy sequence comparison 23 positive clones were selected randomly and three epitopes were confirmed. Among the three epitopes SLSPMPTTNRR was the dominant epitope. The phages carrying positive clones reacted with AK auto Ab specifically and keratin could prevent interaction between AK auto Ab and positive phages. Conclusion The designed strategy is successfully applied in acquiring epitopes of polyclonal autoantibodies to keratin, which could provide a new approach for the discovery of epitope mapping which binds to natural autoantibodies.

Objective To immunologically characterize the binding activity of a genetic engineering human antibody against keratin. Methods The specific anti-keratin Fab was screened from a phage antibody library. After Fab monoclonal antibody was induced by isopropylthio-?-D-galactoside (IPTG), the binding activity with antigens and antigenic specificity of the antibody were verified by ELISA,Western blot, and competitive inhibition ELISA. Results The antibody possessed good antigenic specificity as well as excellent binding activities with antigens, and could recognize 46 000 keratin specifically. It was identified as anti-keratin 17 antibody. Conclusions The immunologic characteristics of the genetic engineering antibody are very good. It is necessary to further study the biological activities and clinical application of it.

Objective To prepare natural anti-keratin IgM monoclonal antibody. Methods Spleen cells of BALB/c mice raised in specific pathogen free conditions were directly fused with Sp2/0 myeloma cells. The hybridoma supernatants were tested by ELISA using pre-extracted keratin. The natural IgM obtained was further identified by immunochemistry and immunoblot methods. Results The cell fusion rate was about 60% without pre-immunization. About 14% supernatants reacted with the keratin antigen. Three hybridoma strains secreting natural IgM monoclonal antibody against keratin were obtained. The immunochemistry results showed that the natural anti-keratin IgM was able to bind to epidermis, sebaceous gland, hair follicule, and muscle tissues. Conclusion B lymphocytes in normal BALB/c mice spleen can produce natural antibody against kerain.