Objective To explore the effect of autologous blood transfusion on cytoimmunity during orthopedic operation.Methods Two hundred and twenty-six patients undertaken selective operation from June 2007 to October 2011 were divided into observation group ( 154 cases) and control group (72 cases).The observation group was received autologous blood transfusion and the control group was given homologous transfusion.T lymphocyte subsets and natural killer (NK) cell counts in blood samples were detected by flow cytometry before operation and 1,6 days after operation.Results CD3+,CD4+,CD4+/CD8+,NK cell of the observation group and control group on 1st day of postoperation were 0.7184 ±0.0921,0.3878 ±0.0611,1.64 ± 0.27,0.1627 ± 0.0633 and 0.6548 ± 0.0852,0.3137 ± 0.0726,1.18 ± 0.31,0.1465 ± 0.0514,respectively,which decreased obviously compared with those of preoperation in both groups (0.7436 ± 0.1069,0.4301 ±0.0818,1.68 ±0.31,0.1945 ±0.0572 and 0.7537 ±0.0940,0.4453 ±0.0608,1.62 ± 0.32,0.1821 ± 0.0571 ) (P ＜ 0.05 ).The above parameters of the observation group were significandy higher than those of the control group on 1 st day of postoperation (P ＜ 0.05 ).There were no significant differences of CD3+,CD4+,CD4+/CD8+ and NK cell on the 6th day of postoperation compared with the preoperation in the observation group (P ＞ 0.05).But CD3+,CD4+,CD4+/CDs+ and NK cell on the 6th day of the postoperation were 0.6266 ± 0.0905,0.3048 ± 0.0425,1.07 ± 0.27,0.1408 ± 0.0716,which were significantly lower than the preoperation in the control group (P ＜ 0.05).Conclusions Intraoperative autologous blood transfusion have less inhibitory effect on cytoimmunity than homologous transfusion.The patients having autologous blood transfusion have a rapid recovery of cytoimmunity.

Purpose: In order to observe the therapeutic effect of electric acupuncture plus Tuina therapy for cervicogenic headache. Methods: 200 cases of the patients with cervicogenic headache were allocated randomly into electric acupuncture plus Tuina group (Group A, n=100), electric acupuncture group (Group B, n = 50) and Tuina group (Group C, n = 50) and treated respectively by electric acupuncture on cervical Jiaji (Ex-B 2)points, Fengchi (GB 20),Ashi points, Shuaigu (GB 8), Yifeng (TE 17), Touwei (ST 8) and Waiguan (TE 5) on the sick side and Tuina therapy. Results: The total effective rate was 100% in Group A, 90.0% in Group B and 88.0% in Group C, with a significant difference (P＜ 0.05). Conclusion: Electric acupuncture plus Tuina therapy is an effective method for cervicogenic headache.

Objective To establish a new method for detecting pemphigus antibody (PAb). Methods PI 1 anti idiotype monoclonal antibodies against pemphigus and purified PAb from the serum of a patient with active pemphigus were used to establish an ELISA system for detecting the PAb. Results The result showed the purified PAb was IgG4 subclass, the sensitivity and specificity for the detection of PAb were high in the ELISA system, the sensitivity and specificity were not significantly different among the IIF, indirect ELISA and ABC ELISA. The standard curve for detecting the concentration of the purified PAb was primarily obtained in the study. Conclusion The ELISA system for the detection of PAb in the sera of patients is a good qualitative method, it might be of value in the clinical diagnosis of pemphigus. It is expected to quantitatively detect PAb of pemphigus patients in the future with the ELISA system established, which is directed to the IgG4 subclass of PAb, so it may be of value in the study of IgG4 subclass in the pathogenesis of pemphigus.

OBJECTIVE Learned through the seeing about the infection management of the Japanese Oita Medical University affiliated hospital and analyzed our hospital infection management′s status,to understand the dynamic management of hospital infection.Moreover these suggested that we attach importance to the management of hospital infection and constantly explore new ways of hospital infection management.METHODS According to the comparative presentation of these two hospitals′ awareness and practices of infection management from China and Japan,this paper probed into the direction of our hospital infection management development.RESULTS Hospitals in every country of the world have attached great importance to their infection management,but their specific practices were different.CONCLUSIONS By comparing inspection,every hospital should maintain the individualized characteristics and learn from each others′ strong points to offset their own weakness in order to gain continuous improvement of hospital infection management.

Objective To study the effect of human ?-globin matrix attachment region(MAR) on transgene expression in stably transfected CHO cells.Methods Expression vector was constructed,which contained the ?-globin MAR in both sides of Chloramphenicol acetyltransferase(CAT) reporter gene expression cassette in cis,then transfected into CHO cells.The CAT reporter gene expression was analyzed by ELISA method.Results The ?-globin MAR enhanced the CAT gene expression 5.5-fold in stably transformed CHO cells,while the transgene expression variation among individuals of transformants was decreased.Conclusion MAR increase transgene expression in stably transfected CHO cells.

Objective To screen and analyze genes differentially expressed within dermal papillae cells(DPC)with aggregative behavior.Methods Total RNA was extracted from DPC with and without ag-gregative behavior,and double-stranded cDNA were synthesized by using SMART cDNA synthesis.The cD-NA fragments of differentially expressed genes in dermal papillae cells with aggregative behavior were isolat-ed by suppression subtractive hybridization,sequencing,and then subtracted library was set up.Positive clones were screened by PCR method and verified by cDNA dot blot and then analyzed through homologous retrieving.Results A subtractive cDNA library of DPC with aggregative behavior was successfully construct-ed.The results of screening and cloning of the library showed that DPC with aggregative behavior could ex-press genes related to homologous aggregation,regnlation of growth,differentiation and development,and sig-nal transduction proliferation and cycle control,which included known genes(capping protein,paladin,vas-cular endothelial growth factor),hematopoietic stem/progenitor cells(HSPC)related genes(HSPC011and HSPC016)and a new gene.Conclusions The construction of subtractive library of DPC lays solid founda-tion for screening and cloning new and specific genes related to aggregative behavior of DPC.Several genes may cooperatively involve in homologous aggregation,and regnlation of growth of DPC.Among these genes,capping protein and palladin may be closely related to aggregative behavior of DPC,and VEGF and HSPC re-lated clones may be responsible for the status of higher proliferation of DPC.

BACKGROUND: Epidermal stem cells (ESCs) are of importance in the wound repair. Explaining the mechanism thatChinese herb speeds up the regeneration of injured skin from the angle of inducing stem cells deserves to be studied. OBJECTIVE: To observe the effect of Yuhong ointment on the proliferation and differentiation of ESCs during ratwound healing. DESIGN: A randomized controlled animal experiment. SETTING: Department of Pharmacology, Wangjing Hospital, China Academy of Chinese Medical Sciences. MATERIALS: This study was carried out in the Department of Pharmacology, Wangjing Hospital, China Academy ofChinese Medical Sciences between July and September 2006. Totally 114 healthy Wistar male adult rats, of cleangrade, weighing 180-210 g, were provided by the Laboratory Animal Center, Institute for Basic Theory of TraditionalChinese Medicine, China Academy of Chinese Medical Sciences were enrolled in this study. The processing ofanimals corresponded to the standard of Animal Ethics. Yuhong ointment was purchased from the PharmaceuticalCenter, Guanganmen Hospital, China Academy of Chinese Medical Sciences (Lot No. 20020110). Jingwanhong wasthe product of Tianjin Darentang Da'er Pharmcaceutical Co., Ltd (Lot No. Z12020440). METHODS: The 114 rats were completely randomly chosen and divided into 3 groups with 36 in each: Yuhongointment group, Jingwanhong group and model group, and the left 6 rats were involved as normal control group. Ratsin the normal control group were raised routinely, and no intervention was carried out. Immediately after beingmodeled, rats in the Yuhong ointment group were spread with Yuhong ointment, 0.1 g each wound, rats in theJingwanhong group were spread with Jingwanhong, 0.1 g each wound. Dressing change was daily carried out twice regularly until wound healing; The wounds of rats in the model group were untouched. MAIN OUTCOME MEASURES: On the 1st, 3rd, 5th, 7th, 14th and 21st days after modeling, wound healing time of rats in eachgroup was recorded, and integrinβ1 absorbance and transcription factor p63 positive cell amount were compared. RESULTS: Totally 114 rats were involved in the final analysis.① Wound healing time of rats in the Yuhong ointmentgroup and Jingwanhong group was significantly shorter than that in the model group, respectively (P ＜ 0.05). ② Integrinβ1 absorbance and transcription factor p63 positive cell amount of modeled rats in the Yuhong ointmentgroup, Jingwanhong group and model group were significantly higher than those in the normal control group, respectively (P ＜ 0.05); The above-mentioned two indexes in the Yuhong ointment group and Jingwanhong groupreached the peak on the 7th day, which was earlier than peak time in the model group, and peaked intensity of twoindexes was significantly higher than that in the model group, separately (P ＜ 0.05). CONCLUSION: Yuhong ointment can promote rat wound healing, which may be associated with Yuhong ointmentinducing the proliferation and differentiation of ESCS left in the wound edge.

Objective:To establish an HPLC method for the determination of patulin in health foods containing hawthorn to im-prove the quality control of patulin in related health foods. Methods:An Agilent TC-C18(2)(250 mm ×4.6 mm, 5 μm) column was used with the temperature of 30℃. The mobile phase consisted of 0. 8% tetrahydrofuran and the flow rate was 1. 0 ml·min-1 . The detection wavelength was set at 276nm. Results:The calibration curve of patulin was linear within the range of 1-20ng(r=1. 000 0), the average recovery was 92. 1% and RSD was 2. 2%(n=6). Conclusion:The method is simple, reliable and accurate, which can be used in the content determination of patulin in health foods containing hawthorn.

Objective To investigate the immortalization of human melanocytes by transfection with SV40 T antigen ( SV40T ). Methods By using SofastTM,a gene transfection reagent, the reconstructed eukaryotic expression vector SV40T-pEGFP was stably transfected into cultured primary human melanocytes, then the positive cells were selected with G418. After the positive cells were expanded in culture, the expression of SV40T gene was detected by RT-PCR and PCR, and the protein expression of SV40T by Western blotting. Results The genome DNA and total RNA were isolated from the positive cell clones, and a 288 bp fragment, which was specific for the SV40T antigen gene, was amplified. The results of immunohis-tochemistry and Western blotting confirmed the expression of SV40T protein in transfected cells. Conclusion SV40T antigen gene can successfully induce the immortalization of human melanocytes.

Objective To evaluate the feasibility and clinical efficacy of intra-abdominal exposure instruments in laparoendoscopic single-port nephrectomy(LESS-N).Method From February 2012 to July 2016,61 cases of LESS-N were performed in our center.There were 34 males and 27 females with a mean age of (60.3 ± 9.4) years old (ranging 36-72 years old).There were thirty-nine cases of renal tumors and twenty two cases of nonfunctioning kidney.The patients were divided into two groups.Group A included 39 cases that underwent conventional LESS-N (22 radical nephrectomy/17 simple nephrectomy).Group B included 22 cases that underwent intra-abdominal exposure instruments assisted LESS-N (17 radical nephrectomy/5 simple nephrectomy).The perioperative and postoperative data were collected and analyzed retrospectively.Results All the procedures of these two groups were completed successfully.In Group A,four patients were added one 5 cm additional trocar and two patients were converted to open surgery.No additional trocars or conversion to open surgery were needed in Group B.For LESS radical nephrectomy,there were no significant differences of mean tumor diameter (5.7cm vs.5.4 cm,P =0.65) between two groups.The average operative time was (95.1 ± 43.9) min in Group B which was lower than that in Group A (127.4 ± 61.9) min (P ＜ 0.01).The mean renal vascular processing time was declined from (25.4 ± 10.1)rmin in Group A to (18.8 ± 8.9)min in Group B (P ＜ 0.05).The mean estimated blood loss was (128.6 ± 51.1) ml in Group A and (98.7 ±-57.6) ml in Group B (P ＜ 0.05).No severe intraoperative and postoperative complications occurred in both group.Conclusions Intra-abdominal exposure instruments are feasible and effective for LESS-N.This system may shorten the operation time,reduce the amount of bleeding and improve surgical accuracy.