Objective To build an HPLC-ELSD method for simultaneous determination of 8-O-acetylharpagide and harpagide inAjuga decumbens Thunb.;To determineAjuga decumbens Thunb. samples from different area and of different batches.Methods Chromatographic separation was achieved on a Dikma C18 column (4.6 mm×150 mm, 5μm). The mobile phase consisted of water (A) and acetonitrile (B), using a gradient elution of 5% B at 0-6 min, 5%-15% B at 6-11 min, 15% B at 11-17 min, and the flow rate was 1.0 mL/min. An evaporative light scattering-detector (ELSD) was used with the temperature of drift tube at 103℃ and the gas flow rate of air was at 1.6 L/min.Results Good linearity was shown at the concentration range of 0.017-17.32μg for 8-O-acetylharpagide, and 0.003-1.62μg for harpagide. The validated method was of great precision and accuracy. The contents of 8-O-acetylharpagide and harpagide inAjuga decumbens Thunb. from Fujian province were higher than the contents in other localities.Conclusion The content of harpagide can be used as index component to monitor quality control ofAjuga decumbens Thunb..

Chinese compound priscriptions of traditional herbs are widely used in the alternative therapy for all kinds of diseases It remains unclear how the compounds in different herbs interact each other and which exacting mechanism can complain their effects The components and pharmacological effects of the combination of Chinese herbs are not due to the simple mixture of single herb's The substance basis research of combination herbs is one of the important issue for understanding their pharmacological mechanism Here the recent progress on the substance basis research of combination Chinese traditional herbs is reviewed, including the combinative effects and quantity ratio relation study, herbs serum study, chemical compounds research, the methods and techniques in chemical components analysis, and application foreground of Chinese compound priscription

Objective: : T o study the microanatomy of the facial nerve in the cerebellopontine angle (CPA) a nd the internal auditory meatus (IAM) for the preservation of the facial nerve i n the acoustic neurinomas surgery. Methods: Forty sides of CPA f acial nerve of 20 adult cadaver heads were examined. Results: In the pontomedullary sulcus the facial nerve (1.98? 0.10) mm anterior to the vestibulocochlear nerve,(8.76?1.42) mm lateral to the abducent nerve at the points where the nerves join the brain stem at the lateral end of the sulcu s, (8.15?2.18) mm above the junction of the Ⅸ nerve with the medulla. There were 62 nutritious artery supplying the facial nerve in the CPA, mainly from the anterior inferior cerebellaris artery (AICA) and its branches; and 17 in the IA M were mostly from the internal auditory arteria (IAA). Conclusion: To be familiar with the microanatomy of the facial nerve in CPA and IAM is h elpful for localizing the facial nerve and improve its preservation in acoustic neuroma surgery.

Objective To construct and identify recombinant vaccine Bifidobacterium bifidum (pGEX-Sj32) of Schistosomajaponicum (Sj).Methods The Sj32 gene amplified by PCR from template of plasmid pET28α-Sj32 that extracted from recombinant Escherichia coli(E.coli) BL21 (pET28α-Sj32) stored in our laboratory was cloned into E.coli-Bifidobacterium bifidum shuttle expression vector pGEX-1λT to construct recombinant plasmid pGEX-Sj32.The recombinant plasmid was transformed into E.coli BL21 (DE3).The plasmid was extracted and identified by restriction enzyme digestion.The recombinant Bifibacterium bifidum (pGEX-Sj32) vaccine was constructed by electroporating pGEX-Sj32 into Bb.The extracted plasmid was amplified and identified by PCR.Results The gene Sj32 of 1 270 bp in length was amplified by PCR.The restriction endonuclease digestion showed that the length of plasmid vector was 4 947 bp,and Sj32 gene was 1 270 bp.The Sj32 gene amplified by PCR from the template of pGEX-Sj32 extracted from recombinant Bb (pGEX-Sj32) vaccine was 1 270 bp in length which consistent with expected result.Conclusion The recombinant Bb (pGEX-Sj32) vaccine of Sj is successfully constructed.

Objective To investigate the clinical curative effect of closed thoracic drainage by minimal invasive with needle catheter in treating spontaneous pneumothorax.Methods Seventy-six patients with spontaneous pneumothorax were divided into two groups randomly,the control group was treated by traditional thick tube closed thoracic drainage,the experimental group was treated by the needle catheter minimally invasive thoracic closed drainage.Compared clinical curative effect,pulmonary reexpansion time,the wound healing time,the degree of pain and the occurrence of complication between two groups.Results The effective rate of experimental group and control group was 91.9％ (34/37) and 92.3％ (36/39),the pulmonary reexpansion time of experimental group and control group was (5.38 ± 2.21) d and (5.29 ± 2.27) d,there was no significant difference (P ＞ 0.05).The pain scores during draining and wound thealing time of experimental group and control group was (1.32 ±0.92) scores,(2.93 ± 1.14) scores,(2.13 ± 1.40) d and (5.32 ± 1.78) d,there was significant difference (P ＜ 0.01).Complication such as subcutaneous emphysema,dislodgment of the catheter and wound infection in experimental group was lower than that in control group.Conclusions The effect of needle catheter minimally invasive thoracic closed drainage in spontaneous pneumothorax is better,simple and less invasive.The wound healing time is short,and little pain.It is worthy of promotion.

Objective To construct the recombinant plasmid pGEX-Sj32 of Schistosoma japonicum(Sj )and to research its ex-pression in Escherichia coli (E.coli)BL21.Methods Sj32 gene was amplified by PCR from template of plasmid pET28α-Sj32 ex-tracted from recombinant bacterium BL21 (pET28α-Sj32 )stored by our laboratory,and then cloned into the vector pGEX-1λT to construct pGEX-Sj32.The recombinant plasmid pGEX-Sj32 was transformed into E.coli BL21(DE3).The recombinant strains were induced by isopropyl-β-d-thiogalactoside(IPTG),and the expressed products were identified by SDS-PAGE and Western blot.Re-sults Sj32 coding gene was successfully amplified by PCR and cloned into the vector pGEX-1λT,and the recombinant plasmid pGEX-Sj32 was constructed successfully.The molecular mass of the expressed recombinant protein was proximately 58 000 as de-tected by SDS-PAGE.The amount of the expressed protein was about 21% of the total bacterial protein.Western blot confirmed that the expressed protein could be recognized by the immune sera from rabbit infected with Schistosoma japonicum.Conclusion The recombinant plasmid pGEX-Sj32 is successfully constructed.The Sj32 protein was highly expressed in E.coli and the expressed recombinant protein possesses the specific antigenicity.

To observe the dynamic changes of splenocyte proliferation ,subsets and apoptosis in mice immunized with re-combinantBifidobacteriumbifidum(pGEX-Sj26GST)ofSchistosomajaponicum,themiceweresubcutaneously(SCgroup) and intranasally (IN group) immunized ,respectively .Four mice from each group were sacrificed in every 2 wk during 0-20 wk after immunization .Splenocyte proliferation was investigated by MTT colorimetric assay ,subsets of CD+4 and CD+8 T cells and apoptosis of splenocytes by FACsort flow cytometry .In SC group ,unstimulated and stimulated with S jAWA ,the level of splenocyte proliferation significantly increased at 4-20 wk after vaccination and increased markedly at 4-18 wk stimulated with ConA ,both of which peaked at 8 wk;in IN group ,the proliferation level of splenocyte cultured with SjAWA and ConA signif-icantly increased during the 4-18 wk ,2-10 wk and 14-18 wk ,2-8 wk and 12-18 wk ,respectively ,and all reached the maximum at the 4 wk after immunization (P＜0 .01 or P＜0 .05) .CD+4 subsets increased obviously during 2-14 wk ,2 wk and 6-16 wk re-spectively ,and reached the peak at 8 wk (P＜0 .01 or P＜0 .05) in both group ,while CD8+ subsets rose lightly during 2-20 wk in both group ,and reached the maximum at 8 wk (SC group) and 6 wk respectively (P>0 .05) .Whether unstimulated or stimulated with ConA ,the level of splenocyte apoptosis of which remarkably increased at 2-4 wk and 2-6 wk separately in SC group ,and both peaked at 2 wk (P＜0 .01 or P＜0 .05 );in IN group ,the level of splenocyte apoptosis all increased at 4 wk and reached the maximum at the same time .In summary ,by inducing the proliferation of splenocytes ,increasing CD4 + T cells and decreasing splenocyte apoptosis ,the rBb (pGEX-Sj26GST ) vaccine plays a critical role in the protective immune response .

Objective To construct and express a recombinant plasmid pGEX-Sj 14-3-3-Sj32 of Schistosoma japonicum in Escherichia coli (E.Coli) BL21 (DE3).Methods Sj14-3-3 and Sj32 antigen genes were amplified by PCR from template of plasmids pGEX-Sj14-3-3 and pET28α-Sj32 which were extracted from recombinant bacteria BL21 (pET28α-Sj32) and BL21 (pGEX-Sj14-3-3) stored in Institute of Infectious and Parasitic Disease of the First Affiliated Hospital of Chongqing Medical University.Sj14-3-3-Sj32 fusion gene obtained with gene SOEing was cloned into the vector pGEX-1λT to construct pGEX-Sj14-3-3-Sj32 which was identified by double digestion.The recombinant plasmid pGEX-Sj 14-3-3-Sj32 was transformed into E.Coli BL21 (DE3).The recombinant strains were induced by isopropyl-β-d-thiogalactoside (IPTG),and the expressed products were analyzed and identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting.Results The 1 750 bp Sj14-3-3-Sj32 fusion gene was successfully amplified by gene SOEing and cloned into the vector pGEX-1 λT verified by restriction analysis,the recombinant plasmid pGEX-Sj 14-3-3-Sj32 was successfully constructed.The molecular mass of the expressed recombinant protein was proximately 73 × 103 as detected by SDS-PAGE.Western blotting confirmed that the expressed protein could be recognized by the immune sera from rabbit infected with Schistosomajaponicum.Conclusion The recombinant plasmid pGEX-Sj14-3-3-Sj32 is successfully constructed and could be highly expressed in E.coli and the expressed recombinant protein has specific antigenicity.

Objective To introduce a new technique about reconstruction of anterior cruciate ligament(ACL) by transplantation of autogenous hamstring and fixation with a crossbar(Transfix Ⅱ) under arthroscopy and explore its effect on short term. Methods 72 patients with ACL injury were treated with ACL reconstruction by transplantation of autogenous semitendinous and gracilis tendon, fixed by a crossbar in femur end and a bio-absorbable interface screw in tibial tunnel. 2 cases of meniscus neoplasty, 31 partial meniscectomy, 8 large partial meniscectomy, 16 cartilage trimming, and 15 medial collateral ligament neoplasty, as well as 1 patellar tendon neoplasty were performed simultaneously. Pre- and post-operative knee joint function and stability as well as X-ray photograph were evaluated according to the Lysholm scoring scale system. The reliability of the fixation and its short-term effect were analyzed. Results 65 patients were available for follow-up with the duration from 8 to 36 months. The function of knee joint improved greatly. 8 cases had positive finding in physical examination. The average pre- and post-operative Lysholm score were 56?6.3 and 91?4.8, respectively(P