Objective To investigate the pharrnacodynamics of compound Chuanxiong granules in treating dysmenorrhea.Methods The effects of compound Chuanxiong granules on uterus contraction,anti-inflammatory,analgesia and hemostasis were researched by rat's uterine smooth muscle contraction test in vivo,the level of prostaglandins F2α effect test,mice torsion model induced by oxytocin test,mice auricle swelling model induced by dimethylbenzene test,and the coagulation time of mice test.Results Compared wAth the model control group,Chuanxiong granules may obviously inhibit smooth muscle constriction resulted from oxytocin (the average inhibition ratio in high-dose group,middle-dose group and low-dose group were 32.3％,27.2％,17.6％),decline the levels ofprostaglandins F2 [the value in high-dose group,middle-does group and low-dose group was (79.2±4.3)rig/L,(79.3 ±12.9) ng/L and (88.2±7.6) ng/L] inhibit twist times of mice (the value in high-dose group,middle-does group and low-dose group was 39.4％,46.8％,24.8％),inhibit the auricular swelling of mice caused by dimethylbenzene [the value in high-dose group,middle-does group and low-dose group were(31.7±21.0)％,(35.6±33.7)％,(59.3±35.0)％],and shorten the coagulation time [thevalue in high-dose group,middle-does group and low-dose group was(41.1±9.7)s,(43.3±10.1)s,(50.5±10.8)s respectively].Conclusion Compound Chuanxiong granules is a effective agent to relieve dysmenorrhea.

Objective: To fuse Mycobacterium tuberculosis protective antigen gene with mice ubiquitin gene, constructing antigen ubiquitin system. Methods: Mice ubiquitin cDNA was amplified by RT PCR from mice testicle,and 4 antigen genes were obtained by PCR from cultured Mycobacterium tuberculosis . Ubiquitin and 4 antigen genes were linked by flexible adaptor respectively and the fusing genes were cloned into pVAX vector.The recombinant plasmids were digested with endonuclease and sequenced.Then the recombinant plasmids were transfected into COS7 cells and the expression was assayed by ELISA. Results:Ubiquitin and 4 antigen genes were 0.2,0.3,0.7,1.0,1.65 kb in length by agarose electrophoresis. Endonuclease digestion of the recombinant plasmids indicated that the fusion genes were correctly inserted into pVAX vector. Sequencing results of fusion nucleic acid vaccines were identical to those in GenBank.The recombinant plasmids expressed in COS7 cells. Conclusion: Four Chinese Mycobacterium tuberculosis protective antigens ubiquitin systems are successfully constructed and can be expressed in eukaryotic cells. This may provide a basis for diagnosis and therapy of tuberculosis.

Objective: To clone porcine IL-4 cDNA and observe its adjuvanticity of vaccine against T. solium cysticerco-sis. Methods:The cDNA of porcine IL-4 was amplified by RT-PCR, which had 5' AUG initiatory codon with optimized trans-lational initiation. After sequencing,the cDNA was intergrated into expression vector pcDNA and transient expression was performed. Then newborn pigs were immunized with IL-4 expression vector and protective antigen DNA vaccine against T. solium cysticercosis. Results:The obtained sequence of porcine IL-4 cDNA was the same as reported. IL-4 and protective antigen could effectively promote the humoral response. Conclusion:An efficient expression plasmid containing porcine IL-4 cDNA is constructed. Its adjuvanticity of DNA vaccine against T. solium cysticercosis is preliminarily conformed.

Objective To analyse the immunostimulatory activity of CpG sequences in cysticercus cellulosae paramyosin (also named Antigen B,AgB)cDNA. Methods C57BL/6 mice were immunized with pcDNA3 AgB plasmid,pcDNA3 AgB′(CpG sequences were mutated),pcDNA3 or AgB protein and two weeks later,immune response was assayed by ELISA. Results IgG and IgG 2a were detectable at week 2 after immunization and continually increased until week 4.The antibody levels elicited by pcDNA3 AgB were significantly higher( P

Objective To clone and express the gene encoding ESAT antigen of Mycobacterium tuberculosis (chinese stain). Methods Genome of mycobacterium tuberculosis was extracted. Then, the full length cDNA encoding ESAT protein was amplified by PCR and cloned into prokaryotic expressing vector pGEX5T and sequenced. pGEX5T ESAT was expressed in k802 Ecoli, and the expressed protein was determined by western blot using the sera from ten patients with tuberculosis. Results One specific band of 0.3kb or so was obtained and sequencing result was identical to that reported from Genbank. The expressed protein could be specifically recognized by the sera from tuberculosis patients. Conclusions The full length cDNA of Mycobacterium tuberculosis (Chinese strain) ESAT protein was cloned and expressed successfully, which will be helpful in further studies on diagnosis and treatment of tuberculosis.

BACKGROUND:At present, there are many researches about repairing articular cartilage defects. In particular, the microfracture technique has been widely used. OBJECTIVE:To observe recovery of knee joint motor function and morphological changes in tissue repair during articular cartilage defects with different directions (coronal position and sagittal position). METHODS:Articular cartilage fracture models with 2 mm-thick medial femoral condyles of rabbit knee joint were established. According to incision directions, models were assigned to coronal and sagittal groups. At 5, 10 and 20 weeks after model induction, general observation was performed. Specimens were sliced into paraffin sections, and subjected to hematoxylin-eosin staining and col agen staining. Tissue repair at the articular cartilage defects was observed using optical microscope and immunohistochemical method. After model induction, range of motion of rabbit joints was regularly examined in the two groups.RESULTS AND CONCLUSION:A white line was seen across the femoral condyles at defects in the two groups. Articular surface at defect repair was at the level of in situ cartilage, and reached a bone union. Knee joint treated by operation did not affect function. Under light microscope, partial reconstruction of subchondral bone was seen in the two groups, mainly fibrocartilage repair. The level of bony remodeling was lower than tidal line of adjacent in situ cartilage. Immunohistochemical method exhibited that type I col agen staining gradual y reduced at defects of specimens, but type II col agen staining gradual y increased. These results suggested that there was no significant difference in the recovery of motor function of knee joint and the repair of articular cartilage with different directions (coronal and sagittal position).

Vinflunine tartrate-loaded liposomes (VT-L) with two drug-to-lipid ratios were prepared by pH gradient method. Vesicle size and zeta potential were determined by the Zetasizer Nano ZS. Entrapment efficiency was evaluated by cation exchange resin centrifugalization method. The toxicity and tumor inhibition to nude mouse administrated by VT-L with different drug-to-lipid ratios were investigated and compared with the vinflunine tartrate injection (VT-I). The results showed that the mean particle size, zeta potential and entrapment efficiency of the VT-L with drug-to-lipid ratios of 1 : 5 and 1 : 10 were 124.6 nm and 128.3 nm, -25.3 mV and -22.8 mV, 94.46% and 97.31%, respectively. The VT-L with two different drug-to-lipid ratios has significantly higher anti-tumor effect to nude mouse transplanted human non-small cell lung carcinoma A549 and lower toxicity than VT-I. While there were no significant differences in anti-tumor effect and toxicity between VT-L with two different drug-to-lipid ratios.

Objective:To compare the three effects of lidocaine in the prevention of general anesthesia in elderly patients.Methods:A total of 120 elderly patients (65-85 years old) underwent anesthesia with general anesthesia (ASA) Ⅰ ～ Ⅱ were randomly divided into thyrocricocentesis group (group H),throat surface anesthesia group (Group Y),intravenous injection group (group J) and control group (group D).Group H was injected with lidocaine for surface anesthesia;group Y used laryngeal spray for laryngeal sprayed lidocaine for surface anesthesia;group J was anesthetized induction of intravenous lidocaine to prevent intubation reaction;group D the control group was not treated with lidocaine.(SBP) and heart rate (HR) were measured before and after induction (T0),tracheal intubation (T1) and 1 (T2),3 (T3) and 5 min (T4),the changes of hemodynamics related indexes were compared.Results:Compared with the same group of T0,the SBP and HR of four groups of T1 moments were significantly decreased (P ＜0.05);In the other three groups,SBP was significantly increased at T2 and T3 (P ＜0.05),HR ofT2 was significantly increased (P ＜0.05);Compared with group D,SBP and HR in group H,Y and J were significantly different at T2,T3 and T4,he difference was statistically significant (P ＜0.05);but there was no statistically significant difference between the three groups (P＞ 0.05).Conclusion:The three ways of lidocaine can be used to prevent the general anesthesia reaction in elderly patients,the effect of three ways is parallel.However,cricothyroid membrane puncture increased the patient's pain,throat spray method increased the cumbersome operation and enhanced the cost of the patient,and intravenous injection method is simple and worthy of popularization and application in clinic.

BACKGROUND:Studies have shown that umbilical cord blood stem cel transplantation promote the recovery of spinal cord injury, and electroacupuncture also can inhibit the proliferation of astrocytes to reduce damage to scar formation, suggesting that a combination of umbilical cord blood stem cel transplantation and electroacupuncture may play an important role in the treatment of acute spinal cord injuries. OBJECTIVE:To observe the influence of transplantation of human umbilical cord blood stem cels combined with electroacupuncture at theDu channel on expression of nerve growth factor and neurotrophin 3 in rats with spinal cord injuries. METHODS: Seventy-two female Sprague-Dawlay rats were randomly divided into control group, injury group, transplantation group and combined therapy group. In the control group, only an incision on the back was sutured;in the injury group, a piece of saline-infiltrated gelatin sponge, 1 mm×2 mm×2 mm, was placed into the transected spinal cord at T10 level; in the transplantation group and combined therapy group, a piece of gelatin sponged infiltrated in the suspension of human umbilical cord blood stem cels was placed into the transected spinal cord, respectively, and then, electroacupuncture stimulation at the Duchannel was performed in the combined therapy group at 1 hour after modeling. Specimens were taken at 7, 14, 28 days after modeling in each group, and then immunohistochemistry, western blot and real time-PCR methods were used to detect the expression of nerve growth factor and neurotrophin 3. RESULTS AND CONCLUSION:Compared with the transplantation group, the expression of nerve growth factor and neurotrophin 3 was lower in the injury group but higher in the combined therapy group at 7, 14, 28 days after modeling (P < 0.05). The results of western blot and real time-PCR were consistent with those of immunohistochemical detection. Findings show that human umbilical cord blood stem cel transplantation combined with electroacupuncture has a remarkable synergistic effect in the treatment of spinal cord injury that can significantly up-regulate the expression of nerve growth factor and neurotrophin 3, and contribute to injured spinal cord repair, regeneration and functional recovery after spinal cord injury.

ObjectiveTo construct the eukaryotic coexpression plasmid CEA/IL-2, and to lay the foundation for further studying CEA nucleiotide vaccine , adjuvant and their effects of special antitumor immunity.MethodsThe eukaryotic expression plasmid (pcDNA3-CEA)containing the gene coding for CEA was obtained by RT-PCR and gene recombination techniques.Using enzymolysis,ligation and other techniques,an eukaryotic coexpression plasmid (pIRES-CEA/IL-2)containing two expression unites of CEA and IL-2 gene connected with internal ribosome site was constructed.ResultsThe coexpression plasmids were transformed into COS7 cells and expression of two proteins were demonstrated by ELISA, and flow cytometer and elecsy.CEA and IL-2 were (23.73?0.26)ng/ml,and(20.17?0.13)ng/ml respectively.ConclusionsThe eukaryotic expression plasmids pIRES-CEA/IL-2 could be successfully constructed and transformed into COS7 cells.Expression of two proteins were demonstrated with no difference on expression.