Evodiamine is one of the most important antitumor alkaloid from evodiamine. This study focused on the mechanism of evodiamine in inducing apoptosis of gastric cancer SGC-7901 cells through mammalian target of rapamycin (mTOR) signal pathway, in order to explore its antitumor mechanism and lay a foundation for clinical treatment of gastric cancer. The sole cytotoxic effect of evodiamine on SGC-7901 cells and human peripheral blood mononuclear cells (PBMCs) was observed by MTT assay. After the cells were respectively intervened with single evodiamine or evodiamine combined with z-VAD-fmk, the gene expressions of mTOR, p70S6K and 4EBP1 were analyzed by real-time PCR, and the protein expressions of mTOR and p-mTOR were detected by western blot. The result showed that evodiamine inhibited the apoptosis of SGC-7901 cells in a time-dependent manner, with no cytotoxic effect on human PBMCs. After the respective intervention with single evodiamine or evodiamine combined with z-VAD-fmk, the cells became round and floated in medium. Compared with the control group, both treatment methods can inhibit mTOR, 4E-BP1 and p70S6K gene expressions, with significant differences. Compared with single evodiamine, evodiamine combined with z-VAD-fmk showed a higher inhibitory rate in gene expression. According to the Western Blot result, evodiamine can inhibit the protein expressions of mTOR and p-mTOR regardless of the combination with z-VAD-fmk, with a higher inhibitory rate after z-VAD-fmk blocked caspase. In conclusion, evodiamine may promote the apoptosis of SGC-7901 cells through mTOR signal pathway.
Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Quinazolines ; pharmacology ; Signal Transduction ; drug effects ; Stomach Neoplasms ; drug therapy ; genetics ; metabolism ; physiopathology ; TOR Serine-Threonine Kinases ; genetics ; metabolism

OBJECTIVETo explore the methods and therapeutic effects of trans-fibular anterior-lateral approach combined with external fixation in the treatment of Gustilo III distal tibiofibula fractures.

METHODSFrom 2007 to 2010,9 patients including 7 males and 2 females with the mean age of 40 years(ranging from 29 to 51 years). All patients received internal fixation of fibula after debridement on the first phase, external fixator were used to fix tibia across ankle joint, and removed after successful skin graft; The second phase tibia was used to fix through the lateral incision used in phase I. Early functional exercise was encouraged ,the union condition and functional results of the ankle joint was evealuated. The criteria of the AOFAS Foot and Ankle Surgery was used to evaluate the effects.

RESULTSAll patients were followed up,and the duration ranged for 8 to 37 months(averaged 21 months). Nine patients were achieved bony union, the average healing time was 24 weeks. No plate rupture or screw loosening was found. According to the AOFAS Foot and Ankle Surgery evaluation system, 3 cases got excellent results, 4 good cases and 2 fair.

CONCLUSIONTrans-fibular anterior-lateral approach combined with external fixation for Gustilo III distal tibiofibula fractures can receive satisfactory reset, debond ankle joint eralier and imporove the clinical effects.

Adult ; Female ; Fibula ; Fracture Fixation ; methods ; Fractures, Bone ; diagnostic imaging ; surgery ; Humans ; Male ; Middle Aged ; Radiography ; Tibia ; Treatment Outcome

OBJECTIVETo investigate the effects of locking plate fixation through lateral deltoid approach for proximal humeral fracture combined with micro-invasive percutaneous plating (MIPPO) technique.

METHODSFrom April 2009 to March 2012,26 patients with proximal humeral fractures were treated with proximal humeral locking system plate fixation through lateral deltoid approach, including 17 males and 9 females with an average age of 58 years old ranging from 28 to 76 years old. The time from injury to operation was 3 to 10 days (averaged 5.6 days). According to Neer typing for the proximal humeral fractures, 7 cases had 2 parts of fracture,15 had 3 parts of fracture,and 4 had 4 parts of fracture. The Neer score for shoulder function was evaluated.

RESULTSAll patients were followed up,and the duration ranged from 10 to 21 months (averaged 13.6 months). All patients were achieved bony union,the average healing time was 12.5 weeks (ranged from 10 to 21 weeks). No humeral head necrosis and axillary nerve injury occurred. According to Neer scoring system,the total score was 88.36 +/- 7.82, pain 30.82 +/- 3.24, function 23.76 +/- 5.71, activity 17.59 +/- 5.36, anatomical position 7.03 +/- 2.39; the result was excellent in 18 cases, good in 5 cases, fair in 2 cases, poor in 1 case.

CONCLUSIONLateral deltoid approach combined with locking plate fixation for treatment of proximal humeral fracture has advantages of small invasion,less blood lossing, short operative time, stable fixation, high rate of fracture healing, and satisfactory functional recovery.

Adult ; Aged ; Bone Plates ; Female ; Fracture Fixation, Internal ; instrumentation ; methods ; Humans ; Humeral Fractures ; surgery ; Male ; Middle Aged ; Shoulder Fractures ; surgery ; Treatment Outcome

The gene of mutated TNF-?D4 gene was amplified by overlap PCR and cloned into the prokaryotic expressive vector pBV220.TNF-?D4 contains two changes:substitutions of Pro8Arg,Ser9Lys,Asp10Arg,Ile157Phe,Leu29Ser,Arg31Val and a deletion of the N terminal four amino acids.The recombinant vector pBV220-TNF-?D4 was transformated into E.coli strain DH5?,and the high expression strain was obtained by screening monoclones.The level of expression was about 45% of total cell protein.After purification,the purity of fusion protein was above 90% by HPLC and relative ability was 8 ?107.TNF-?D4 was modificated by mPEG-ButyrALD。After purification,the purity of mPEG-TNF-?D4 was above 85% and relative ability was 8.6?107.The in vivo systemic toxicity of mPEG-TNF-?D4,which is indicated by LD50,is lower than that of rhTNF-?.These results strongly supported for the further study and exploitation of TNF-antitumor drug.

The influenza A virus matrix protein2 gene(M2)which deleted transmembrane region was amplified by overlap extending PCR,and the multiepitope gene of hemagglutinin(HA)was PCR amplified with seven continuous synthesized segments by designing primer.The two gene segments were separately cloned into pMD18T vector to sequence analysis and prokarytic expression vector pET28a+ to construct the recombinant plasmid pET28a+M2dHA.The recombinant plasmid was transformed into E.coli BL21(DE3),and the high expression strain was obtained by screening monoclones.The recombinant protein existed as inclusion bodies,which accounted about 45% of the total cellular protein.The inclusion bodies were washed with 1% Triton X100 solution twice,and dissolved in 8 mol/L urea solution.The solution protein was purified by Ni+2 affinity chromatography,and refolded by dilution renaturation,then purified by Q Sepharose FF cation exchange column.The purity of the protein was over 90% by HPLC analysis.The result of Western blot showed it has good antigenicity and specificity.These results strongly supported for the further study of the broadspectrum influenza virus subunit vaccine.

Objective To compare the effect of cutaneous buckle and conventional suture on the suturing time and the artistic appear -ance of postoperative incision with patients who underwent anterior cervical spinal surgery .Methods The patients were randomly divided in-to cutaneous buckle group(50 patients) and conventional suture group(50 patients),the suturing time and artistic appearance of postopera-tive incision of two groups were observed .Results Buckle group and conventional suture group both got good wound healing ,the same heal-ing rate of two groups(100％) showed that there was no statistical difference in healing rate (P>0.05).The mean time of buckle group and conventional group were 7.3 minutes and 13.5 minutes,respectively,the suturing time of cutaneous buckle group was much shorter than con-ventional suture group (P<0.05).The satisfactory degree for artistic appearance of the former was higher than the latter (9.1 vs.6.8 points),the difference was significant(P<0.05).Conclusion Compared with conventional suture ,cutaneous buckle has the advantages of shorter suturing time ,better artistic appearance of incision and higher satisfactory degree in the anterior cervical spinal surgery .

Adolescent ; Adult ; Anastomosis, Surgical ; methods ; Female ; Finger Injuries ; surgery ; Humans ; Male ; Middle Aged ; Replantation ; methods ; Veins ; surgery

OBJECTIVETo study the clinical effect of arthroscopic anterior cruciate ligament (ACL) construction with different transplants.

METHODSFrom March 2006 to April 2009, 86 patients including 60 male and 26 female undergoing arthroscopic ACL reconstruction were prospectively randomized consecutively into autograft group (44 patients, using autogeneic hamstring tendons) and allograft group (42 patients, using allogenic lower extremity tendons). The age of those patients were 22 - 56 years, averaging (32 ± 7) years. The operations were made by the same doctor with the standard technology. The postoperative effects were assessed by the range of motion and tibia forward distance, Lachman test, pivot shift test, Daniel test, IKDC scores systems, Lysholm-Tegner scores.

RESULTSSeventy-nine patients were followed up, 41 patients in autograft groups averaged 39.6 months and 38 patients in allograft group averaged 37.4 months. The operation time of autograft group was (87 ± 11) minutes, that of allograft group was (55 ± 10) minutes (t = 15.732, P < 0.05). The time of postoperative fever of autograft group was (3.2 ± 1.4) days, that of allograft groups was (7.6 ± 5.3) days (t = 5.740, P < 0.05). The Lysholm scores of autograft group was 42 ± 7 before operation, and 89 ± 8 at final follow-up. The Lysholm scores of allograft group was 44 ± 6 before operation, and 87 ± 9 at final follow-up. There was statistic difference in both groups between before operation and final follow-up (t = 13.534 and 17.768, P < 0.05).But no statistic difference existed between the two groups (P > 0.05). The Tegner scores of autograft group was 2.9 ± 2.1 before operation, and 7.7 ± 1.2 at final follow-up. The Tegner scores of allograft group was 2.7 ± 1.4 before operation, and 7.1 ± 1.6 at final follow-up. There was statistic difference in both groups between before operation and final follow-up (t = 16.004 and 12.338, P < 0.05).No statistic difference existed between the two groups (P > 0.05). The KT2000 results showed that the anterior displacement of autograft groups was (10.7 ± 3.5) mm before operation and (5.0 ± 2.7) mm at final follow-up, the anterior displacement of allograft groups was (10.9 ± 2.9) mm before operation and (6.5 ± 2.4) mm at final follow-up, there was statistic difference between before and after operation in anterior displacement in two groups (t = 16.354 and 13.296 P < 0.05). There was no difference between two groups before operation and at final follow-up. Compared to before operation, the IKDC scores were improved greatly after operation (P < 0.05).

CONCLUSIONThe clinical effect of arthroscopic ACL construction with allograft transplants is near to autograft.

Adult ; Anterior Cruciate Ligament Reconstruction ; Arthroscopy ; Female ; Humans ; Knee Injuries ; surgery ; Male ; Middle Aged ; Prospective Studies ; Tendons ; transplantation ; Transplantation, Autologous ; Transplantation, Homologous ; Young Adult

OBJECTIVETo assess the clinical effect and safety of the Chinese medicine Longbishu Capsule combined with mesylate doxazosin in the treatment of benign prostatic hyperplasia (BPH) of the kidney deficiency and blood stagnation type.

METHODSThis was a randomized, double-blind, double-simulation control study. We equally assigned 60 men diagnosed with BPH of the kidney deficiency and blood stagnation type to an experimental and a control group, the former treated with mesylate doxazosin plus Longbishu Capsule and the latter with mesylate doxazosin plus placebo. We compared the International Prostate Symptom Score (IPSS), quality of life (QOL), Chinese symptom score (CSS), maximal urinary flow rate (Qmax), and prostate volume between the two groups of patients before and after 6 months of medication.

RESULTSAfter treatment, there were 5 cured cases, 13 markedly effective cases, 9 effective cases, 1 ineffective case, and 2 eliminated cases in the experimental group, as compared with 2 cured cases, 8 markedly effective cases, 10 effective cases, 7 ineffective cases, and 3 eliminated cases in the control group. The total effectiveness rate was obviously higher in the former (96.4%) than in the latter (74.1%). IPSS, Qmax, and CSS were improved in both of the groups after medication, even more significantly in the experimental than in the control group (IPSS: 15.22 ± 2.98 vs 18.15 ± 5.88, P <0.05; Qmax: [13.56 ± 2.26] ml/s vs [11.78 ± 2.97] ml/s, P <0.05; CSS: 6.18 ± 2.13 vs 9.52 ± 3.15, P <0.05). Because of the difference in the QOL score between the two groups at the baseline (P = 0.038 <0.05), no more comparison was made in this aspect after treatment.

CONCLUSIONThe combination of Longbishu Capsule with mesylate doxazosin is safe and effective for the treatment of BPH.

Adrenergic alpha-Antagonists ; therapeutic use ; Capsules ; Double-Blind Method ; Doxazosin ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Male ; Prostatic Hyperplasia ; drug therapy ; physiopathology ; Quality of Life ; Treatment Outcome ; Urination

OBJECTIVETo investigate the significance of Rac subfamily members in the gastrointestinal carcinogenesis and progression.

METHODSThe mRNA expression of Rac1, Rac2 and Rac3 in 12 kinds of gastrointestinal cancer cell lines was examined by semi-quantitative RT-PCR. The activities of Rac1 protein in 5 kinds of gastric cancer cell lines were tested by pull-down assay.

RESULTSCompared with the normal gastric mucosa and intestinal epithelial cell line, the mRNA expression of Rac1 and Rac3 was up-regulated in most of gastrointestinal cancer cell lines. The activities of Rac1 protein increased markedly in gastric cancer cell lines.

CONCLUSIONThe increased mRNA expression of Rac1 and Rac3 in gastrointestinal cancer cell lines and the abnormal activation of Rac1 protein in gastric cancer cell lines might be correlated with the carcinogenesis of gastrointestinal cancer.

Cell Line, Tumor ; Gastrointestinal Neoplasms ; metabolism ; Humans ; RNA, Messenger ; analysis ; Reverse Transcriptase Polymerase Chain Reaction ; rac GTP-Binding Proteins ; genetics ; rac1 GTP-Binding Protein ; analysis ; genetics