AIM: To explore the effect of Danshen on warfarin via detecting the patients with atrial fibrillation. METHODS: With the help of detecting prothrombin time(PT) and international normalized ratio(INR) of the patients who took warfarin only or both Danshen and warfarin,we compared the PT and INR of each group. RESULTS: At steady-state levels of warfarin,Danshen prolonged the PT(P

Objective To construct eukaryotic expression plasmid pEE14.1-dsFv?pr+,and detect the expression of the recombined gene in eukaryotic CHO-K1 cells.Methods The cationic DNA fragment was cloned into the 3' of VH gene by overlapping extension PCR,and the 6?His tab was inserted to the 3' of VL and human IFN-? gene by the same way.The above mentioned recombinant VH and VL genes were inserted into a pCI-GPI vector first,and then cloned into the pEE14.1 vector to construct the recombinant plasmid pEE14.1-dsFv?pr+.Finally,the recombinant plasmid was transfected into the CHO-K1 cells by LipofectamineTM 2000,and the expression was detected by RT-PCR,ELISA and Western blotting.Results The enzyme digestion and sequencing analysis showed that the recombinant plasmid was successfully constructed.RT-PCR showed that only the cells with transfected plasmid can generate the specific 1700bp fragment.ELISA analysis showed that the production of IFN-?expressed in the supernatant of transfected cells was about 1.1ng/ml.Also,Western blotting could reveal the characteristic band of HBsAg dsFv?pr+ protein.Conclusion The antibody targeting to human IFN-?genes has been successfully expressed in a single open reading frame.Changing the electricity of the antibody may provide the necessary condition for the study of the a new type of anti-HBV drug in nanoscale in the future.

Objective To construct the prokaryotic expression plasmid of human prostate stem cell antigen (PSCA),to induce the expression of GST-PSCA fusion protein in E. coli BL21,and to identify the purified recombinant fusion protein. Methods The fragment of PSCA gene was amplified by PCR,and then cloned into the pGEM-T Easy vector. The transitional plasmid Teasy-PSCA was identified by DNA sequencing. The PSCA gene was digested from the plasmid Teasy-PSCA by restrictive enzyme BamH I and Sal I,and then inserted into the pET42a vector which contains a glutathione s-transterase (GST) tag. Following the double restriction enzyme digestion,the recombinant plasmid pET42a-PSCA was obtained and transformed into E. coli BL21 (DE3). The expression of GST-PSCA fusion protein was induced with IPTG. The recombinant fusion protein was purified by passing over a Glutathione Sepharose 4B column,and was identified by SDS-PAGE and Western blotting. Results The length of amplified PSCA gene fragment was consistent with that expected,and the sequence was correct as exemplified by the PSCA gene reported in GenBank. The result of enzyme digestion indicated that the prokaryotic expression plasmid pET42a-PSCA was successfully constructed. After transformation with pET42a-PSCA and induction with IPTG,the recombinant target protein of about 43kD was obtained. The GST-PSCA fusion protein was correctly identified by SDS-PAGE and Western blotting. Conclusions The prokaryotic expression plasmid of human PSCA gene has been successfully constructed. The GST-PSCA fusion protein may express and be purified in E. coli BL21,and it lays a foundation for further study on the anti-prostate cancer gene vaccine.

BACKGROUND:The biomechanical properties of naturaly regenerated damaged articular cartilage that belongs to the fibrovascular tissue are far worse than those of the normal cartilage so that they cannot meet the requirements for joint function, leading to traumatic arthritis and loss of joint function.
OBJECTIVE:To evaluate the effects of matrix metaloproteinase-3 (MMP-3) inhibitor I with different concentrations on the early-stage repair of ful-thickness articular cartilage defects in the knee of rats.
METHODS: Twenty-four Sprague Dawley rats were randomized into control, defect (DEF), and defect combined with low-(D+L) and high-dose inhibitor (D+H) groups (n=6 for each group), respectively. Full-thickness articular cartilage defects followed by intraarticular injection of low- and high-dose MMP-3 inhibitor I for 4 weeks was administered in the later two groups. Serum MMP-3 was detected using ELISA method before and after experiment, respectively. Femoral trochleas were collected to observe characteristics of repaired tissue by gross appearance scoring and O’Driscoll histological scoring with Safranine O-Fast Green staining, and to measure type II colagen by immunohistochemistry after experiment.
RESULTS AND CONCLUSION:Rats in the D+H group had obvious repair similarly to hyaline articular cartilage, while creamy white cartilage tissue and fibrous tissue repair were observed in D+L group and in DEF group. D+H group obtained the best repair results according to gross appearance scoring and O’Driscol histological scoring and the highest content of type II colagen (P< 0.05). MMP-3 concentration and the difference value before and after experiment were gradualy decreased in DEF, D+L, D+H, and control groups in sequence(P< 0.05). These findings demonstrate that MMP-3 inhibitor I accelerates the early-stage repair of ful-thickness articular cartilage defects in the knee of rats.

Objective To analyze the molecular characteristics and prognosis in acute myeloid leukemia patients with AML1/ETO.Methods The clinical data of 63 cases of acute myeloid leukemia (AML) patients with AML1/ETO positive were analyzed retrospectively.56 cases of AML patients with AML1/ETO negative in the same period were analyzed as control.Characteristics in morphology,immunology,cytogenetics,molecular biology and the clinical effects of treatment were studied and analyzed.Results M2a was 57.12 ％ (36/63),M2b was 33.33 ％ (21/63) in AML with AML1/ETO.The percent of initial marrow blasts was 0.46±0.16.The positive rate of CD34,CD13,CD33,CD19,CD7 and CD56 was 67.21％,52.46 ％,40.98 ％,63.93 ％,4.92 ％ and 50.82 ％,respectively.The rate of t(8;21) translocation was 82.54 ％.There was 4.76 ％ with additional chromosome abnormality,three cases with EV1 1and one case with MLL/AT9.The overall CR rate,the relapse rate,the 3-year and the 5-year overall survival rate was 71.43 ％,51.11％,(43.01±5.31) ％ and (32.79±3.81) ％,respectively.There was no significant difference compared with the control group (P ＞ 0.05).But extramedullary infiltration,the expression of CD56 and additional chromosome abnormality had statistical effects on overall survival (P ＜ 0.05).Conclusions There has unique characteristics in AML with AML1/ETO.The effects of treatment and the prognosis are affected by many factors,so the efficacy and prognosis of AML with AML1/ETO couldn' t just depend on AML1/ETO.

Objective To investigate the incidence and risk factors of surgical site infection ( SSI ) in patients with colorectal cancer .Methods Clinical data of patients with colorectal cancer undergoing surgical treatment in Jiaxing First Municipal People’ s Hospital from October 2011 to December 2014 were retrospectively reviewed.The gender, age, underlying diseases, smoking history, preventive medication, abdominal surgery history , type of surgery , preoperative levels of hemoglobin and albumin , use of laparoscopy, use of stapler, combined organ resection, TNM staging, American Society of Anesthesiologists ( ASA) score was documented .Multivariate logistic regression analysis was performed to identify the risk factors of SSI .Results A total of 773 patients were enrolled in the study , and SSI was observed in 144 cases (18.63%).Multivariate logistic regression analysis showed that use of laparoscopy ( OR =0.35, 95%CI:0.15-0.79,P <0.05), use of stapler (OR =0.59, 95% CI: 0.39-0.88,P <0.05) were protective factors for SSI, while diabetes (OR=2.11, 95% CI: 1.25-3.58,P<0.01), liver cirrhosis (OR=2.12,95%CI:1.18-3.79,P<0.05), ASA score (3-4 points) (OR=2.01,95%CI:1.20-3.58, P<0.01), combined organ resection (OR=2.17,95% CI:1.20-3.92,P<0.05), and anastomotic leak (OR=6.85, 95%CI:3.01-15.63,P<0.01) were risk factors for SSI.Conclusions The incidence of SSI is high in patients with colorectal cancer undergoing surgery .Use of laparoscopy and stapler may reduce the incidence of SSI .

Administration, Intravenous ; Angina Pectoris, Variant ; drug therapy ; Humans ; Male ; Middle Aged ; Solanaceous Alkaloids ; administration & dosage ; therapeutic use

OBJECTIVETo investigate the risk factors of intra-abdominal infection(IAI) after colorectal cancer surgery.

METHODSClinical and follow-up data of 773 colorectal cancer patients undergoing operation in our hospital from October 2011 to December 2014 were retrospectively analyzed. Patients were divided into intra-abdominal cavity infection group (110 cases, IAI group) and non intra-abdominal infection group(663 cases, non-IAI group). All the patients administered prophylactic antibiotics 30 minutes to 2 hours before operation. Univariate and multivariate analysis were performed to evaluate the risk factors of IAI.

RESULTSPreoperative factors associated with postoperative IAI included hepatic cirrhosis, kidney diseases, diabetes or other basic diseases, prophylactic use of drugs, hypoalbuminemia, anemia, intestinal obstruction, and American Society of Anesthesiologists (ASA) anesthetic grading score (all P<0.05). Postoperative factors associated with postoperative IAI included use of laparoscopy or stapler, united exenteration, existence of anastomotic fistula, time of drainage tube placement, operation time and tumor staging (all P<0.05). Multivariate logistic regression analysis showed that preoperative diabetes(OR=2.36, 95% CI:1.45 to 4.76, P<0.01), combined exenteration (OR=2.02, 95% CI:1.02 to 4.00, P<0.01), anastomotic leak (OR=4.41, 95% CI:1.77 to 10.99, P=0.001), operation time≥140 minutes (OR=2.88, 95% CI:1.78 to 4.67, P<0.01) and period of postoperative drainage≥10 days(OR=4.57, 95% CI:2.78 to 7.52, P<0.01) were independent risk factors of postoperative IAI, while the use of stapler was protective factor (OR=0.37, 95% CI: 0.23 to 0.60, P<0.01). Compared with prophylactic use of cephamycins plus metronidazole, cefuroxime plus metronidazole had a higher rate of IAI(OR=2.10, 95% CI:1.23 to 3.58, P=0.007).

CONCLUSIONSPrevention of postoperative IAI is required for colorectal cancer patients, particularly in those with preoperative diabetes, combined exenteration, anastomotic leak, operation time longer than 140 minutes and postoperative drainage period longer than 10 days. Preoperative use of cephamycins plus metronidazole has better efficacy in prevention of postoperative IAI.

Anastomotic Leak ; Colorectal Neoplasms ; surgery ; Digestive System Surgical Procedures ; adverse effects ; Drainage ; Humans ; Intestinal Obstruction ; Intraabdominal Infections ; epidemiology ; Laparoscopy ; Neoplasm Staging ; Postoperative Complications ; epidemiology ; Retrospective Studies ; Risk Factors