Objective To study the relationship of matrix metalloproteinase 7(MMP7)expression to the infiltration and metastasis of malignant melanoma.Methods Various concentrations(10,20,30 μmol/L)of heparanase antisense oligodeoxynucleotide (ASODN)was used to transfect human malignant melanoma cell line A375.Then,the transfected cells were subcutaneously inoculated into the right back of nude mice.Six weeks after the inoculation,transplanted tumors were isolated from the mice,and the levels of MMP7 protein expression were assessed by SP immunohistochemistry.The expression rate of MMP7 was also examined by SP immunohistochemistry in A375 cells and in tissue samples from patients with malignant melanoma(n=30),iunctional nevus(n=30)and normal human controls(n=15).Results The expression rate of MMP-7 was 83.33%(25/30),6.67%(2/30)and 0(0/15)respectively in malignant melanoma,iunctional nevus and normal skin tissue samples;there was a significant difierence in the positivity rate of MMP-7 between the malignant melanoma group and iunctional nevus group (x2=35.62,P=0.000)as well as normal control group(x2=28.12,P=0.000),but not between the junctional nevus group and normal control group(P＞0.05).The expression of MMP-7 in transplanted tumor was decreased to a varying degreee by three levels of ASODN,and the inhibitory effects of 30 mol/L ASODN were the strongest(P＜0.05).MMP-7 protein was also highly expressed in A375 cells.Conclusions The expression of MMP-7 in cutaneous malignant melanoma is higher than that in iunctional nevus and normal skin tissue.Hpa-ASPDN significantly inhibits the expression of MMP-7 in transplanted tumors in nude mice.MMP-7 is highly expressed in A375 cells.

Objective To establish a matrine delivery system in vitreous is very important for the dynamic treatment of proliferative vitreoretinopathy(PVR) . Present study was to evaluate the efficacy of matrine polyactic acid microsphere(MAT-PLA-MS) in prevention of PVR. Methods The suspension of cultured fibroblasts was injected into vitreous cavity of 30 healthy adult New Zealand albino rabbits to induce PVR. Then the experimental rabbits were divided into 3 groups and 10 rabbits for each. The animals received intravitreal injection of 0.3 mL MAT-PLA-MS(4 mg) matrine in MAT-PLA-MS group. Free matrine normal sodium solution 0.3 mL(containing 2mg matrine) was injected in vitreous cavity in free matrine group. 0. 3 mL normal saline solution was injected into the vitreous of the left eyes and the equivalent volume of blank polyaetic acid microsphere(blank-PLA-MS) into the right eyes in control group. The changes of cornea, aqueous humor, lens, vitreous and fundus were examined and recorded by slit lamp biomicroscope, indirect ophthalmoscope, fundus color camera and B ultrasonogram on the 1st, 3rd, 7th, 14th, 21st, 28th and 35th day following injection of drug. The inhibition effect of matrine on PVR was evaluated according to Ryan' s grading criteria of PVR. Results On the 14th days after implantation of MAT-PLA-MS, the rate of retinal detachment was 60%, 10%, 5% and 60% in normal saline group, free matrine group, MAT-PLA-MS group and blank-PLA-MS group respectively. Statistically significant difference was found among normal saline group, blank-PLA-MS group, MAT-PLA-MS group and free matrine group(P ＜0. 05). On the 21st day after injection of fibroblasts, the morbidity of retinal detachment was 80%, 30%, 10% and 80% in normal saline group, free matrine group, MAT-PLA-MS group and blank-PLA-MS group respectively, showing a significant difference among different groups. On the 28th day, the incidence rate of retinal detachment was 90%, 50%, 15% and 90% respectively, presenting statistical difference among various groups (P ＜ 0. 05) as well as between free matrine group and MAT-PLA-MS group (P＜0. 05). On the 35th day, considerably difference also was seen in the morbidity of retinal detachment among various groups (90%, 60%, 15% and 90% respectively) (P＜0.05). Conclusion Implantation of MAT-PLA-M S into vitreous cavity can effectively inhibit the development of PVR induced by fibroblasts in rabbit model.

ObjectiveTo investigate the expressions ofheparinase,matrix metalloproteinase 2 (MMP2) and tissue inhibitor of metalloproteinase 2(TIMP2) in malignant melanoma lesions and their significance.MethodsSkin specimens were obtained from the lesions of 30 patients with malignant melanoma,30 patients with melanocytic nevus and the normal skin of 15 healthy controls.Immunohistochemical SP method was used to detect the protein expression of heparinase,MMP2 and TIMP2.ResultsThe malignant melanoma tissue specimens significantly differed from the melanocytic nevus and control tissue specimens in the expression rate of heparinase (63.33％ vs.6.67％ and 0.00,x2 =21.172,27.805,both P ＜ 0.01 ),MMP2 (70.00％ vs.13.33％ and 0.00,x2 =19.817,19.866,both P＜ 0.01) and TIMP2(60.00％ vs.6.67％ and 0.00,x2 =19.200,15.000,both P ＜ 0.01 ).ConclusionThe expression of heparinase,MMP2 and TIMP2 is significantly higher in malignant melanoma lesions than in melanocytic nevus lesions and normal skin tissue.

Object To study the expression of miRNA-146a in sepsis-induced acute lung injury (ALI) patients and its effect on the inflammation in mouse models.Methods miRNA-146a expression in peripheral blood was determined in sepsisinduced ALI patients and healthy volunteers by qRT-PCR.Sepsis-induced ALI mouse model were reproduced by LPS treatment and miRNA-146a mimic,miRNA-146a NC and miRNA-146a inhibitors were injected through trachea.The expressions of miRNA-146a,TNF-α,IL-1β and COX-2 mRNA were determined by qRT-PCR 24h after the intervention.Results The miRNA-146a expression in peripheral blood significantly increased in severe sepsis with ALI patients,compared with the control subjects.For mouse model,the expressions of TNF-o,IL-1β and COX-2 mRNA significantly decreased in miRNA-146a group,while increased in miRNA-146a inhibitor group compared with miRNA-146a NC group (P＜0.05).The TNF-α and IL-1β expressions significantly decreased in miRNA-146a inhibitor+COX-2 inhibitor group compared with miRNA-146a inhibitor group (P＜0.05).Conclusion MiR-146a treatment can effectively alleviate the lung inflammation in sepsis-induced ALI mice.

ObjectiveTo investigate the expressions of matrix metalloproteinase-2 (MMP2) and tissue inhibitor of MMP2 (TIMP2) in human malignant melanoma transplanted subcutaneously in nude mice.Methods A non-sense oligonucleotide (N-ODN) and an antisense oligodeoxynucleotide (ASODN)against heparanase mRNA were designed and synthesized.Cultured human A375 malignant melanoma cells were classified into 4 groups to be transfected with the N-ODN,ASODN of 10,20,30 μmol/L,respectively,via liposomes.The cells receiving no treatment served as the blank control group.Then,the transfected or untransfected cells were subcutaneously inoculated into nude mice.Six weeks after the transplantation,transplanted tumors were removed from the nude mice and subjected to immunohistochemical staining for the detection of MMP2 and TIMP2 protein expressions.ResultsThe protein expressions of MMP2 and TIMP2 were significantly lower in tumor tissue specimens from nude mice inoculated with ASODN-transfected A375 cells than in those with N-ODN-transfected cells and in those with untransfected cells(P ＜ 0.05),significantly different between the tumor tissue specimens from mice inoculated with A375 cells transfected with 10,20 and 30 μmol/L of ASODN (all P ＜ 0.05 ).ConclusionThe ASODN against heparinase displays a marked inhibitory effect on the expressions of MMP2 and TIMP2 proteins in malignant melanoma transplanted in nude mice.

Objective To evaluate efficacy and safety of fixed combination of nitrendipine and atenolol in treatment for patients with mild to moderate essential hypertension and their optimal dosage matching.Methods Totally,275 patients with essential hypertension were selcted from seven hospitals in Shanghai,Nanjing and Suzhou,China and randomized into five groups with same proportional probability in a double-blind,double-dummy,parallel active-controlled,multi-center clinical trial,receiving fixed combination of nitrendipine and atenolol at three different dosage matching (nitrendipine/atenolol 5/12.5 mg,5/10 mg,5/7.5 mg for groups 1,2 and 3),and nitrendipine (10 mg for group 4) or atenolol (25 mg for group 5),respectively for eight weeks.Results Mean reduction of diastolic blood pressure (DBP)was (17±7) mm Hg,(18±9) mm Hg and (17±7) mm Hg for groupl,2 and 3,respectively from the baseline,significantly greater than that in groups 4 and 5[(13±7) mm Hg and (12±6) mm Hg,respectively].Mean reduction of systolic blood pressure (SBP) was (21 ±11)mm Hg,(24±12) mm Hg,(23±11) mm Hg,(19±13) mm Hg and (18±9) mm Hg,respectively for the five groups from the baseline,and the reduction in group 2 was significantly greater than that in group 5,with an overall efficacy of 94.4%,98.1% and 88.2% for groups 1,2 and 3,respectively,all statistically higher than that in group 5 (71.4%) with P＜0.01,eight weeks after treatment.The ratio of patients with increased dose of antihypertensive agents in week 5 was lower in group 2 than that in the other four groups,with mild adverse reaction only,no obvious change in laboratory biochemical examinations,and no needs in special management.Conclusions Fixed combination of atenolol and nitrendipine with an optimal doses of 5 mg and 10 mg respepctively was effective and safe for mild and moderate hypertension with good tolerance.

Objective To explore the expression of the CENP-W in gliomas and investigate the effects of its invasion. Methods The expression level of the CENP-W in gliomas with varied pathologic grade were detected by immunohistochemical analysis,RealTime PCR,and Western Blotting. U251 cells were transfected with the specific siRNA to repress the CENP-W expression level. The invasion ability of U251 cells were examined by Transwell Chamber assay ,while RAS mRNA and protein levels were detected at the same time. Results The expression levels of the CENP-W in glioma tissues were significantly high and the CENP-W gene could enhance the invasion of U251 cells . The expression of RAS was down-regulated when the expression of CENP-W was repressed. Conclusion The CENP-W has an oncogenic role in human brain gliomas and may regulate the invasion of gliomas by adjusting the RAS signaling pathways.

Objective:To explore the necessity of the preventive use of antibiotics and the effects of age and operation time on the efficacy of inguinal hernia repair without tension,and to elucidate the clinical significance of the preventive application of prophylactic antibiotics in inguinal hernia repair without tension.Methods:A total of 228 patients with inguinal hernia repair without tension were selected,amomg them 42 cases with high infection factors were treated with antibiotics (treated group),and 186 cases were not treated with antibiotics(untreated group) during the preoperative period.The prophylactic antibiotics were given 30 min before surgery,and the conventional dose was not used more than 48 h after surgery.All the cases were treated with artificial repair materials for the procedure of inguinal hernial repair without tension.The age,highest body temperature,white blood cell count,operation time,hospitalization time,and postoperative body temperature of all the 228 cases were recorded and analyzed statistically.Results:The preoperative and postoperative white blood cell counts had significant differences between the patients<60 years and the patients≥60 years in untreated group (P<0.05);there were no significant differences in the operation time,hospitalization time and body temperature between the patients<60 years and the patients≥60 years in untreated group (P>0.05).Compared with the patients with the operation time>90 min,the white blood cell count and hospitalization time of the patients with the operation time ≤90 min were increased (P<0.05).There were no significant differences of the body temperature and age between the patients with the operation time >90 min and the patients with the operation time≤90 min (P>0.05).The white blood cell count,operation time,hospitalization time and postoperative body temperature of the patients between treated group and untreated group had no significant differences (P>0.05).Conclusion:The use of antibiotics in the high-risk patients and non-use of antibiotics in the majority of elective inguinal hernia repair without tension can ensure the safe and performability of the patients.

Objective:To investigate clinical efficacy of conformal sphincter preservation operation (CSPO) versus intersphincteric resection (ISR) in the treatment of low rectal cancer.Methods:The retrospective cohort study was conducted. The clinicopathological data of 183 patients with low rectal cancer who were admitted to two medical centers (117 in the Changhai Hospital of Naval Medical University and 66 in the Huashan Hospital of Fudan University) from August 2011 to April 2020 were collected. There were 110 males and 73 females, aged (57±11)years. Of 183 patients, 117 cases undergoing CSPO were allocated into CSPO group, and 66 cases undergoing ISR were allocated into ISR group, respectively. Observation indicators: (1) surgical situations of patients with low rectal cancer in the two groups; (2) postoperative complications of patients with low rectal cancer in the two groups; (3) follow-up; (4) influencing factors for prognosis of patients with low rectal cancer; (5) influencing factors for satisfaction with the anal function of patients with low rectal cancer. Follow-up was conducted using outpatient examination, questionnaire and telephone interview to determine local recurrence, distal metastasis, survival, stomal closure, satisfaction with the anal function of patients. Measurement data with normal distribution were represented as Mean±SD, and comparison between groups was analyzed using the t test. Measurement data with skewed distribution were represented as M (range). Count data were described as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test. Comparison of ordinal data was analyzed using the rank sum test.The Kaplan-Meier method was used to draw survival curves, and life table method was used to calculate survival rates. Log-rank test was used for survival analysis. Univariate analysis was performed using the linear regression. Variables with P<0.10 in the univariate linear regression analysis were included for multivariate analysis. Multivariate analysis was performed using the COX stepwise regression model and linear regression analysis. Results:(1) Surgical situations of patients with low rectal cancer in the two groups: cases with laparoscopic surgery, operation time, volume of intraoperative blood loss, distance from tumor to distal margin, cases with postoperative chemotherapy, duration of postoperative hospital stay were 44, (165±54)minutes, (142±101)mL, (0.6±0.4)cm, 76, (6.6±2.5)days for the CSPO group, respectively, versus 55, (268±101)minutes, (91±85)mL, (1.9±0.6)cm, 9, (7.9±4.7)days for the ISR group, showing significant differences between the two groups ( χ2=35.531, t=8.995, -3.437, -3.088, χ2=44.681, t=2.267, P<0.05). (2) Postoperative complications of patients with low rectal cancer in the two groups: 19 patients in the CSPO group had complications. There were 6 cases with grade Ⅰ complications, 12 cases with grade Ⅱ complications, 1 case with grade Ⅲb complication. Fourteen patients in the ISR group had complications. There were 4 cases with grade Ⅰ complications, 7 cases with grade Ⅱ complications, 1 case with grade Ⅲa complication, 2 cases with grade Ⅲb complications. There was no significant difference in the postoperative complications between the two groups ( χ2=0.706, P>0.05). Patients with complications in the two groups were improved after symptomatic and supportive treatment. There was no perioperative death in the postoperative 30 days of the two groups. (3) Follow-up: 183 patients received follow-up. Patients of the CSPO group and ISR group were followed up for (41±27)months and (37±19)months, respectively, showing no significant difference between the two groups ( t=-1.104, P>0.05). There were 2 cases with local recurrence and 9 cases with distal metastasis of the CSPO group, respectively, versus 3 cases and 4 cases of the ISR group, showing no significant difference between the two groups ( χ2=1.277, 0.170, P>0.05). The 3-year disease-free survival rate and 3-year total survival rate were 84.0% and 99.0% for the CSPO group, versus 88.6% and 92.8% for the ISR group, showing no significant difference between the two groups ( χ2=0.218, 0.002, P>0.05). The stomal closure rate was 92.16%(94/102) and 96.97%(64/66) for 102 patients of CSPO group and 66 patients of ISR group up to postoperative 12 months,respectively, showing no significant difference between the two groups ( χ2=1.658, P>0.05). Of the 8 cases without stomal closure in the CSPO group, 2 cases refused due to advanced age, 4 cases subjectively refused, and 2 cases were irreducible due to scar caused by radiotherapy. Two cases in the ISR group had no stomal closure including 1 case of postoperative liver metastasis and 1 case of subjective refusal. There were 92 and 61 patients followed up to 12 months after stomal closure, of which 75 cases and 38 cases completed questionnaires of satisfaction with the anal function. The satisfaction score with the anal function was 6.8±2.8 and 5.4±3.0 for CSPO group and ISR group, respectively, showing a significant difference between the two groups ( t=-2.542, P<0.05). Fifty-four cases in the CSPO group and 21 cases in the ISR group had satisfaction score with the anal function >5, showing no significant difference between the two groups ( χ2=3.165, P>0.05). (4) Influencing factors for prognosis of patients with low rectal cancer: results of COX stepwise regression analysis showed that gender and pT staging were independent influencing factors for disease-free survival rate of patients with low rectal cancer ( hazard ratio=2.883, 1.963, 95% confidence interval as 1.090 to 7.622, 1.129 to 3.413, P<0.05). Gender and pT staging were independent influencing factors for total survival rate of patients with low rectal cancer ( hazard ratio=10.963,3.187, 95% confidence interval as 1.292 to 93.063, 1.240 to 8.188, P<0.05). (5) Influencing factors for satisfaction with the anal function of patients with low rectal cancer: results of univariate analysis showed that surgical method and tumor differentiation degree were related factors for satisfaction with the anal function of patients with low rectal cancer (partial regression coefficient=1.464, -1.580, 95% confidence interval as 0.323 to 2.605, -2.950 to -0.209, P<0.05). Results of multivariate analysis showed that surgical method, tumor differentiation degree and preoperative radiotherapy were independent influencing factors for satisfaction with the anal function of patients with low rectal cancer (partial regression coefficient=1.637, -1.456, -1.668, 95% confidence interval as 0.485 to 2.788, -2.796 to -0.116, -2.888 to -0.447, P<0.05). Conclusion:Compared with ISR, CSPO can safely preserve the anus in the treatment of low rectal cancer, without increasing the incidence of postoperative complications, which can also guarantee the oncological safety and improve the postoperative anal function.

AIM: To explore the protective effects of sinomenine (SIN) on oxidative stress and pulmonary fibrosis and its relationship with the Kelch-like ECH-associated protein 1 (Keap1)/nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway. METHODS: MRC-5 cells were treated with hydrogen peroxide (H2O2) to establish the oxidative stress injury model, followed by administration with SIN. Cell viability was detected using the CCK-8 method. The biochemical kits were employed to measure malondialdehyde (MDA) content and superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) activities. The protein expression of Keap1 and Nrf2 was examined by western blot. Thirty SD rats were randomly divided into control group, bleomycin A5 (BLM) group and BLM + SIN group, with 10 animals in each group. Bleomycin A5 were intratracheally administered to the rats in BLM group and BLM+SIN group to establish the pulmonary fibrosis model. The rats in control group received the same volume of 9 g/L sodium chloride solution. The second day after model construction, the rats in BLM+SIN group were gavaged with SIN, while the rats in the other two groups were treated with 9 g/L sodium chloride solution. On day 28, all rats were sacrificed. Pulmonary tissue was isolated, and HE and Masson staining was performed to observe the pathological changes. The MDA content and SOD, GSH-Px and CAT activities in pulmonary tissue were evaluated. Western blot was used to assay pulmonary tissues Keap1 and Nrf2 protein expression. RESULTS: When compared with H2O2 group, SIN treatment increased cell viability, decreased MDA content, elevated SOD, GSH-Px and CAT activities, down-regulated Keap1 expression, and promoted nuclear translocation of Nrf2 in MRC-5 cells. In comparison with BLM group, administration of SIN decreased alveolitis and pulmonary fibrosis pathological changes and scores as well as pulmonary tissue MDA content, enhanced pulmonary tissues SOD, GSH-Px and CAT activities, down-regulated pulmonary tissues Keap1 expression, and raised Nrf2 levels in the nucleus. CONCLUSION: SIN alleviates oxidative stress and pulmonary fibrosis possibly by activating the Keap1/Nrf2 signaling pathway.