Objective To discuss the clinical effect of combination therapy under the theory of "treating soft tissue for bone disease" on knee osteoarthritis.Methods The 160 cases were randomly divided into control group(80 cases) and experimental group(80 cases).The control group was treated by diclofenac sodium sustained release capsules(per os) and sodium hyaluronate(intra-joint injection).The experimental group was treated by traction,steaming and washing,massage,needle-knife therapy,manipulation on soft tissues for loosening the joint,and orthepedic insoles.The course of treatment was 3 weeks.The effect was evaluated according to Lysholm Score,knee movement and subjective satisfaction after the follow-up for 6 months.Results Of the 154 cases completed the study(control group 75 cases and experimental group 79 cases),the Lysholm Score and knee movement had a very significant improvement(P

Objective To investigate the influence of sperm morphology, sperm DNA fragmentation index and seminal plasma zinc on pregnancy outcome of in vitro fertilization and embryo transfer (IVF-ET).MethodsA total of 341 infertile couples underwent IVF-ET were selected from January 2016 to June 2016 in our hospital.Sperm morphology, sperm DNA fragmentation index and seminal plasma zinc level were compared according to pregnancy.ResultsIn 341 cases, 204 cases pregnancy and 137 cases of no pregnancy, with the pregnancy rate of 59.8%(204/341).Compared with the pregnancy group, the percentage of normal sperm percentage was low, the abnormal sperm index was higher in the non-pregnant group (P<0.05), the sperm DNA index was higher (P<0.05).The percentage of normal sperm, sperm DNA fragmentation index and seminal plasma zinc and pregnancy rate in sperm morphology were linear (P<0.05), And there was a negative correlation between abnormal sperm index and sperm DNA fragment index (P<0.05), and other indicators were positively correlated (P<0.05).ConclusionSperm morphology, sperm DNA fragmentation index and seminal plasma zinc levels may influence the outcome of IVF-ET.The above parameters can be used to predict pregnancy outcome of IVF-ET.

Objective To evaluate clinical features, diagnosis and treatment of anorectal malignant melanoma (ARMM).Methods The clinical data of 15 patients of ARMM in our hospital and 18 patients in the First Affiliated Hospital of China Medical University from 1990 to 2010 were reviewed.Twenty-five patients underwent curative surgical resection, 14 patients underwent abdominoperineal excision of the rectum (APR), and 11 patients underwent local excision (LE).Survival analysis was carried out.Fisher's exact test and Log-rank test was used to compare the effects of these two different surgical procedures.Results ARMM had a female predominance, the mean age was 22 -77(54.5 ± 7.6) years.The major clinical signs included hematochezia, anus pain.The misdiagnosis rate was 67% (22/33).The average tumor size was (3.5 ±1.7) cm.Thirty-one petients(94% ,31/33) had ARMM within 5 cm from anus margin.Mean survival time was (14.0 ± 6.5)months.The overall 1-,3-,and 5-year survival rates were 48% ,22% , and 10% , respectively.Local recurrence after curative LE was higher than APR (LE,64% vs APR, 21% , P = 0.049) , The overall 3-year disease-specific survival rates after curative LE was not significantly different from that of APR (LE, 28% vs APR ,31%, x2 = 0.268, P = 0.582).Conclusions Anorectal malignant melanoma has a high rate of misdiagnosis.Radical resection could not prolong the survival time significantly in anorectal malignent melanoma patients.

Objective To evaluate the efficiency and safety of transurethral plasmakinetic enucleation of prostate(PKEP) in the treatment of patients with benign prostate hyperplasia(BPH) after transrectal prostate biopsy(TRPB).Methods A total of 88 BPH patients who underwent PKEP in our hospital during Jan.2012 to May 2015 were retrospectively analyzed and followedup.38 patients underwent TRPB before PKEP were defined as TRPB group,and 50 patients underwent PKEP with no TRPB were defined as control group.The baseline data,perioperative data and postoperative follow-up outcomes were recorded and compared between the two groups.Results The mean age of the 88 patients were 69.7 years.Compared with control group,TRPB group showed that age was younger(t=2.62,P＜ 0.05)and prostate specific antigen(PSA) level was higher(t=13.64,P＜0.01).Operation duration was longer in TRPB group than in control group (93.6 ± 31.0) min vs.(77.9 ± 17.3) min (t =2.6 4,P ＜ 0.05).There were no significant differences in the preoperative data,blood loss,continuous bladder irrigation duration after operation and period of catheterization between two groups (all P ＞ 0.05).Trans-operative time was significantly shortened when the time interval between TRPB and PKEP is more than 4 weeks(P＜ 0.01),while blood loss was similar in the two groups(P＞ 0.0 5).There were no adverse events of blood transfusion,transurethral resection syndrome and injury of bladder or rectal in both two groups.There were no significant differences in maximum flow rate(Qmax),international prostate symptom score(IPSS) or quality of life(QOL) scores between the two groups at 3,6 and 12 months of follow up(all P＞ 0.05).Conclusions PKEP after TRPB is a safe and effective treatment for BPH patient.When the time interval between TRPB and PKEP is more than 4 weeks,the performing of PKEP operation would reduce the difficulty of operative procedure and increase the safety.

Benzoxazinoids (BXs) are important secondary metabolites in plants.There has been a wide range of attention and research of them because of their role in defensive and allelopathy.With the development of genomics and molecular biology,the BXs biosynthesis and other molecular areas research has made great progress.The BXs profile,the function of BXs,the genetic basis of BXs biosynthesis and expression regulation were briefly introduced.

Objective To investigate the role and potential molecular mechanism of the radiosensitivity of esophagus cancer cells.Methods miR-141 mimics or its negative control was transfected into esophagus cancercells KYSE-150,respectively.Radiosensitivity of esophagus cancer cells was determined by CCK-8,flow cytometry and colony formation assay.The expression level of miR-141 was determined by qRT-PCR.Western blot was used to detect the expressions of proliferation-related protein Ki67 and apoptosis-related proteins Bax and Bcl-2.Results After radiation,the expression of miR-141 was decreased in KYSE-150 cells in a dose-dependent manner (t =2.57-8.96,P ＜ 0.05).Upregulation of miR-141 significantly suppressed cell proliferation and colony formation and promoted apoptosis,indicating that overexpression of miR-141 enhanced the radiosensitivity of KYSE-150 cells(t =3.24,P ＜0.05).In addition,the miR-141 mimic significantly reduced the expressions of Ki67 and Bcl-2 protein (t =6.56,8.24,P ＜ 0.01) and inhibited the expression of Bax protein compared with miR-control group (t =3.24,P ＜ 0.01).Conclusions Over-expression of miR-141 enhances the radiosensitivity of esophagus cancer cells by regulating the expressions of proliferation-related protein Ki67 and apoptosisrelated proteins Bax and Bcl-2.

Objective To evaluated the risk factors of urethral recurrence ( UR) following radical cystectomy ( RC) in patients with bladder urothelial carcinoma.Methods The clinical data of 350 male patients who underwent RC between January 2005 and January 2013 were retrospectively analyzed.The mean age was 63 years (rang 46-76) years.176 cases had the history of non-muscle-invasive bladder cancer.15 cases were were found the tumor invasion into the prostatic urethral.The way of urinary diversion after RC included 172 cases were orthotopic neobladder, 90 cases were cutaneous diversion and 88 cases were ileal couduitin.331 cases underwent preoperation intravesical instillation.36 cases underwent systemic chemotherapy after operation.148 cases were found the multiple tumor lesions, which was more than 2 sites. The pathological stage was more than T2 satge in 189 cases.And 177 cases were diagnosed as high-grade urothelial carcinoma.Multivariate Cox regression analyses were used to evaluate the risk factors associated with the UR.Results There were 350 cases in this study, UR was observed in 28 cases ( 8%).On multivariate Cox regression analyses, previous history of NMIBC (HR=15.205,95%CI 3.718-62.180,P<0.001), prostate urethral involvement(HR=5.233,95%CI 1.106-24.754,P=0.037) and Non-orthotopic neobladder(HR=6.656,95%CI 1.840-24.077,P=0.004)which the operation of cutaneous diversion and ileal couduit , were independent risk factors of UR following RC.Intravesical instillation before operation ( HR =0.470, 95%CI 0.010-0.217, P <0.001 ) was the protective factor of the UR.Conclusions Previous history of NMIBC, prostatic urethral involvement and Non-orthotopic neobladder were independent risk factors of UR.Intravesical instillation before operation was protective factor of UR.Urethrectomy for patients with high risk factors and intravesical instillation before operation were important.

Objective To analyze the differences of size and charge heterogeneities between origi-nal humanized anti-TNF-αantibody and four similar biotherapeutic products ( SBP ) .Methods The size exclusion chromatography ( SEC-HPLC ) and weak cation exchange chromatography ( WCX-HPLC ) were used to analyze the size and charge heterogeneities , respectively.Carboxypeptidase B (CpB) treatment was employed to analyze the source of charge heterogeneity of the antibody products .Results Four SBPs showed the same pattern with the originator in SEC-HPLC, and no significant difference with the percentage of mono-mer was observed .The percentages of the aggregates of SBP-3 and SBP-4 were a little higher than those of the originator .The charge distribution of SBPs was significantly different from the originator ′s, especially in the basic region .The results from the samples treated with CpB indicated that the difference of charge distri -bution in the basic region might be caused by the C-terminal lysine variants .Conclusion Four SBPs showed similar size heterogeneity with the originator , but significant differences with charge heterogeneity were observed among them .The study suggested that more attention should be paid to the charge heterogene -ity analysis of the biosimilar products .

Objective To compare the capability of capillary electrophoresis-sodium dodecyl sul-fate ( CE-SDS) and size exclusion-high performance liquid chromatography ( SE-HPLC) for analysis of size heterogeneity of monoclonal antibody products .Methods The size heterogeneity of one humanized anti-VEGF monoclonal antibody was analyzed by using non-reduced and reduced CE-SDS, and conventional , de-natured and denatured reduced SE-HPLC.Results The percentage of aggregates detected by non-reduced CE-SDS (0.82%±0.01%) was equal to that by using denatured SE-HPLC (1.05%±0.02%), but it was significantly lower than that by using conventional SE-HPLC analysis (5.08%±0.10%).With regard to fragments analyzed with non-reduced antibodies, its percentage was (7.12±0.04)% measured by non-re-duced CE-SDS analysis that was significantly higher than that by conventional SE -HPLC analysis (0.02%± 0.01%) and denatured SE-HPLC analysis (0.62%±0.01%).Using reduced antibodies , the percentage of fragments was (3.19±0.50)%tested by reduced CE-SDS analysis that was significantly higher than that by using denatured reduced SE-HPLC analysis (0.07%±0.01%).Conclusion Conventional SE-HPLC was more objective than CE-SDS for content analysis of aggregates , as both the covalent and non-covalent forms of aggregates could be detected .Non-reduced CE-SDS could demonstrate the content of clips , while reduced CE-SDS showed the degraded fragments .Therefore, CE-SDS had an advantage over conventional SE-HPLC for content analysis of fragments .The use of the two analytical methods in combination provided solid techni-cal supports for the quality control of size heterogeneity of monoclonal antibodies .

The biological activity of ADCC by anti-CD20 monoclonal antibody was determined by BioGlo™ Luciferase Assay System using Jurkat/NFAT-luc+FcγRIIIa cell line as effector cell and WIL2-S cell line as target cell. The developed method was verified for specificity, precision and accuracy. Anti-CD20 monoclonal antibody showed a dose-response mode by the developed method, and the determination result complied with the following four-parameter equation: y = (A-D)/[1 + (X/C)(B)] + D. The optimized parameters of the method were determined including the antibodies diluted concentration (18,000 ng·mL(-1)), dilution rate (1:5), the ratio of effector cell and target cell (6:1), and induction time (6 h). The values of eight independent tests have passed a statistical test for curve regression analysis, linear or parallelism, which showed the method possessed good specificity. Four different dilute groups of recovery rates sample were determined for 3 times, and the result showed mean relative potencies of (44.39±3.93)%, (72.74±2.78)%, (128.28±7.01)% and (168.19±2.70)% respectively, with a variation coefficient of less than 10%, and the recoveries of (88.78±7.85)%, (96.99±3.70)%, (102.63±5.61)% and (112.12±1.80)% respectively. A novel reporter gene method for determination of biological activity of ADCC by anti-CD20 monoclonal antibody was successfully developed, which showed strong specificity, good reproducibility and high accuracy, and might be used routinely.