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 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 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 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.

Here we discuss whether N terminal sequencing is appropriate as one of the conventional control methods for monoclonal antibody products. We determined the N terminal sequences of two monoclonal antibody products targeting two antigens separately with both Edman degradation and mass peptide spectrometry. We also identified the characteristic peptide fragments with mass spectrometry. Furthermore, we analyzed their heterogeneity with ion exchange chromatography, capillary zone electrophoresis and Imaged Capillary Isoelectric Focusing. Edman degradation method showed that the N terminal 15 amino acids of heavy and light chains of the two monoclonal antibodies were identical. Peptide mass spectrometry demonstrated that T1 peptide fragments of heavy and light chains of the two antibodies were also the same. But in contrast, peptide mapping and the three analytical methods for heterogeneity analysis could effectively identify and differentiate the two antibodies. The N terminal sequences of two monoclonal antibodies are identical because the number of framework sequences of humanized or human monoclonal antibodies is relatively limited, so whether N terminal sequencing analysis could be regulated as one of the practical control methods should be carefully discussed. Our work also proves that the above analytical methods could combinatorially applied to the identification of monoclonal antibody products, and are more objective compared to N terminal sequencing.
Amino Acid Sequence ; Antibodies, Monoclonal ; isolation & purification ; Chromatography, Ion Exchange ; Humans ; Isoelectric Focusing ; Mass Spectrometry ; Peptide Mapping ; Peptides ; Sequence Analysis, Protein ; methods

Objective Recent studies reveal important roles of platelet P-seleetin on progression of atherosclerosis.In the present study,we examine the relation between Platelet P-selectin expression and severity of acute coronary syndromes.Methods One hundredand eighty-four consecutive patients with proven or clinically suspected acute coronary syndromes(ACS)were enrolled in the study.Level of P-selectin expression was determined bv flow cytometry.Platelet P-selectin level was expressed as the percentage of P-selectin positive platelet.Results The level of P-selectin was higher in patients with a single diseased coronary artery or multiplediseased arteries compared to thOSe with normal coronary arteries.P-selectin expression was significantly and positively correlatedwith angiographic Gensini score(r=0.323,P=0.029).Multiple regression analyses showed that the association of the percentage of P-selectin-positive platelets with ACS was independent of other clinical factors. Conclusions Platelet P-selectin is associated withseverity of acute coronary syndromes in patients with acute coronary syndromes.

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.

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.