The high performance liquid chromatography-fluorescence micelle assay (HPLC-FMA) method for the content determination of polysorbate 80 in monoclonal antibody drugs was validated to study its applicability and transferability between various laboratories, and the feasibility to be included in the Chinese Pharmacopoeia. Both J.T. Baker and Nanjing Well-sourced polysorbate 80 was used in the collaborative validation of polysorbate 80 content analysis in seven different laboratories. The results show that when the protein concentration was no more than 20 mg·mL^-1 and the concentration of polysorbate 80 ranged from 0.05 to 0.5 mg·mL^-1, the method had good specificity. The recovery rates of the spiked samples ranged from 92.20% to 117.70% for J.T.Baker and from 93.90% to 117.20% for Nanjing Well. The intra-laboratory precision (%RSD) was less than 4.30% for J.T. Baker, and less than 2.60% for Nanjing Well, while the overall precision was less than 5.45% for J.T. Baker, and less than 6.70% for Nanjing Well. The linear correlation coefficient was more than 0.98 for J.T. Baker and more than 0.99 for Nanjing Well. The results of the collaborative validation prove that the HPLC-FMA method has good accuracy, precision, linearity, and specificity, and could be used for release control analysis of polysorbate 80 content in monoclonal antibodies across different laboratories.

Taxol is one of the most potent anti-cancer agents, which is extracted from the plants of Taxus species. Isopentenyl diphosphate isomerase (IPI) catalyzes the reversible transformation between IPP and DMAPP, both of which are the general 5-carbon precursors for taxol biosynthesis. In the present study, a new gene encoding IPI was cloned from Taxus media (namely TmIPI with the GenBank Accession Number KP970677) for the first time. The full-length cDNA of TmIPI was 1 232 bps encoding a polypeptide with 233 amino acids, in which the conserved domain Nudix was found. Bioinformatic analysis indicated that the sequence of TmIPI was highly similar to those of other plant IPI proteins, and the phylogenetic analysis showed that there were two clades of plant IPI proteins, including IPIs of angiosperm plants and IPIs of gymnosperm plants. TmIPI belonged to the clade of gymnosperm plant IPIs, and this was consistent with the fact that Taxus media is a plant species of gymnosperm. Southern blotting analysis demonstrated that there was a gene family of IPI in Taxus media. Finally, functional verification was applied to identify the function of TmIPI. The results showed that biosynthesis of β-carotenoid was enhanced by overexpressing TmIPI in the engineered E. coli strain, and this suggested that TmIPI might be a key gene involved in isoprenoid/terpenoid biosynthesis.
Amino Acid Sequence ; Carbon-Carbon Double Bond Isomerases ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; Paclitaxel ; biosynthesis ; Phylogeny ; Plant Proteins ; genetics ; Taxus ; enzymology ; genetics

OBJECTIVETo compare the clinical effects of circumcision and the foreskin-deglove plus shaft-fix (FDSF) procedure in the treatment of phimosis or redundant prepuce in obese adult males (body mass index [BMI] ≥ 28 kg/m²).

METHODSForty-four obese adult men with phimosis or redundant prepuce underwent circumcision (n = 24) or FDSF (n = 20) according to their own wishes. The patients in the circumcision and FDSF groups were aged (26.38 ± 4.24) and (26.90 ± 3.14) years, with BMIs of (27.77 ± 0.77) and (28.07 ± 2.28) kg/m² and penis lengths of (3.51 ± 0.46) and (3.50 ± 0.59) cm, respectively. The operations were performed under local anesthesia with lidocaine plus ropivacaine mesylate.

RESULTSThe operation time of circumcision was (28.04 ± 2.65) min and that of FDSF was (45.45 ± 3.49) min. At 6 months after surgery, normal penile erection was found in all the patients, the penis length was significantly longer in the FDSF than in the circumcision group ([5.01 ± 0.73] vs [3.70 ± 0.47] cm) , and the rate of satisfaction with penile appearance was markedly higher in the former than in the latter group (3.25 ± 0.71 vs 2.83 ± 0.56).

CONCLUSIONThe foreskin-deglove plus shaft-fix procedure under local anesthesia with lidocaine and ropivacaine mesylate may achieve desirable penile erection and appearance in the treatment of phimosis or redundant prepuce in obese adult patients.

Adult ; Amides ; Anesthetics, Local ; Body Mass Index ; Circumcision, Male ; methods ; Foreskin ; abnormalities ; surgery ; Humans ; Lidocaine ; Male ; Mesylates ; Obesity ; complications ; Operative Time ; Penile Erection ; Penis ; abnormalities ; Phimosis ; surgery

Objective To establish a post?treatment prognostic score model for newly diagnosed metastatic nasopharyngeal carcinoma, and to investigate the feasibility of stratified therapy. Methods A total of 263 eligible patients with newly diagnosed metastatic nasopharyngeal carcinoma from 2002 to 2010 were enrolled as subjects. The primary tumor was treated with conventional radiotherapy, three?dimensional conformal radiotherapy, or intensity?modulated radiotherapy, and radiation areas included nasopharyngeal tumor and cervical lymphatic drainage region. The metastatic bone tumor was mainly treated with conventional external radiotherapy, while the metastatic liver or lung tumor was mainly treated with surgical resection, radiotherapy, or radiofrequency ablation. The first?line therapy for most of patients was cisplatin?based combination chemotherapy. Factors including the general characteristics, tumor status, and therapy for patients were involved in multivariate analysis, and a prognostic model was established based on the n value (HR=en ) of the prognostic factors. Results The factors influencing the overall survival (OS) in patients were a Karnofsky performance score (KPS) not higher than 70(P= 0?? 00), multiple organ metastases (P=0?? 00), combination with liver metastasis (P= 0?? 00), a number of metastases not less than 2(P= 0?? 00), a level of lactate dehydrogenase (LDH) higher than 245 IU/ L (P= 0?? 00), a number of chemotherapy cycles ranging between 1 and 3( P= 0?? 00), a poor response for metastatic tumor ( stable disease or progressive disease)(P= 0?? 00), and primary tumor not treated with radiotherapy (P= 0?? 01). Based on the prognostic score, patients were divided into low?risk group (0?1?? 5 points), intermediate?risk group (2?? 0?6?? 5 points), and high?risk group (≥7?? 0 points), and the 5?year OS rates in the three groups were 59?? 0%, 25?? 1%, and 0%, respectively. Conclusions The prognostic score model based on the KPS, serum level of LDH, multiple organ metastases, combination with liver metastasis, and number of metastases can effectively predict the survival in patients. Active treatment including at least 4 chemotherapy cycles and radiotherapy for primary tumor can prolong the survival time of patients in the low?and intermediate?risk groups. However, patients in the high?risk group were mainly treated with palliative radiotherapy due to no improvement in the survival by radiotherapy for primary tumor.

Objective To investigate the clinical states of enuresis children companied with spina bifida occulta(SBO)and study the efficient way of treatment.Methods The children with SBO were check out by X ray from a total of 121 children with bedwetting.Their parents were asked to complete the enuresis questionnaires.Urine routine test and B-ultrasound examination about kidney,bladder and ureter were also asked to be done.The clinic data of the 49 children were attained and analyzed.They were randomly divided into 2 groups,and given the controlled treatment.Group A[used 1-deamino-8-D-arginine vas-opressin(DDAVP)only] and group B(used DDAVP plus oxybutynin plus bladder training)treated for 12 weeks.Results There were totally 49 bedwetting children companied with SBO,and most of them(44 cases,89.8%)were severe type(bedwetting times≥7 times/week).Some of them coexisted with frequency,urgency,gentle urgency incontinence and microscopic hematuria(22 cases).Thirty cases were found the functional bladder capacity(FBC)decrease by B-ultrasound.The cure rates were 58.3%(group A)and 88.0%(group B)respectively.The relapse rates were 36.8%(group A)and 12.5%(group B)respectively after stopping treatment for 3 months.Conclusions SBO accounts for considerably higher rate in enuretic children.It might cause the disability of bladder function.The treatment plan with DDAVP plus oxybutynin plus bladder training can not only increase the cure rate but also lower the relapse rate.

To set standards for histomorphological studies on Lysimachia fortunei, an efficacious and widely applied folk medicine in this study, in order to develop its resources. Its species were identified by observing plant morphology and herbs appearance characters, preparing slices with routine methods and defining structural characters. According to the results of morphologic observation, leaves, stamen and pistil of this plant were different from the descriptions in Flora of China. The whole herb can be used in medicines, mainly including rhizomes, stems and leaves. According to the findings in the first study on microscopic structures, its rhizomes, stems and leaves were characteristic and worth identifying. The transaction tissue structures of rhizomes and stems were under developed and contained endodermis, secretory structures; Stems had sclerenchymata of different shapes of sclereids; Leaves were bifacial and had vascular bundles under midribs, which were surrounded by parenchymal sheathes. On the surface of leaves, stomata, glandular hairs and keratin lines were morphologically different in upper and lower epidermis. The herbal power had glandular hairs, sclereids and vessels. In conclusion, herbs of L. fortunei can be identified by the above histomorphological characteristics, which lays a foundation for further development and application of L. fortunei.
Medicine, Traditional ; Plant Leaves ; anatomy & histology ; growth & development ; Plant Stems ; anatomy & histology ; growth & development ; Plants, Medicinal ; anatomy & histology ; growth & development ; Primulaceae ; anatomy & histology ; growth & development

Adult ; Antigens, Neoplasm ; genetics ; Carcinoma, Hepatocellular ; genetics ; pathology ; Female ; Humans ; Liver ; pathology ; Liver Neoplasms ; genetics ; pathology ; Male ; Middle Aged ; RNA, Messenger ; genetics

BACKGROUND: Human umbilical cord mesenchymal stem cells (hUC-MSCs) have low immunogenicity and it is unclear whether insulin producing cells (IPCs) that differentiate from hUC-MSCs have low immunogenicity. OBJECTIVE:To investigate the immunogenicity of IPCs differentiating from hUC-MSCs in vitro and after IPCs transplantation into the host. METHODS: (1) The hUC-MSCs were induced to differentiate into IPCs according to the modified scheme. Flow cytometry assay was used to detect the immunophenotype and apoptotic rate of IPCs in a cytotoxicity test. (2) Cell counting kit-8 was used to detect the proliferative capacity of human peripheral blood mononuclear cells in the one-way mixed lymphocyte assay. (3) The IPCs were then transplanted into the abdominal cavity and left renal capsule of mice, and then the infiltration of immune cells was detected by flow cytometry and immunohistochemistry. RESULTS AND CONCLUSION: The IPCs highly expressed HLA-ABC and lowly expressed HLA-DR, CD40 and CD80. The apoptosis rate of IPCs increased with the increase of pre-sensitized splenocytes in the cytotoxicity test. In the one-way mixed lymphocyte assay, IPCs inhibited the proliferation of human peripheral blood mononuclear cells when the target ratio was 10:1 and 50:1. After IPCs transplantation, the number of lymphocytes was increased in the transplanted site. In summary, our results show that IPCs that differentiate from hUC-MSCs maintain low immunogenicity in vitro,but have some immunogenicity after transplantation into the host due to microenvironment changes.

OBJECTIVETo study the protective effects and the possible mechanism of shengmai for injection(SM) against the experimental acute cardiogenic shock.

METHODThe experimental acute cardiogenic shock model was established by ligating the anterior descending cornonary in dogs. The effects of SM on cardiogenic shock were investigated by measuring the hemodynamics parameter, the activity of LDH, CK, SOD and the contents of MDA in blood serum.

RESULTIn the dogs treated with SM, the mean arterial pressure (MAP), heart rate (HR), left ventricular pressure (LVP), the maximum of its first derivative (+/- dp/dtmax), the cardiac output (CO) and the cardiac index (CI) were increased significantly. The left ventricular end diastolic pressure (LVEDP) and the peripheral vascular resistance (TPVR) were decreased significantly, the myocardial infarct size was redused observely. In addition, the activity of LDH, CK and the contents of MDA in serum were decreased significantly, however the activity of SOD was increased observely.

CONCLUSIONThe results indicate that SM has the protective effects on cardiogenic shock.

Animals ; Cardiac Output ; drug effects ; Cardiotonic Agents ; administration & dosage ; pharmacology ; therapeutic use ; Creatine Kinase ; blood ; Dogs ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Female ; Heart Rate ; drug effects ; Hemodynamics ; drug effects ; physiology ; Injections, Intravenous ; L-Lactate Dehydrogenase ; blood ; Male ; Malondialdehyde ; blood ; Myocardial Infarction ; pathology ; prevention & control ; Ophiopogon ; chemistry ; Panax ; chemistry ; Phytotherapy ; Plants, Medicinal ; chemistry ; Random Allocation ; Schisandra ; chemistry ; Shock, Cardiogenic ; blood ; physiopathology ; prevention & control ; Vascular Resistance ; drug effects

Objective To investigate the safety of rituximab combination chemotherapy in the treatment of B-cell non-Hodgkin' s lymphoma (B-NHL) complicated with hepatitis B virus (HBV) infection,and assess the incidence of HBV reactivation reduced by prophylactic lamivudine.Methods A retrospective study of HBV-related markers,HBV-DNA and liver function was performed before and after rituximabcontaining treatment in B-NHL patients.Thirty nine B-NHL patients with HBcAb(+)/HBsAb(-) were divided into prophylactic group (14 cases) and control group (25 cases).The incidences of HBV reactivation,functional damage of liver were measured.Results Among the 108 B-NHL patients who received rituximab combinatio nchemotherapy,15 (13.89 ％) were HBsAg (+) and 39 (36.11％) HBsAg (-) / HBcAb (+).Of the 15 HBsAg (+)patients,2 (13.3 ％) experienced reactivation of HBV.The prevalence of HBV reactivation was 7.7 ％(1/13) in patients who received prophylactic antiviral treatment and 50 ％ (1/2) in those who did not receivelamivudine.Among the 39 HBsAg (-) / HBcAb (+) patients,3 cases (7.7 ％) experienced reactivation of HBV.The prevalence of HBV reactivation was 0 in patients who receivcd prophylactic lamivudine treatment and 12 ％ (3/25) in those who did not receive this antiviral drug.Conclusion Prophylactic lamivudine before rituximab combination chemotherapy can reduce HBV reactivation obviously.