AIMTo explore the causes of the difference in spermatogenic suppression between responders and non-responders in Chinese men treated with levonorgestrel (LNG) implants plus testosterone undecanoate (TU) injectable.

METHODSThe 16 Chinese volunteers treated were divided into two groups in regard to the sperm count during the treatment period, 7 men in the responder group (Group R), including 6 azoospermia and one severe oligozoospermia, and the remaining 9 in the non-responder group (Group N), including 4 oligozoospermia and 5 with sperm counts greater than 20 x 10(6)/mL. The differences in serum profiles of FSH, LH, T, LNG and T/LH ratio were compared between the two groups and the correlation between the seminal fluid parameters and serum reproductive hormones was analyzed.

RESULTSThe serum FSH level was lower in Group R than that in Group N (P<0.05), while the serum LH and LNG levels were higher in Group R than those in Group N (P<0.05). The sperm density (P<0.01, r=0.235), motility(P<0.01, r=0.326) and vitality (P<0.01, r=0.219) showed significantly positive correlation with the serum FSH level.

CONCLUSIONThe blood LNG and T levels, the degree of FSH inhibition and/or the sensitivity of the pituitary-testis axis to exogenous steroids, as well as the individual spermatogenetic potential and the functional status of the Leydig cells may be factors bringing about individual differences in spermatogenic suppression in Chinese men treated with LNG and TU.

Adult ; Contraceptive Agents, Male ; administration & dosage ; Drug Implants ; Follicle Stimulating Hormone ; blood ; Humans ; Injections ; Levonorgestrel ; administration & dosage ; pharmacokinetics ; Leydig Cells ; physiology ; Luteinizing Hormone ; blood ; Male ; Sperm Count ; Spermatogenesis ; drug effects ; Testosterone ; administration & dosage ; analogs & derivatives ; blood

BACKGROUNDSericin peptide (SP) has shown a powerful anti-oxidant property in a host of studies. The present study was designed to investigate the possible protective effects of SP against alcohol-induced gastric lesions in mice and to explore the potential mechanisms.

METHODSAnimals were randomly divided into 5 groups: control, alcohol (56%, 14.2 ml/kg), SP-treated mice (0.2, 0.4, 0.8 g/kg). Mice were pretreated with SP before administering alcohol, the concentration of ethanol in serum and urine, the contents of malondialdehyde (MDA), glutathione (GSH) and the glutathione peroxidase (GSH-PX), catalase (CAT) and superoxide dismutase (SOD) activities in the gastric mucosa were measured, subsequently, the pathological evaluation of stomach was also observed.

RESULTSOf the animals pre-treated with SP (0.4, 0.8 g/kg), the concentration of ethanol in serum was significantly decreased, while increased in urine as compared to the alcohol-administered alone animals. Alcohol administration caused severe gastric damage as indicated by markedly increased MDA levels and decreased antioxidants, such as reduced GSH, GSH-PX and SOD in the gastric tissue while the CAT activity was not altered. On SP administration there was a reversal in these values towards normal. Histopathological studies confirmed the beneficial role of SP, which was in accordance with the biochemical parameters.

CONCLUSIONSSP could protect gastric mucosa from alcohol-induced mucosal injury. These gastroprotective effects might be due to increasing 'first-pass metabolism' in the stomach and hastening ethanol elimination directly through the urine. SP might also play an important role in the protection of the structure and function of gastric mitochondria, at least partly based on their anti-oxidant effect.

Amino Acids ; analysis ; Animals ; Cytoprotection ; Ethanol ; blood ; toxicity ; urine ; Gastric Mucosa ; drug effects ; pathology ; Glutathione ; metabolism ; Male ; Mice ; Mice, Inbred ICR ; Sericins ; analysis ; pharmacology ; Superoxide Dismutase ; metabolism

In order to enhance the antitumor efficacy of recombinant Newcastle disease virus, rNDV-IL15 was rescued in this study. Recombinant plasmid prNDV-IL15 was constructed, and BHK21 cells were transfected with the recombinant plasmid. Finally, the recombinant Newcastle disease virus rNDV-IL15 was successfully rescued. The growth curves of these two recombinant viruses were determined. Murine melanoma B16F10 cells were infected with rNDV-IL15 at MOI of 0.1, and the expression level of IL15 in the supernatant was detected by ELISA. The antitumor efficacy of rNDV-IL15 and rNDV was compared in vitro and in vivo. Results showed that prNDV-IL15 was constructed and recombinant virus rNDV-IL15 was successfully rescued. The growth curve of rNDV-IL15 showed that the growth of rNDV-IL15 had not been changed after insertion of IL15 gene. Results showed that there was high level of IL15 expression in the supernatant of rNDV-IL5-infected B16F10 cells (1 044.3 +/- 27.7 ng x mL(-1)). rNDV-IL15 and rNDV significantly inhibited the growth of B16F10 cells in vitro in a time-dependent manner. However, there was no significant difference between them. In animal experiments, rNDV-IL15 efficiently suppressed tumor growth in vivo when compared with rNDV, and the difference was statistically significant. The results suggested that rNDV-IL15 is a more effective antitumor agent.
Animals ; Body Weight ; Cell Line, Tumor ; Cell Proliferation ; Chick Embryo ; Cytotoxicity, Immunologic ; Female ; Genetic Therapy ; Interleukin-15 ; genetics ; metabolism ; Melanoma, Experimental ; pathology ; therapy ; Mice ; Neoplasm Transplantation ; Newcastle disease virus ; genetics ; Plasmids ; Recombinant Proteins ; genetics ; metabolism ; Transfection ; Tumor Burden

5-Flucytosine (5-FC) could be changed to 5-fluorouracil (5-FU) by cytosine deaminase (CD), the latter is able to kill cancer cells. However, there is no efficient method to deliver the CD gene into the tumor cells, which hampers the application of the suicide gene system. In this experiment, for the first time, the NDV has been utilized as a vector to deliver the CD gene into the cancer cells, the virus can infect the cancer cells specifically, replicate and assemble, while the cytosine deaminase is expressed. Then the CD converts the prodrug 5-FC into 5-FU to achieve the purpose of inhibiting tumor. Firstly, the whole genome of E. coli JM109 was extracted, and the CD gene was obtained by cloning method. Then the CD and IRES-EGFP were ligated into the pEE12.4 expression vector to become a recombinant pEE12.4IE-CD eukaryotic expression plasmid. The human liver cancer cells were transfected with the plasmid. The cells were treated with different concentrations of 5-FC, MTT method was used to determine the killing effect of CD/5-FC system on the human liver cancer cells. The cell deaths were 18.07%, 42.98% and 62.20% respectively when the concentrations of prodrug were at 10, 20 and 30 mmol x L(-1). In 5-FC acute toxicity experiment, Kunming mice were injected with different concentrations of 5-FC at intervals of 1:0.5. The LD50 of 5-FC through iv injection was determined by improved Karber's method, the LD50 was 507 mg x kg(-1) and the 95% confidence limit was 374-695 mg x kg(-1). According to the maximum LD0 dose of the LD50, the maximum safe dose was 200 mg x kg(-1). Body weight and clinic symptoms of the experimental animals were observed. These results laid the foundation to verify the antitumor effect and safety of CD/5-FC system in animal models. The CD gene was ligated into the NDV (rClone30) carrier, then the tumor-bearing animal was established to perform the tumor inhibiting experiment. The result showed that the recombinant rClone30-CD/5-FC system has a high antitumor activity in vivo. To summarize, CD gene has been cloned and its bioactivity has been confirmed in the mammalian cells. It is the first time in this study to utilize the recombinant NDV to deliver the CD gene into the tumor cells; our result proves the rClone30-CD/5-FC system is a potential method for cancer therapy.
Animals ; Antimetabolites, Antineoplastic ; metabolism ; pharmacology ; Cell Death ; drug effects ; Chick Embryo ; Cytosine Deaminase ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Flucytosine ; metabolism ; pharmacology ; Fluorouracil ; metabolism ; pharmacology ; Genetic Vectors ; Hep G2 Cells ; Humans ; Lethal Dose 50 ; Liver Neoplasms, Experimental ; pathology ; Mice ; Newcastle disease virus ; genetics ; Plasmids ; Recombinant Proteins ; genetics ; metabolism ; Transfection ; Tumor Burden ; drug effects

To investigate the cell-killing effect and its possible mechanism of rClone30-hDR5 in combination with TRAIL on human hepatic carcinoma (HCC) cell line, first of all, recombinant plasmid pee12.4-hDR5 was introduced into HepG2 cells by liposome transfection. After five rounds of screening by flow cytometry, HepG2 cells expressing high levels of DR5 on cell surface were isolated. The cytotoxicity of TRAIL to selected cells was higher than that of TRAIL to HepG2 cells by MTT method (P < 0.01). The result suggested that the cloned hDR5 gene had biological activity. MTT assay showed that, rClone30- hDR5 in combination with TRAIL more efficiently inhibited the tumor growth of HepG2 cells compared to rClone30-hDR5 or TRAIL in vitro. The results of Annexin V-FITC/PI staining and Quantitative Real-time PCR indicated that rClone30-hDR5 in combination with TRAIL significantly increased the mRNA levels of caspase 3 and caspase 8, and induced the apoptosis of tumor cells. HepG2 cells were infected with rClone30-hDR5 or rClone30 at MOI of 1. The expression of hDR5 on tumor surface increased significantly by rClone30-hDR5 compared to that by rClone30, which contributed to the sensitivity to TRAIL. In conclusion, rClone30-hDR5 in combination with TRAIL has potential application value in cancer treatment.
Apoptosis ; Carcinoma, Hepatocellular ; pathology ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Drug Synergism ; Hep G2 Cells ; Humans ; Liver Neoplasms ; pathology ; Real-Time Polymerase Chain Reaction ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; pharmacology ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology ; Transfection

This study aims to investigate the effects of fibroblast growth factor 21 (FGF-21) on learning and memory abilities and antioxidant capacity of D-galactose-induced aging mice. Kunming mice (37.1 +/- 0.62) g were randomly divided into normal control group, model group and FGF-21 high, medium and low dose groups (n = 8). Each group was injected in cervical part subcutaneously with D-galactose 180 mg x kg(-1) x d(-1) once a day for 8 weeks. At the same time, FGF-21-treated mice were administered with FGF-21 by giving subcutaneous injection in cervical part at the daily doses of 5, 2 and 1 mg x kg(-1) x d(-1). The normal control group was given with normal saline by subcutaneous injection in cervical part. At seventh week of the experiment, the learning and memory abilities of mice were determined by water maze and jumping stand tests. At the end of the experiment, the mice were sacrificed and the cells damage of hippocampus was observed by HE staining in each group. Reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT) and total antioxidant capacity (T-AOC) in the brain of mice were determined. The results showed that different doses of FGF-21 could reduce the time reaching the end (P < 0.01 or P < 0.05) and the number of touching blind side (P < 0.01 or P < 0.05) in the water maze comparing with the model group. It could also prolong the latency time (P < 0.05) and decrease the number of errors (P < 0.01 or P < 0.05) in the step down test. The result of HE staining showed that FGF-21 could significantly reduce brain cell damage in the hippocampus. The ROS and MDA levels of three different doses FGF-21 treatment group reduced significantly than that of the model group [(5.58 +/- 1.07), (7.78 +/- 1.92), (9.03 +/- 1.77) vs (12.75 +/- 2.02) pmol (DCF) x min(-1) x mg(-1), P < 0.01 or P < 0.05], [(2.92 +/- 0.71), (4.21 +/- 0.81), (4.41 +/- 0.97) vs (5.62 +/- 0.63) nmol x mg(-1) (protein), P < 0.01]. Comparing with the model group, the activities of SOD, GPx, CAT and T-AOC of the three different doses FGF-21 treatment groups were also improved in a dose-dependent manner. This study demonstrates that FGF-21 can ameliorate learning and memory abilities of D-galactose induced aging mice, improve the antioxidant abilities in brain tissue and delay brain aging. This finding provides a theoretical support for clinical application of FGF-21 as a novel therapeutics for preventing aging.
Aging ; drug effects ; Animals ; Antioxidants ; metabolism ; Brain ; drug effects ; Catalase ; metabolism ; Fibroblast Growth Factors ; pharmacology ; Galactose ; Glutathione Peroxidase ; metabolism ; Hippocampus ; drug effects ; Malondialdehyde ; metabolism ; Maze Learning ; drug effects ; Memory ; drug effects ; Mice ; Superoxide Dismutase ; metabolism

BACKGROUNDIslet transplantation is an effective way of reversing type I diabetes. However, islet transplantation is hampered by issues such as immune rejection and shortage of donor islets. Mesenchymal stem cells can differentiate into insulin-producing cells. However, the potential of human umbilical cord mesenchymal stem cells (huMSCs) to become insulin-producing cells remains undetermined.

METHODSWe isolated and induced cultured huMSCs under islet cell culture conditions. The response of huMSCs were monitored under an inverted phase contrast microscope. Immunocytochemical and immunofluorescence staining methods were used to measure insulin and glucagon protein levels. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to detect gene expression of human insulin and PDX-1. Dithizone-staining was employed to determine the zinc contents in huMSCs. Insulin secretion was also evaluated through radioimmunoassay.

RESULTSHuMSCs induced by nicotinamide and β-mercaptoethanol or by neurogenic differentiation 1 gene (NeuroD1) transfection gradually changed morphology from typically elongated fibroblast-shaped cells to round cells. They had a tendency to form clusters. Immunocytochemical studies showed positive expression of human insulin and glucagon in these cells in response to induction. RT-PCR experiments found that huMSCs expressed insulin and PDX-1 genes following induction and dithizone stained the cytoplasm of huMSCs a brownish red color after induction. Insulin secretion in induced huMSCs was significantly elevated compared with the control group (t = 6.183, P < 0.05).

CONCLUSIONSHuMSCs are able to differentiate into insulin-producing cells in vitro. The potential use of huMSCs in β cell replacement therapy of diabetes needs to be studied further.

Cell Differentiation ; genetics ; physiology ; Cells, Cultured ; Cellular Reprogramming ; genetics ; physiology ; Female ; Humans ; Immunohistochemistry ; Insulin-Secreting Cells ; cytology ; metabolism ; Mesenchymal Stromal Cells ; cytology ; Pregnancy ; Reverse Transcriptase Polymerase Chain Reaction ; Umbilical Cord ; cytology ; Wharton Jelly ; cytology

OBJECTIVETo investigate the clinicopathologic and immunophenotypic features of primary breast lymphoma (PBL) and to discuss the diagnosis of the tumor.

METHODSTwenty-one cases of PBL with follow up data were clinically reviewed. The histologic diagnosis of the tumor was based on the updated WHO Classification of tumors of hematopoietic and lymphoid tissues (2008). Immunohistochemistry was performed by SP method and antibodies selected were as follows: CD20, CD3epsilon, CD10, Bcl-6, MUM-1, CD5, Bcl-2, CD23, CD10, cyclin D1, CD43 and Ki67.

RESULTS(1) All 21 patients were female and the median age of patients was 48 years. The right and the left breasts were involved in 11 (52.4%) and 10 patients (47.6%), respectively. According to Ann Arbor staging system, 20 cases were stage I-II (95.2%), and the remaining case was stage IV (4.8%). For the international prognostic index (IPI), 19 cases were score 0-1, and 2 cases were score 2-3. For ECOG score, 19 cases were 0, and the remaining 2 cases were 1. (2) Histologically, all 21 cases (100%) were DLBCL. Immunohistochemically, the frequency of antigen expression was as follows: CD20 (100%), MUM-1 (14/21, 66.7%), bcl-6 (5/21, 23.8%), CD10 (0), bcl-2 (13/21, 61.9%), CD5 (1/21, 4.8%); Ki-67 index: 10 cases (47.6%) were less than 59%, with the expression of seven cases (33.3%) being 60% - 89%, and more than 90% in the remaining four cases (19.1%). The median Ki-67 index was 60%. All the cases were considered non germinal center B-cell-like type of DLBCL. (3) Follow-up data was available in 64% of the cases. One, two and five-year survival rates were 11 cases, 7 cases and 3 cases, respectively.

CONCLUSIONAll the cases of PBL in the current study were DLBCL, non germinal center B-cell-like type, and a diagnosis of PBL can only be established after excluding breast involvement in systemic lymphoma.

Adult ; Aged ; Antigens, CD20 ; metabolism ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Breast Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; Cyclophosphamide ; therapeutic use ; Doxorubicin ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Immunohistochemistry ; Interferon Regulatory Factors ; metabolism ; Ki-67 Antigen ; metabolism ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; metabolism ; pathology ; surgery ; Mastectomy, Radical ; Middle Aged ; Neoplasm Staging ; Prednisone ; therapeutic use ; Proto-Oncogene Proteins c-bcl-6 ; metabolism ; Retrospective Studies ; Survival Rate ; Vincristine ; therapeutic use ; Young Adult

OBJECTIVETo explore the surgical treatment of preoperative unsuspected gallbladder carcinoma (UGC) and its prognosis.

METHODSEighteen patients of UGC admitted in our hospital from January 1996 to December 2003 were analyzed retrospectively. The clinicopathological characters and surgical treatment of UGC patients were compared with the preoperative diagnosed gallbladder carcinoma (DGC) patients admitted in the same period. The cumulative survival rate of the patients received radical resection in two groups was analyzed by Kaplan-Meier method.

RESULTSThe serous layer was not invaded by tumor in 10 patients of UGC group (55.5%, 10/18), while the serous layer invaded and regional lymph node metastasis were detected in 39 patients of DGC group (90.7%, 39/43). Radical resection rate was 72.2% (13/18) in UGC group and 39.5% (17/43) in DGC group, and 5-year survival rates in those with radical resection were 54.6% and 23.5%, respectively (chi(L)(2) = 16.33, P < 0.01). Compared with the patients underwent palliative operation, the patients with radical resection has a longer median survival time (43.3 months vs 10.5 months, chi(L)(2) = 31.10, P < 0.01).

CONCLUSIONSThe prognosis of UGC is better than that of DGC generally. Reoperation for UGC should be performed as soon as possible, and the prognosis can be improved by radical resection.

Adult ; Aged ; Aged, 80 and over ; Female ; Gallbladder Neoplasms ; diagnosis ; mortality ; surgery ; Humans ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Survival Rate

OBJECTIVETo clone, express and characterize the capsid protein of human Norwalk virus Guangzhou strain NVgz01.

METHODSOn the basis of successful construction of full-genome clones and sequence analysis of human norovirus Guangzhou strain NVgz01, the full capsid gene was ligated into pET28a (+) for expression. After IPTG induction, the recombinant protein was purified through metal (Ni(2+)) chelating affinity chromatography. Western blotting and enzyme-linked immunosorbent assay (ELISA) were used to determine the antigenicity of the recombinant protein.

RESULTSThe recombinant capsid gene was overexpressed in E.coli, yielding the recombinant protein with relative molecular mass of 62x10(3) that was highly purified through metal (Ni(2+)) chelating affinity chromatography. IDEIA Norovirus Kit and immunoassay showed that the recombinant protein had good antigenicity.

CONCLUSIONThe capsid gene of norovirus Guangzhou strain has been cloned and expressed, which can be useful for developing diagnostic reagents or vaccine of norovirus.

Blotting, Western ; Capsid Proteins ; analysis ; biosynthesis ; genetics ; immunology ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; Humans ; Norwalk virus ; genetics ; Plasmids ; genetics