UNLABELLEDOBJECTIVE To explore the changes of bone gla protein (BGP) and tartrate resistant acid phosphatase (TRACP) in patients with stage 3 -4 chronic kidney disease (CKD) before and after treatment, to study their correlation with interleukin-17 (IL-17) and regulatory T cells (Treg), and the effects of Yishen Jiangzhuo Granule (YJG) on the bone metabolism.

METHODSFifty-three patients with stage 3-4 CKD were randomly assigned to the treatment group and the control group using random digit table. The following parameters in blood were detected: Treg (CD4+ CD25+ CD127lo) using tri-chrism fluorescent labeling by flow cytometry; levels of TRACP, BGP, and IL-17 by double antibody sandwich ELISA. The hemoglobin (HGB) content was detected using Beckman-Coulter heme/analysis. The urinary contents of creatinine (UCr) were determined using reversed HPLC. The blood contents of calcium (Ca), phosphate (P), blood urea nitrogen (BUN), serum creatinine (SCr), and plasma albumin (ALB) were determined using automatic biochemical analyzer. Then the calcium-phosphate (Ca x P) product was calculated on the basis of blood contents of Ca and P. The clearance rate of endogenous creatinine (CCr) was calculated on the basis of blood BUN and SCr contents.

RESULTS(1) There was no obvious change in CD4+ CD25+ CD127lo in the two groups before and after treatment (P > 0.05). Compared with before treatment in the same group, there were statistical difference in the levels of CD4+ and TRACP in the two groups, as well as the IL-17 level in the control group (P < 0.01, P < 0.05). But compared with the healthy group, statistical difference was shown in each index (except CD4+) (P < 0.01). Compared with the control group after treatment, there was no statistical difference in each index of the treatment group after treatment (P > 0.05). Compared with before treatment in the same group, the levels of Hb, ALB, and CCr increased (P < 0.05, P < 0.01), and the SCr level decreased in the two groups after treatment (P < 0.05). Compared with the control group after treatment, the SCr level decreased and the CCr level increased more obviously in the treatment group (P < 0.05). There was no correlation among the levels of IL-17, TRACP, BGP, and Treg between before and after treatment in the two groups.

CONCLUSIONSYJG could improve the kidney function and delay the progression of micro-inflammation of stage 3 -4 CKD patients. It could not improve the level of CD4+ CD25+ CD127lo. It also showed no effects on bone metabolism. The CD4+ T cells were differentiated to Th17 cells in stage 3-4 CKD patients. Their immunity was in a state of anergy but continually activated. The inflammatory factors in patients with stage 3-4 CKD play important roles in inducing the activation of osteoclasts.

Acid Phosphatase ; metabolism ; Adult ; Aged, 80 and over ; Bone and Bones ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Inflammation ; Interleukin-17 ; metabolism ; Isoenzymes ; metabolism ; Male ; Middle Aged ; Osteocalcin ; metabolism ; Renal Insufficiency, Chronic ; drug therapy ; immunology ; metabolism ; T-Lymphocytes, Regulatory ; immunology ; Tartrate-Resistant Acid Phosphatase

In the present study, we constructed a prokaryotic expression vector containing SOX4 protein encoding sequences. The GST-SOX4 soluble protein was expressed in Escherichia coli DH5alpha and purified by glutathione sepharose-4B. The purified recombinant protein was used to immunize Balb/C mice and the monoclonal antibody against SOX4 was prepared by using hybridoma technique. The titer of the antibody was determined as 1 x 10(5) by indirect ELISA. The specificity of the antibody was verified by Western blotting analysis. The monoclonal antibody specifically recognized the overexpressed exogenous SOX4 protein as well as endogenous SOX4 protein. The expression level of SOX4 protein in different cell lines and mouse tissues was detected by using the antibody. Differential expression of the protein was demonstrated by Western blotting. The data indicated that the antibody was specific. The antibody can be used as an important tool for further exploration of the role of SOX4 in tumorigenesis.
Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Glutathione Transferase ; genetics ; metabolism ; Humans ; Isoenzymes ; genetics ; metabolism ; Liver Neoplasms ; metabolism ; Lung Neoplasms ; metabolism ; Mice ; Mice, Inbred BALB C ; RNA, Messenger ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism ; SOXC Transcription Factors ; genetics ; immunology ; metabolism ; Tumor Cells, Cultured

Adult ; Biomarkers ; Humans ; Immunoglobulin A, Secretory ; analysis ; Isoenzymes ; analysis ; Male ; Occupational Diseases ; diagnosis ; immunology ; Saliva ; enzymology ; immunology ; Stress, Psychological ; diagnosis ; immunology

OBJECTIVEThe aim of this study was to investigate the mechanism(MDR) of multidrug resistance(MDR) of mucoepidermoid carcinoma in salivary gland.

METHODS40 cases of mucoepidermoid carcinoma in salivary gland were examined the MDR gene product P-glycoprotein using a monoclonal antibody JSB-1. And 10 of them were also investigated by detecting the expression of GST-pi. All the cases had not been accepted any therapy before the samples were collected.

RESULTS1. Positive expression of JSB-1 was observed in 27 of the 40 specimens. The positive expression was related not only with clinical stage, but also with differentiation degree. 2. The GST-pi positive expression was found in 9 of 10 cases. There was no significant different between the positive expression of JSB-1 and GST-pi.

CONCLUSIONJSB-1 and GST-pi play an important role in MDR of mucoepidermoid carcinoma.

ATP-Binding Cassette, Sub-Family B, Member 1 ; analysis ; Adolescent ; Adult ; Aged ; Antibodies, Monoclonal ; analysis ; Carcinoma, Mucoepidermoid ; immunology ; Child ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Female ; Genes, MDR ; Glutathione S-Transferase pi ; Glutathione Transferase ; analysis ; Humans ; Isoenzymes ; analysis ; Male ; Middle Aged ; Salivary Gland Neoplasms ; immunology ; Salivary Glands ; immunology

Lactate dehydrogenase-C4 isozyme has been widely studied as the sperm-specific antigen in the research of the contraceptive vaccine. Developmental and testis-specific expression of the LDH-C gene requires mechanisms for activation in germ cells and repression in somatic cells. LDH-C promoter, several transcription factors and methylation of CpG dinucleotides have been proved of governing action on LDH-C gene transcription. The strategy is to induce antibodies in the female reproductive tract against LDH-C antigens at a sufficient level to block fertilization. Chemically modified LDH-C4 offers a potential application in the induction of infertility of homologous species in marked contrast to native LDH-C4. In addition, advances are being made in researches on the male contraceptive vaccine and synthetic peptide immunocontraceptive.
Animals ; Humans ; Isoenzymes ; genetics ; immunology ; L-Lactate Dehydrogenase ; genetics ; immunology ; Male ; Mice ; Papio ; Rabbits ; Spermatozoa ; enzymology ; Transcription, Genetic ; Vaccines, Contraceptive ; immunology

The possible involvement of phospholipase C (PLC) in the regulation of insulin secretion is not clearly understood and neither its isozymes expressed nor cellular localization in the pancreatic islets is known. By using specific monoclonal antibodies, we have investigated the expression and localization of eight different PLC isozymes, beta1, beta2, beta3, beta4, gamma1, gamma2, delta1, and delta2, in the pancreatic islets of adult mice. Immunohistochemical analysis carried out on paraffin embedded sections showed a distinct pattern of expression for each of the PLC isozymes. In the central part of the islets containing beta cells, a high level of beta4 and moderate levels of beta3 and gamma1 were expressed, whereas PLC-beta1 and -gamma1 were abundantly expressed in the exocrine pancreas. These results demonstrated the heterogeneity in expression of the phospholipase C isozymes in pancreatic islets. It is conceivable that these isozymes are coupled to different receptors and perform selective tasks in the regulation of insulin secretion for glucose homeostasis.
Animal ; Antibodies, Monoclonal ; Glucagon/analysis ; Insulin/analysis ; Islets of Langerhans/cytology/*enzymology ; Isoenzymes/analysis/immunology ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Phospholipase C/*analysis/immunology

Excretion of urinary N-acetyl beta-D-glucosaminidase (NAG) and its isoenzyme patterns were studied in two groups of patients with rheumatoid arthritis (RA) and in normal control subjects. Urine samples were collected from 30 seropositive RA patients, 19 seronegative RA patients, and 15 normal healthy subjects. All the patients and normal subjects were assessed to have normal liver and kidney functions. A small portion of the urine sample was dialyzed against 0.01 M phosphate buffer, pH 7.0 and NAG activity was monitored. Mean +/- SD values of urinary NAG in seropositive RA patients, in seronegative RA patients and in normal healthy subjects were found to be 4.20 +/- 3.73 U/g creatinine, 2.96 +/- 2.11 U/gm creatinine, and 1.71 +/- 0.6 U/g creatinine, respectively. The mean urinary, NAG value in RA patients was found to be significantly higher (P < 0.05) in seropositive RA compared to the mean NAG value in normal healthy subjects and patients with seronegative RA when analyzed by one way ANOVA and Tukey-HSD test. The mean proportion of isoenzyme form B to isoenzyme form A in seropositive RA patients was also found to be significantly different (P < 0.05) from the mean proportion of these forms in normal healthy subjects and seronegative RA patients. There also appears to be a correlation between the concentration of urinary NAG and severity of the disease in seropositive RA.
Acetylglucosaminidase/urine* ; Adult ; Arthritis, Rheumatoid/urine* ; Arthritis, Rheumatoid/immunology ; Arthritis, Rheumatoid/enzymology* ; Chromatography, Liquid/methods ; Comparative Study ; Female ; Human ; Isoenzymes ; Male ; Predictive Value of Tests ; Severity of Illness Index

The clinical value of an enzyme-linked immunosorbent assay (ELISA) for the detection of anti-HBc IgM was evaluated by testing 202 sera from acute viral hepatitis B (AVHB), hepatitis B (HB), chronic hepatitis (CAH), chronic liver disease (CLD), cirrhosis, primary hepatoma, HBsAg carrier, acute viral hepatitis A (AVHA), hepatitis A (HA), non-A, non-B (NANB) hepatitis and miscellaneous conditions other than hepatic disease, and 19 additional various hepatic disease cases were examined for anti-delta. In clinical situations the accurate diagnosis of HB is not always possible and the differential diagnosis seems to be very important especially in making decisions of treatment and estimation of prognosis. In overall cases the highest positive rate of anti-HBc IgM was found in AVHB as shown as 74.3% (26/35) comparing to other conditions in which the positive rate was extremely low (2.1%). The anti-HBc IgM appeared to be highly specific to AVHB (83.9%) as compared to the other. The positive rate of HBsAg was high in AVHB, CAH and HBsAg carrier (100.0%) followed by CLD, cirrhosis and HB (up to 70.8%). The ALT activities and ALPalb fractions were significantly high in AVHB (p less than 0.005). The correlation between the positivity of anti-HBc IgM and highly abnormal ALT appeared be high. AVHB was confined mostly to 10-20 age group and the male to female ratio was about 6 to 1. Subgroup of AVHB II with positive anti-HBc IgM appeared to have a greater chance being positive for HBsAg and ALPalb. The S/N ratio of anti-HBc IgM was as high as 20 which was unique to AVHB.
Adolescent ; Adult ; Biological Markers ; Child ; Diagnosis, Differential ; Female ; Hepatitis/*diagnosis ; Hepatitis Antibodies/*analysis ; Hepatitis B/diagnosis/immunology ; Hepatitis B Antibodies/analysis ; Hepatitis Delta Virus/*immunology ; Humans ; *Immunoenzyme Techniques ; Immunoglobulin M/immunology ; Isoenzymes/immunology ; Male ; Middle Aged