Objective:To purify HLA-DR molecules. Methods: Anti-HLA-DR antibody L243 was purified and coupled with CNBr activated Sepharose 4B gel. Immunoaffinity column was used to purify HLA-DR molecules. Results:Twenty micrograms of HLA-DR molecules were isolated from about 5 g Epstein-Barr virus-transformed human B lymphoblastoid cell line RAJI lysates by affinity chromatography. The purified HLA-DR molecules existed in α/β heterodimers form and could bind to conformation-dependent antibody L243. These HLA-DR molecules were separated into two strands,α and β,by boiling denaturation. These results are the basis for studying MHC Ⅱ binding peptide motif and CD4＋ T cell epitopes of antigens in future.

In traditional oral practice, the presystemic interactions with gut microbiota is an important mechanism underlying the holistic health benefits of Chinese herbal medicines (CHMs), making the study of CHMs distinct from the research of Western medicines of which the systemic exposure (level in blood) is the starting point and the core. Gut microbial metabolism complements host metabolism in maintaining metabolic homeostasis of many biologically important endogenous molecules and the disposition of numerous exogenous compounds. Among them, the widely distributed gut bacterial β-glucuronidases (BGUSs) coordinate with host UDP-glucuronosyltransferases (UGTs) to play a role in the occurrence and intervention of diseases by affecting the glucuronidation homeostasis and altering the intestinal local and/or systemic exposure of endogenous compounds and xenobiotics. On one hand, many ingredients of CHMs undergo enterohepatic circulation; On the other hand, CHMs can act on BGUSs directly or indirectly change the distribution and function of BGUSs through reprogramming gut microbiome. The multiple interactions between BGUSs and CHMs may play an important role in the overall therapeutic benefits of CHMs. This work firstly summarizes the latest research progress on BGUSs; then the physiological, pathological and pharmacological significance of BGUSs are exemplified with representative endogenous and exogenous compounds from the aspects of nutrient utilization, metabolic homeostasis, and therapeutic response based on the varied substrate spectra of BGUSs; finally, the scattered data in literature were integrated to summarize the multiple interactions between BGUSs and CHMs, highlighting the important role of BGUSs in the holistic actions of CHMs.

α-HgS is the main component of traditional Chinese medicine cinnabar, while β-HgS is the main component of Tibetan medicine Zuotai. However, there was no comparative study on the dissolution and absorption in gastrointestinal tract and bioaccumulation in organs of mercury in Cinnabar, Zuotai, α-HgS and β-HgS. In this study, the dissolution process of the four compounds in the human gastrointestinal tract was simulated to determine the mercury dissolutions and compare the mercury dissolution of different medicines and the dissolution-promoting capacity of different solutions. To explore the absorption and bioaccumulation of cinnabar and Zuotai in organisms, mice were orally administered with clinical equivalent doses cinnabar and Zuotai. Meanwhile, a group of mice was given α-HgS and β-HgS with the equivalent mercury with cinnabar, while another group was given β-HgS and HgCl2 with the equivalent mercury with Zuotai. The mercury absorption and bioaccumulation capacities of different medicines in mice and their mercury bioaccumulation in different tissues and organs were compared. The experimental results showed a high mercury dissolutions of Zuotai in artificial gastrointestinal fluid, which was followed by β-HgS, cinnabar and α-HgS. As for the mercury absorption and bioaccumulation in mice, HgCl2 was the highest, β-HgS was the next, and a-HgS was slightly higher than cinnabar. The organs with the mercury bioaccumulation from high to low were kidney, liver and brain. This study is close to clinical practices and can provide reference for the clinical safe medication as well as a study model for the safety evaluation on heavy metal-containing medicines by observing the mercury dissolution, absorption, distribution and accumulation of mercury-containing medicines cinnabar and zuotai.
Animals ; Brain ; metabolism ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Gastrointestinal Tract ; metabolism ; Kidney ; metabolism ; Liver ; metabolism ; Male ; Mercury ; chemistry ; pharmacokinetics ; Mercury Compounds ; chemistry ; pharmacokinetics ; Mice ; Solubility

BACKGROUNDThe morbidity and mortality of prostate cancer have been increasing rapidly in recent China. There were few studies investigating prostate-specific antigen (PSA) values ranges in the healthy Chinese population. We performed this study to determine the distribution of serum PSA in a large healthy Chinese population.

METHODSFrom January 2001 to May 2008, 11 150 healthy Chinese men aged 30 - 79 years came to our hospital for routine health check-up. All subjects without a previous diagnosis of prostate cancer, a history of prostate surgery, or urogenital tract infection were proposed to undergo systematic serum PSA measurement and digital rectal examination (DRE). Men with normal DRE and PSA ≤ 4.0 ng/ml and those PSA > 4.0 ng/ml or abnormal DRE but without adverse findings on prostate biopsy were included (n = 9358). Age and serum PSA concentration were recorded and correlated through Logistic regression analysis.

RESULTSThe 95th percentile serum PSA concentration was 1.89 ng/ml for men aged 30 to 39 years, 2.19 ng/ml for men aged 40 to 49 years, 2.88 ng/ml for men aged 50 to 59 years, 4.42 ng/ml for men aged 60 to 69 years, and 6.52 ng/ml for men aged 70 to 79 years. The serum PSA concentration correlated with age (P < 0.0001) with an annual increase of 0.97% for men in 40 years, 1.58% for men in 50 years, 3.04% for men in 60 years, and 3.99% for men in 70 years.

CONCLUSIONSThe serum PSA level correlates directly with age in Chinese men older than 40 years, not in Chinese men younger than 40 years old. Chinese men have lower PSA level compared with white men above 60 years of age, not in those under 60 years of age.

Adult ; Age Factors ; Aged ; Asian Continental Ancestry Group ; Humans ; Male ; Middle Aged ; Prostate-Specific Antigen ; blood ; Prostatic Neoplasms ; blood ; epidemiology

AIM:To investigate the effects of exosomes secreted by pancreatic cancer cells on the viability and function of βcells and the possible mechanism .METHODS:ExoQuick-TC kit was used to extract exosomes in the super-natants of mouse pancreatic cancer Pan 02 and MPC-83 cells, and the extracted exosomes were identified by transmission electron microscopy.Fluorescence-labeled exosomes were incubated with mouse insulinoma MIN 6 cells for 48 h to detect whether exosomes secreted by pancreatic cancer cells were uptaken by MIN 6 cells.MTT and glucose-stimulated insulin se-cretion ( GSIS) assays were conducted to examine cell viability and insulin secretion of MIN 6 cells after incubating with ex-osomes.The expression of miR-204 and Bcl-2 mRNA in MIN6 cells was detected by qPCR .The protein expression of Bcl-2, Bax, caspase-3 and cytochrome C (Cyt-C) in MIN6 cells was determined by Western blot .RESULTS:The results of transmission electron microscopy showed that both Pan 02 cells and MPC-83 cells secreted exosomes , and Pan02 cells secre-ted more.The co-incubation results of fluorescence-labeled exosomes and MIN6 cells confirmed that MIN6 cells were able to ingest large amounts of exosomes secreted by pancreatic cancer cells .The results of MTT and GSIS assays showed that the viability and the level of high glucose-stimulated insulin secretion of MIN 6 cells in exosome treatment group significantly decreased compared with nontreatment group (P<0.01).The results of qPCR showed that the exosomes secreted by pan-creatic cancer cells were rich in miR-204, and the mRNA expression of Bcl-2 in MIN6 cells was significantly down-regula-ted by exosome incubation ( P<0.01) .The results of Western blot showed that the protein expression of Bcl-2 in the MIN6 cells treated with exosomes was significantly down-regulated (P<0.05), and the protein levels of Bax, cleaved caspase-3 and Cyt-C in exosomes treatment group were significantly up-regulated ( P<0.01 ) .CONCLUSION: Pancreatic cancer cells secrete exosomes .The exosomes secreted by pancreatic cancer cells are ingested by βcells, and reduce the viability and insulin secretion of βcells.The mechanism may be related to the increase in exosomal miR-204 in the βcells.In-creasing miR-204 may inhibit the expression of Bcl-2 and promote the activation of mitochondrial apoptosis in βcells.

OBJECTIVETo perform a pharmacognostical study of the leaf of Uncaria hirsuta Havil.

METHODSThe specimens of Folium Uncariae Hirsutae were collected for studying its characteristics, microscopic appearance and thin-layer chromatography.

RESULTSThe leaf of Uncaria hirsuta Havil was characterized by numerous multicellular non-glandular hairs, 2 lines of palisade tissue, a diacytic type of stoma, and clustered crystals in its parenchyma. At least two kinds of alkaloids identical to the control were identified in the specimens.

CONCLUSIONThe results can be used as the evidence for identification, formulation of the quality-control standards as well as further utilization of Folium Uncariae Hirsutae.

Alkaloids ; isolation & purification ; Chromatography, Thin Layer ; methods ; Pharmacognosy ; methods ; Plant Leaves ; anatomy & histology ; chemistry ; Uncaria ; chemistry

In recent years, studies of large-scale systematic evaluation on clinical effectiveness of acupuncture were carried out in overseas. The literatures were conducted in Cochrane Library and overseas journals about systematic review of clinical effectiveness of acupuncture. The Cochrane Library contained a series of systematic reviews for the treatment of 67 kinds of diseases by acupuncture in 2009. Preliminary evaluations of clinical effectiveness of acupuncture on 37 kinds of disease were conducted. The results indicated that acupuncture therapy was effective for 7 kinds of disease, such as idiopathic headache, neck disorders, glaucoma, rheumatoid arthritis, chemotherapy-induced nausea or vomiting, primary dysmenorrhoea with TENS and knee osteoarthritis with TENS. However, these studies still need improved research designs and sufficient research evidence. The results also indicated that acupuncture was indecisive for the other 30 kinds of disease because of insufficient evidence. Through analysis, results of most systematic reviews indicated that there were no significant difference between therapeutic effects of acupuncture treatment and pseudo-acupuncture treatment. Effect of acupuncture treatment was equivalent to therapeutic effect of placebo. The likely reasons may be that some important clinical factors are disregarded in these researches, such as selection of acupoints, treatment with syndrome differentiation, the angel and depth of needle insertion, the proper time for treatment and so on. Therefore, the large-scale systematic evaluation on clinical effectiveness of acupuncture was criticized by acupuncturists. Thus, the pressing problem is to establish a rational evaluation system of clinical acupuncture. The suggestions are strengthening the research on diagnosis and treatment standard, strengthening the quality control of clinical acupuncture and establishing sound acupuncture control group and placebo acupuncture group. The basic researches on the relationship between diseases and acupoints need to be strengthened in order to explore the mechanism of acupoints reaction on diseases.
Acupuncture Therapy ; Clinical Trials as Topic ; Evaluation Studies as Topic ; Humans

Objective Using molecular methods to study the relationship between genotypes and serum resmtance of Neisseria gonorrhoeae in Wuhan area.Methods NG-mast and serum bactericidal assays at the molecular level were used to differentiate the 46 strains which were isolated from the outpatients of sexually transmitted disease clinics and the relationship between different genotypes while phenotypes was also studied.Results 80.43% of the 46 strains contained the island and we were able to define three dillerent combinations of genes in the isolates.Results from serum bactericidal assays showed that all 9 sac-4 stralns did not provide any serum resistance.Conclusion Different isolates carried clifferent gonococcal genetm islands(pathogenicity island)and certain phenotypes.There were no sobious relationship between sac-4 gene and serum resistance of Neisseria gonorrhoeae.

AIM: To investigate the radioprotective effect of adenine on irradiated lymphocytes and discover the possible mechanisms of protection. METHODS: Lymphocytes were pretreated for 12 h with adenine (0.001-0.1 μmol·L(-1)) and then exposed to 4 Gy radiation. Cell viability was observed by the MTS assay, apoptosis was detected by Annexin V-FITC/PI, DNA ladder, and caspase 3/7 activity. Caspase-9, Bax, and Bcl-2 gene expression was investigated by RT-PCR. RESULTS: Irradiation increased cell death and DNA fragmentation. Pretreatment with adenine significantly reversed this tendency. Furthermore, several anti-apoptotic characteristics of adenine were determined, including the ability to inhibit caspase 3/7, upregulate B-cell lymphoma (Bcl-2) and downregulate Bcl-2- associated X (Bax), capase-9 gene expression in 4 Gy-irradiated AHH-1 cells. CONCLUSION The results suggest that adenine had a radioprotective effect to inhibit apoptosis in a concentration dependent manner.
Adenine ; pharmacology ; Apoptosis ; drug effects ; radiation effects ; Caspase 9 ; genetics ; metabolism ; Cell Line ; Cell Survival ; drug effects ; radiation effects ; Gamma Rays ; Gene Expression ; drug effects ; radiation effects ; Humans ; Lymphocytes ; drug effects ; radiation effects ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Radiation-Protective Agents ; pharmacology ; bcl-2-Associated X Protein ; genetics ; metabolism

OBJECTIVESalmonella isolates recovered from retail meats that were collected in supermarkets and free markets in Xi'an and Yangling areas of Shaanxi province were studied to determine antibiotic susceptibility.

METHODAntimicrobial susceptibility to 14 antibiotics of 193 salmonella isolates were determined by using agar dilution method, which was recommended by National Committee of Clinical Laboratory Standard (NCCLS), and E.coli ATCC25922 and E.faecalis ATCC29212 as standard control strains.

RESULTSThe 44.6% of the salmonella isolates were resistant to sulfamethoxazole, followed by resistance to kanamycin (40.9%), tetracycline (37.8%), amoxicillin (26.9%), ampicillin (25.4%), gentamicin (23.3%) and chloramphenicol (21.8%). Some isolates also showed resistance to fluoroquinolones, the rates for ciprofloxacin, enrofloxacin, levofloxacin and gatifloxacin were 22.3%, 21.8%, 20.8% and 21.2%, respectively. 55 isolates (28.5%) were multidrug resistant (MDR) strains, 28 of 193 isolates (14.5%) could resist at least 13 antibiotics, 24 isolates (12.4%) were resistant to from 4 to 12 antibiotics.

CONCLUSIONSalmonella isolates recovered from retail meats in Xi'an district of Shaanxi province were seriously resistant to antimicrobials commonly used as human and veterinary medicine.

Animals ; Cattle ; Chickens ; Drug Resistance, Multiple, Bacterial ; Food Microbiology ; Goats ; Meat Products ; microbiology ; Salmonella ; drug effects ; isolation & purification ; Sheep ; Swine