The efficacy, biodistribution and safety of an inhibin DNA vaccine delivered by attenuated Salmonella choleraesuis.

DNA vaccines, the third-generation vaccines, were extensively studied. The attenuated Salmonella choleraesuis (S. choleraesuis) was widely focused as a carrier to deliver DNA vaccines in the chromosome-plasmid balanced-lethal system. The efficacy of inhibin DNA vaccine delivered by attenuated S. choleraesuis was proved in mice and cows in our previous studies. In this study, the efficacy of inhibin DNA vaccine was confirmed in rhesus monkeys. To further study the biodistribution and safety, the mice were immunized under laboratory conditions. The results of the rhesus monkeys showed the plasma IgA and IgG titres against inhibin were elevated, and the oestradiol (E2 ) and progesterone (P4 ) levels were increased with immunizing inhibin DNA vaccine. The biodistribution and safety assessment displayed the body weight, pathological change and haematology indexes where there is no significant difference between vaccinated mice and control. And the genomics analysis showed there was no integration of the inhibin gene into the mouse genome 2 months after immunization. This study indicated the inhibin DNA vaccine delivered by attenuated S. choleraesuis was safe. And this vaccine was a potential means to improve their reproductive traits in primates and other animals.

Monitoring menstrual cycle, gestation and lactation by measuring urinary oestradiol and progesterone in the captive golden snub-nosed monkey (Rhinopithecus roxellanae).

The golden snub-nosed monkey is an endangered species and study of its reproductive physiology is crucial for the species' breeding programs. Urine samples (770) from 5 mature female golden snub-nosed monkeys were collected in the Shengnongjia Nature Reserve between October 2013 and December 2014 to monitor their menstrual cycle, gestation, and lactation. The concentrations of oestradiol (E2) and progesterone (P4) in the samples were measured by Chemiluminescent Microparticle Immunoassay (CMIA), and the hormone concentrations were indexed to creatinine levels to compensate for differences in water content. The results showed that the E2 and P4 levels during the breeding season were significantly higher than those during the non-breeding season (P<0.01). The length of the menstrual cycle during the breeding season was 24.29±0.71days (mean±SEM) with a follicular cycle of 8.33±0.62days and luteal cycle of 15.27±0.83days. In addition, the levels of E2 and P4 began to rise on day 14 and day 10 after conception and remained at a high level until parturition. However, the E2 and P4 levels during lactation were lower than those during gestation (P<0.01). In summary, this study extends our knowledge regarding the basic reproductive physiology of golden snub-nosed monkeys, which could play an important role in the expansion of this species' population.

[Effects of sophoricoside and genistein on biological characteristics of osteoblasts].

OBJECTIVE: To observe the effects of sophoricoside and genistein on the biological characteristics of osteoblasts. METHODS: Osteoblasts were isolated from parietal bone of the newborn SD rats (less than 24 hours) and cultured in minimum essential medium (MEM). The effects of different concentrations of sophoricoside and genistein (10, 1 and 0.1 micromol/L) on the bone formation of osteoblasts in vitro were observed by cell proliferation rate. Differentiation of osteoblasts was evaluated by detecting the activity of alkaline phosphatase (ALP) using 4-nitrophenyl phosphate disodium salt (PNPP) method and by measuring the mineralized nodiform dyed by alizarin bordeaux. And the activity of osteoprotegerin (OPG) promoters of human beings was tested by using gene cloning and reporter gene. RESULTS: Sophoricoside at 1 and 0.1 micromol/L could promote the proliferation of osteoblasts (vs control group, P < 0.05). Genistein at 10 micromol/L could inhibit the cell proliferation, while genistein at 0.1 micromol/L showed promotion to cell proliferation (vs control group, P < 0.05). The activity of ALP of osteoblasts was increased by sophoricoside at concentrations from 0.1 micromol/L to 10 micromol/L, but genistein only at concentration of 0.1 micromol/L could make it. The total areas of mineralization of the osteoblasts in 10, 1 and 0.1 micromol/L sophoricoside groups increased by 73%, 138.6% and 114.3% as compared with the control group, respectively, but those in different concentrations of genistein groups decreased. The activity of LacZ was increased by sophoricoside at concentrations from 0.1 micromol/L to 10 micromol/L, as well as by genistein at concentration of 0.1 micromol/L. CONCLUSION: Sophoricoside can stimulate the proliferation, differentiation and mineralization of osteoblasts. And it shows no inhibition to bone formation of osteoblasts compared with genistein. Sophoricoside can also increase the activity of osteoprotegerin promoter and is more effective than genistein.

[Evaluation of hepatocellular function influenced by Chinese drug Dahuang Zhechong pill].

OBJECTIVE: To evaluate the influence of Chinese drug Dahuang Zhechong pill on the hepatocellular function. METHOD: Thirty-seven patients with hepatocirrhosis and twelve normal controls were performed the hepatobiliary scintgraphy with Tc-99m labeled ethylene hepatobiliary iminodiacetic acid (99 mTc-EHIDA), and the biochemical examination of hepatic function. There was 19 cases repeated the imaging after 6 months treated with chineses drug. By the three compartmental model configurations, the function parameters of hepatocellular extraction and excretion were calculated. RESULT: In the hepatocirrhosis groups, the hepatocellular uptake peak time and mean residence index were higher than those in normal controls (P < 0.01). Compared to normal controls, the uptake index, uptake speed index and descendent speed index were decreased markedly (P < 0.05). After treatment for 6 months with Chinese drug, the level of serum transaminase, globulin and bilirubin was lower than that before treatment. The uptake peak time and mean residence index decreased notably after treatment for 6 months (P < 0.01), and the uptake index increased, (P < 0.05). CONCLUSION: Chinese drug Dahuang Zhechong pill may improve the hepatocellular function and liver function status in patients with hepatocirrhosis.

[Effect of antibody-targeted chemotherapy with pingyangmycin on prostate cancer cells in vitro].

OBJECTIVE: To investigate the effect of antibody-targeted chemotherapy against human prostate cancer LNCaP cells in vitro. METHODS: The monoclonal antibody 7E11C5.3 against human prostate cancer was conjugated to pingyangmycin (PYM), mediated by dextran T-40, and the immunoreactivity of 7E11C5.3 was determined by indirect enzyme-linked immunosorbent assay. The bacteriostatic activity of the conjugate was determined using TTC assay, and its cytotoxicity against LNCaP cells was determined by MTT assay. RESULTS: The 7E11C5.3:PYM molar ratio was l:54 in the conjugate, and the immunoreactivity of 7E11C5.3 was decreased by approximately 10% to 20% after conjugation. The bacteriostatic activity of conjugated PYM was 25% of that of free PYM. The 50% inhibitory doses (IC50) of 7E11C5.3-PYM conjugate and free PYM against the in vitro cultured LNCaP cells were 9.41-/+1.98 microg/ml and 29.92-/+7.88 microg/ml, respectively. CONCLUSION: 7E11C5.3-PYM conjugate displays stronger cytotoxicity against anti-prostate cancer effects than free PYM.

[Analysis of HLA-DRB1 allele polymorphism in the Fujian Han nationality population of China].

OBJECTIVE: To investigate the HLA-DRB1 allele polymorphism in Fujian Han nationality population. METHODS: Six hundred and twenty individual samples collected from unrelated Fujian Han population were subjected to genotyping using oligonucleotide microarray technique. And the allele frequencies of HLA-DRB1 were calculated and compared with other populations. RESULTS: Fourteen HLA-DRB1 alleles of Fujian Han population were detected. The gene frequencies ordered from high to low were HLA-DRB1*9, 12, 15, 4, 8 respectively. CONCLUSION: The HLA-DRB1 distribution of Fujian Han population shares some genetic characters with southern Chinese Han populations, but these characters differ from northern Chinese populations.

[Retrospective study of the relationship of height difference and bone density of postmenopausal women].

OBJECTIVE: To analyze the relationship of the height difference and bone density (BD) of premenopausal and postmenopausal women. METHODS: The height values of 191 premenopausal and postmenopausal women were recorded, and the BD values of lumbar vertebrae and hip were detected by double energy X-ray BD detector. RESULTS: The lower the height of the postmenopausal women, the less the BD value. The BD of lumbar vertebrae dropped 0.025 5 g/cm(2) with each 2 cm of the shortened height, and the BD of hip joint dropped 0.029 2 g/cm(2). The shortened value in postmenopausal women with osteoporosis was statistically greater than that in postmenopausal women without osteoporosis. CONCLUSION: The BD of the postmenopausal women can be estimated by the calculation of their shortened height value.

[Effect of Sophoricoside on histomorphology of bone in ovariectomized rats].

OBJECTIVE: To evaluate the potential preventive effect of Sophoricoside on bone loss in ovariectomized rats. METHODS: Female SD rats (n=50, 6 months old) were either sham-operated (SHAM group, n=10) or ovariectomized (n=40). Three days after operation, ovariectomized rats were randomly assigned to groups as follows: 10 received Sophoricoside 4 mg.kg(-1).d(-1) (SL group), 10 received Sophoricoside 8 mg.kg(-1).d(-1) (SM group), 10 received Sophoricoside 16 mg.kg(-1).d(-1) (SH group) and 10 were untreated (OVX group). After Sophoricoside were given orally for one month, the histomorphometric parameters in the secondary spongiosa of proximal tibia and lumbar vertebrae were examined. RESULTS: Compared with OVX group, SM and SH two kinds of treatment caused 15.28% and 22.81% increment in percent trabecular area (BV/TV, %) in the secondary spongiosa of proximal tibia, 14.23% and 21.2% increment in lumbar vertebrae. Accordingly in these two groups, there was a significant decrease in trabecular separation (FLAW, microm), but increment in trabecular width (Tb. Th, microm) and conjunction points (JOINT). But in SL group, the preventive effect was not observed. CONCLUSION: Sophoricoside can be efficient in preventing ovariectomy-induced bone loss in rats.