Properties of a Stable and Sustained-Release Formulation of Recombinant Human Parathyroid Hormone (rhPTH) with Chitosan and Silk Fibroin Microparticles.

BACKGROUND Parathyroid hormone (PTH) is required for the maintenance of normal bone physiology. This study describes the properties of a sustained-release formulation of recombinant human PTH (rhPTH) using chitosan and silk fibroin microparticles as carriers for drug delivery, developed using a spray-drying method. MATERIAL AND METHODS Chitosan, silk fibroin, and chitosan/silk fibroin microparticles loaded with rhPTH were studied with scanning electron microscopy (SEM) to estimate the particle size and surface morphology. The in vitro release of rhPTH was used to assess the developed formulation. The effect of the spray-drying process was assessed by powder X-ray diffraction (PXRD) of the microparticles. Quantification of the released rhPTH was performed by enzyme-linked immune sorbent assay (ELISA). Fourier-transform infrared spectroscopy (FTIR) was used to determine the differences in the absorption frequency of samples. RESULTS Surface morphology of the final formulation showed the absence of pure crystals of chitosan and silk fibroin in the final formulation and FTIR demonstrated electrostatic interactions between chitosan and silk fibroin, which was supported by PXRD. The chitosan/silk fibroin microparticles loaded with rhPTH showed an entrapment efficiency (EE) that ranged from 60.36-72.99% with a 50% rhPTH release profile at pH 7.5 in 24 hours. There was no particle aggregation in blood and little hemolysis, indicating stability of the rhPTH-loaded microparticles. CONCLUSIONS A silk fibroin/chitosan microparticle formulation loaded with rhPTH was shown to be stable and to provide sustained-release of rhPTH, supporting a potential role of this formulation in the treatment of bone diseases including osteoporosis and bone fracture.

[Content of neutrophil elastase in EPS and seminal plasma: A combined predictor in the diagnosis of type IIIA prostatitis with secondary infertility].

OBJECTIVE: To investigate the value of the content of neutrophil elastase (NE) in the expressed prostatic secretion (EPS) and seminal plasma (SP) as a combined predictor in the diagnosis of type IIIA prostatitis with secondary infertility. METHODS: This study included 62 fathers (group A) and 67 infertile men (group B), all with type IIIA prostatitis, and another 57 controls with no genitourinary tract disease (group C). We measured the NE contents in the EPS and SP, obtained the results of routine semen and EPS examinations and Chronic Prostatitis Symptom Index (CPSI), and calculated the ratio of EPS NE/SP NE by binary logistic regression analysis. RESULTS: The combined predictor of type IIIA prostatitis with secondary infertility was SP NE-2 × EPS NE. Among the 129 patients with type IIIA prostatitis, the combined predictor was correlated strongly negatively with the WBC count in the EPS (r = －0.914, P <0.01), negatively with CPSI, sperm concentration, sperm mobility and sperm viability (P <0.01), but not significantly with the WBC count or pH value in the SP (P >0.05). The mean values of the combined predictor in groups A, B, and C were －2 238 (95% CI: －2 595 to －2 054), －1 511 (95% CI: －1 778 to －1 307), and －148 (95% CI: －181 to －118), respectively, with statistically significant differences between the cases and controls as well as between groups A and B (P <0.01). The area under the ROC curve of the combined predictor for the diagnosis of type IIIA prostatitis with secondary infertility was 0.71 (P <0.001). CONCLUSIONS: The content of neutrophil elastase in the EPS combined with that in the seminal plasma contributes to the diagnosis of type IIIA prostatitis with secondary infertility, which is superior to either the neutrophil elastase content in the EPS or that in the seminal plasma used alone.

Survey of knowledge, attitude, and practice regarding reproductive health among urban men in China: a descriptive study.

There has been little focus on men's reproductive health (RH) in China. This descriptive study conducted in Yiling District, Yichang, China, surveyed male knowledge of sexual physiology and RH to assess levels of knowledge, attitudes and practices (KAPs) regarding prevention of sexually transmitted diseases (STDs). A total of 3933 men, aged 18-59 years (mean, 40.3 years), were recruited by cluster random sampling. They completed a questionnaire in the presence of an interviewer, with items related to subject characteristics, RH knowledge, and subjective symptoms of the reproductive system. Physical examination and reproductive system disease diagnosis were performed. Participants' occupations were predominantly skilled labor (80.5%). Nearly four-fifths (78.5%) respondents had at least one reproductive disease. Over half of respondents were aware of and declared a positive attitude about sexual physiology and safe sex, and 70% of them selected to visit a doctor when they had a reproductive disorder. However, only 41.9% believed human immunodeficiency virus/acquired immunodeficiency syndrome could be transmitted through breastfeeding, and 64.6% incorrectly thought they could avoid contracting STDs by cleaning their genitals after intercourse. In addition, 45% discriminated against and were unwilling to be friends with infected persons. Nearly 45% of those with a reproductive system disorder refused to discuss it with friends or family members. These results indicate that this cohort of Chinese men had a certain degree of KAP about RH, whereas some aspects require further public health education in the general population. It is necessary to disseminate accurate knowledge of STD risk in China based on sociodemographic characteristics.

Risk factors for cervical cancer in rural areas of Wuhan China: a matched case-control study.

Cervical cancer is a serious public health problem in developing countries. We investigated possible risk factors for cervical cancer in rural areas of Wuhan China using a matched case-control study with 33 women diagnosed with cervical cancer and 132 healthy women selected from the same area as matched controls. A questionnaire, which included questions about general demography conditions, environmental and genetic factors, the first sexual intercourse, first marriage age, age at first pregnancy, pregnancy first child's age, female personal health history, social psychological factors, dietary habits, smoking and alcohol status and other living habits was presented to all participants. At the same time, HPV infection of every participant was examined in laboratory testing. Results showed HPV infection (P<0.000, OR=23.4) and pregnancy first child's age (P<0.000, OR=13.1) to be risk factors for cervical cancer. Menopause (P=0.003, OR=0.073) was a protective factor against cervical cancer. However, there was no indication of associations of environmental (drinking water, insecticide, disinfectant) genetic (cancer family history), or life-style factors (smoking status, alcohol status, physical training, sleep quality), including dietary habits (intake of fruit and vegetable, meat, fried food, bean products and pickled food) or social psychological factors with cervical cancer. The results suggest that the risk of cervical cancer in Chinese rural women may be associated with HPV infection, menopause and the pregnancy first child's age.

[Study on the standard of health protection zone for petroleum processing industry].

OBJECTIVE: To establish health protection zone standards for petroleum processing industry. METHODS: The intensity of characteristic pollutants from fugitive emission were estimated by the inverse method of ground concentration through field survey and monitoring for representative petroleum processing industry, which was calculated health protection zone by using the model of atmospheric diffusion. RESULTS: Characteristic pollutant of fugitive emission source from petroleum processing industry was confirmed as hydrogen sulfide. When local average wind speed in the past five years was less than 2, 2-4 m/s and more than 4 m/s respectively and meanwhile the scale of petroleum processing industry was less than 8 million tons, the recommended value of health protection zone were 900, 800, 700 m respectively. Besides, when the scale of petroleum processing industry was more than 8 million tons and in the same wind speed level, the recommended value of health protection zone were 1200, 1000, 900 m respectively. CONCLUSIONS: Recommended value of health protection zone for petroleum processing industry was reasonable and feasible through revising and improving of the version of 1987's standard.

[Effect of NS-398 on cyclooxygenase-2 expression and proliferation of HepG2 cells].

OBJECTIVE: To investigate anticancer effect and molecular mechanism of N-[(Cyclohexyloxy)-4-nitrophenyl] methanesulfonamide on HepG2 cells in vitro. METHODS: HepG2 cells were treated with various concentrations (100, 200, 300, 400 micromol/L) of NS-398 [selective for cyclooxygenase 2 (COX-2) inhibition]. Cell growth was measured by MTT method, DNA fragmentation gel analysis was used to analyze the apoptosis cells, DNA ploidy and apoptotic cell percentage were examined by flow cytometry (FCM). PGE2 concentration was measured by radioimmunoassay method. The expressions of COX-2 were also examined by Western blot analysis. RESULTS: NS-398 inhibited HepG2 cell proliferation and induced apoptosis in a concentration-dependent manner. DNA ploidy analysis showed that S phase cells were significantly decreased and quiescent G1 phase was accumulated with NS-398 concentration increasing. The IC50 of 24 hours was 300 micromol/L. The release of PGE2 was significantly reduced in HepG2 cells with the values of NS-398 being (0.70 +/- 0.02), (0.48 +/- 0.02), (0.29 +/- 0.01) and (0.18 +/- 0.01) respectively, as compared with control group (0.03 +/- 0.01). NS-398 could inhibit the activity and expression of COX-2, with higher concentration, it can significantly down-regulate the expression of COX-2 (t = 3.736, 1.623, 1.810, 2.587, P < 0.01). CONCLUSION: NS-398 might significantly inhibit the proliferation of HepG2 cells and induce apoptosis. The mechanisms were related with the accumulation of quiescent G1 phase and the inhibition of COX-2 activity.

[Inhibition of curcumin on histone deacetylase and expression promotion of P21 (WAF1/CIP1) in HepG2 cells].

OBJECTIVE: To investigate the effect of curcumin (Cur) on histone deacetylase (HDAC1) and P21(WAF1/CIP1), a cyclin dependent kinase inhibitor, in HepG2 cells for exploring the mechanism of Cur in anti-cancer. METHOD: The HDAC1, P21(WAF1/CIP1) proteins and P21(WAF1/CIP1) mRNA were extracted from human hepatoma cells treated with or without Cur of different concentrations at different time points. Western blot analysis was performed to determine the levels of HDAC1 and P21(WAF1/CIP1) proteins, respectively. RT-PCR was performed to detect the level of P21(WAF1/CIP1) mRNA. RESULT: The IC50 of concentration treated by Cur was 25 micromol x L(1) on HepG2 cell. The level of HDAC1 was obviously inhibited by Cur, and decreased at 4 hours at IC, and lasted for 48 h in a time-dependent manner. The inhibition of HDAC1 was significant at the Cur concentration of 12.5 micromol x L(-1) but there was no difference between 50 and 100 micromol x L(-1). The levels of P21(WAF1/CIP1) mRNA and protein were up-regulated by Cur in dose and time-dependent manner, and the change of mRNA and protein was detected at 8 hours and lasted for 48 hours. CONCLUSION: Cur can inhibit the level of HDAC1 and enhance the expression of P21(WAF1/CIP1) protein and mRNA, and the results suggest that inhibiting HDAC1 and increasing P21(WAF1/CIP1) may be one of the possible mechanisms of anti-cancer by Cur.

Effect of manganese on heat stress protein synthesis of new-born rats.

AIM: To study the effect of manganese (Mn) on heat stress protein 70 (HSP70) synthesis in the brain and liver of new-born rats whose mother-rats were exposed to Mn. METHODS: 32 female rats were randomly divided into four groups. One group was administrated with physiological saline only as control group, the other three groups were administrated with 7.5, 15 and 30 mg x kg(-1) manganese chloride (MnCl2) by intraperitioneal injection every two days for two weeks. After delivery, the mother-rats received MnCl2 unceasingly for a week with the same method. Then the contents of Mn Zn Cu and Fe in the livers of the new-born rats were determined by atomic absorption spectroscopy; The level of HSP70 in the brains and the livers of the new-born rats as detected by Western-dot-blotting, and the SOD activities were measured simultaneously. RESULTS: The contents of Mn in the livers of new-born rats of the experimental groups(respective 1.38+/-0.18, 2.73+/-0.65,3.44+/-0.89 microg x g(-1)) were significantly increased compared with the control group(0.88+/-0.18 microg x g(-1); P<0.01); The contents of Fe in the livers of new-born rats of 15 and 30 mg x kg(-1) experimental groups (426+/-125, 572+/-175 microg x g(-1), respectively) were significantly increased compared with the control group(286+/-42 microg x g(-1); P<0.05); the levels of Zn in the livers of the new-born rats of three experimental groups(254+/-49, 263+/-47, 213+/-28 microg x g(-1), respectively) were lower than those of the control group(335+/-50 microg x g(-1); respective P<0.05, P<0.01); and the levels of Cu showed no significant difference among the four groups(three experimental groups: 75+/-21, 68+/-241 and 78+/-18 microg x g(-1); control group: 83+/-9 microg x g(-1); P<0.05). There was a significant increase in the levels of HSP70 in the brains of new-born rats of the 30 mg x kg(-1) group (19.5 x 10(3)+/- 1.3 x 10(3)A;control group:14.3 x 10(3)+/-1.4 x 10(3)A; P<0.01) and the levels of HSP70 in the livers of new-born rats of three experimental groups(respective 19.6 x 10(3)+/- 3.9 x 10(3)A,18.5 x 10(3)+/-3.8 x 10(3)A, 22.4 x 10(3)+/-1.9 x 10(3)A) also increased than control group(13.3 x 10(3)+/-1.0 x 10(3)A;P<0.01), but the SOD activities showed no significant difference among brains of the four groups (experimental groups: 5.04+/-0.43, 4.83+/-0.48, 4.60+/-0.84 ku x g(-1); control group: 4.91+/-0.37 ku x g(-1); P<0.05). The SOD activities in the livers of 15 mg x kgP< group(5.41+/-0.44 ku x gP<) was lower than the control group(5.95+/-0.36 ku x gP<; P<0.05). CONCLUSION: While mother-rats were exposed to manganese, the metabolisms of Mn Zn and Fe of new-born rats in the livers were influenced and were situated in a stress status, thus HSP70 syntheses is induced in the brains and livers of new-born rats, but the mechanism of this effect in the developmental toxicity of Mn remains to be further studied.