The preliminary metabolic profile of total diterpene acid (TDA) isolated from Pseudolarix kaempferi was investigated by using in vivo and in vitro tests. Pseudolaric acid C2 (PC2) was identified as the predominant metabolite in plasma, urine, bile and feces after both oral and intravenous administrations to rats using HPLC-UV and HPLC-ESI/MS(n), and demethoxydeacetoxypseudolaric acid B (DDPB), a metabolite proposed to be the glucoside of PC2 (PC2G), as well as pseudolaric acid C (PC), pseudolaric acid A (PA), pseudolaric acid A O-beta-D glucopyranoside (PAG), pseudolaric acid B O-beta-D glucopyranoside (PBG) and deacetylpseudolaric acid A (DPA) originated from TDA could also be detected. It was demonstrated by tests that the metabolism of TDA is independent of intestinal microflora, and neither of pepsin and trypsin is in charge of metabolism of TDA, TDA is also stable in both pH environments of gastric tract and intestinal tract. The metabolites of TDA in whole blood in vitro incubation were found to be PC2, DDPB and PC2G, which demonstrated that the metabolic reaction of TDA in vivo is mainly occurred in blood and contributed to be the hydrolysis of plasma esterase to ester bond, as well as the glucosylation reaction. These results clarified the metabolic pathway of TDA for the first time, which is of great significance to the in vivo active form and acting mechanism research of P. kaempferi.
Animals ; Chromatography, High Pressure Liquid ; Diterpenes ; metabolism ; Glucosides ; metabolism ; Hydrolysis ; Mass Spectrometry ; Metabolic Networks and Pathways ; Pinaceae ; chemistry ; Rats

OBJECTIVETo investigate the effect of air pollution on stroke mortality in Tianjin, China, and to provide basis for stroke control and prevention.

METHODSTotal data of mortality surveillance were collected by Tianjin Centers for Disease Control and Prevention. Meteorological data and atmospheric pollution data were from Tianjin Meteorological Bureau and Tianjin Environmental Monitoring Center, respectively. Generalized additive Poisson regression model was used in time-series analysis on the relationship between air pollution and stroke mortality in Tianjin. Single-pollutant analysis and multi-pollutant analysis were performed after adjustment for confounding factors such as meteorological factors, long-term trend of death, "days of the week" effect and population.

RESULTSThe crude death rates of stroke in Tianjin were from 136.67 in 2001 to 160.01/100000 in 2009, with an escalating trend (P = 0.000), while the standardized mortality ratios of stroke in Tianjin were from 138.36 to 99.14/100000, with a declining trend (P = 0.000). An increase of 10 µg/m³ in daily average concentrations of atmospheric SO₂, NO₂ and PM₁₀ led to 1.0105 (95%CI: 1.0060 ∼ 1.0153), 1.0197 (95%CI: 1.0149 ∼ 1.0246) and 1.0064 (95%CI: 1.0052 ∼ 1.0077), respectively, in relative risks of stroke mortality. SO₂ effect peaked after 1-day exposure, while NO₂ and PM₁₀ effects did within 1 day.

CONCLUSIONAir pollution in Tianjin may increase the risk of stroke mortality in the population and induce acute onset of stroke. It is necessary to carry out air pollution control and allocate health resources rationally to reduce the hazard of stroke mortality.

Air Pollutants ; analysis ; Air Pollution ; analysis ; China ; epidemiology ; Environmental Monitoring ; Humans ; Models, Theoretical ; Particulate Matter ; analysis ; Poisson Distribution ; Stroke ; epidemiology ; mortality ; Survival Rate ; Time Factors

[Objects]To isolate and identify the pathogen of the large outbreak of dengue in Guangdong province in 2014. To understand the origin and the phylogenetic characteristics of the isolates ,and provide scientific foundation for the surveillance and prevention of dengue fever.[Methods]Collected the patient serum samples over all the Guangdong province during the 2014 outbreakperiod,isolated and identified the virus from these samples. Amplified complete E gene and complete genome with certain primers and sequenced all the products. Then the Phylogenetic ,Bayesian phylogeography and mutations analysis were carried.[Results]40 DENV-1 strains were isolated and identified. 40 complete E gene sequences and 6 complete genome sequences of DENV-1 were obtained. Phylogenetic analysis with E gene sequences revealed that the 40 isolates were classified into two genotypes including 16 genotypeⅠ(Asia)and 24 genotypeⅤ(America/Africa). 14 genotypeⅠisolates were clustered closest with isolates from Guangdong province(2013)and Sigapore(2013)which share the nucletide identities of 99.6% ~ 99.9%,other two genotypeⅠisolates were clustered with strains from Malaysia (2013) and both share the nucletide identities of 99.7%;24 genotypeⅤisolates were all classified in one clade with striains from Bangladesh(2009),China(2009)and Bhutan(2013)which share nucletide identities of 99.0%-99.9%. Further analysis with six complete genome sequences showed that five isolates were clustered closest with strains isolated from Guangdong province(2013)share the nucletide identities of 99.6%-99.8% while the sixth stains closest with strains isolated from Myanmar(2002)share the nucletide identities of 98.8%. The isolates have five amino acid mutations compared with strains epidemic in Guangdong province in 2013,three mutations(S88V,E203G,T275R)are in the EⅡdomain and one mutation (S305P)is in the EⅢdomain which associated with virulence.[Conclusions]During the outbreak in Guangdong province in 2014, DENV-1 is the predominant causative serotype,and there are at least two different kinds of genotypes of DENV-1 largely epidemiced in the whole province. Evolution analysis reveals the multiple origins of the isolates which may origin from Guangdong province , Sigapore,Malaysia,Myanmar so that we should enhance the study and surveillance of autochthonous and vectors in order to understand the epidemic way of dengue in Guangdong province. The isolates have had four mutations in the domain associated with virulence which remain further study to know their biological effects.

Using 16S rDNA clone library method, the bacteria composition of the Organic Matter-decomposing Inoculants A and B were investigated. The results indicated that: Sample A was clustered into 14 taxonomic operational units (OTUs),the dominant communities are Weissella confuse, Bacillus subtilis and Bacillus pumilus, which accounted for 28.6%,30.4% and 23.2%;Sample B was clustered into 43 OTUs,the dominant communities are Lactobacillus buchneri, Lactobacillus farciminis and Lactobacillus acetotolerans, which accounted for 18.03%,18.86% and 13.12%, respectively. The results had much difference from the samples' labels:there were not bacteria in the lable of sample A and only Bacillus pumilus was the same with the lable of sample B.This study showed that 16S rDNA clone library method has nicer application perspectives in analying the bacteria of microbial inoculant and its quality control.

Objective To quantitatively explore the effect of air pollution on coronary heart disease mortality in Tianjin.Methods Mortality data in 2001-2009 were from Tianjin mortality surveillance system operated by the Tianjin Centers for Disease Control and Prevention while data related to meteorology and air pollution were from the Tianjin Meteorological Bureau and Tianjin Environmental Monitoring Station,respectively.Generalized Additive Model (GAM) extended Poisson regression was used to examine the relationship between air pollution and mortality in Tianjin,under the controlling of time trends,weather,the day of week and population etc.Results The crude coronary heart disease mortality in Tianjin increased from year 2001 to 2009,from 105.12/100 000 to 167.03/100 000.The standardized mortality rate of 96.07/100 000 to 105.11/100 000.Air pollutants was more strongly associated with coronary heart disease mortality.By single GAM analysis,a 10 tg/m3 increase in SO2,PM10 and NO2,which accounted for 1.25％(95％CI:0.75％-1.75％),0.65％ (95％CI:0.51％-0.79％) and 1.04％ (95％CI:0.52％-1.55％) increased in daily mortality.By multiple GAM analysis,a 10 μ g/m3 increase in SO2 and PM10 which accounted for 0.86％ (95％ CI:0.60％-1.12％) and 0.40％ (95％ CI:0.06％-0.75％) increased the daily mortality.NO2 was not significantly associated with coronary heart disease mortality rates,but factors as low temperature,low wind speed and population size were (P＜0.05).Conclusion Results from this study showed that air pollution was a risk factor for coronary heart disease mortality in Tianjin.

OBJECTIVETo evaluate the different effect on the expression of Calcitonin gene related peptide (CGRP)and neuropeptide Y (NPY) between tissue engineered bone with vascular bundle graft in vivo and that with sensory nerve tract graft in vivo.

METHODThirty-six healthy New Zealand rabbits were divided into 3 groups randomly and equally: vascular bundle group (A), sensory nerve tract group (B), tissue-engineering group (C). Group A segmental bone defect of 1.5 cm long was made at the right femur in each animal. After plate fixation, the defects were implanted respectively with the engineered bone prepared in the above-mentioned 3 methods. At 3, 6 and 12 months post-operatively, the distribution of CGRP and NPY in the new bone were detected by immunohistochemistry and analyzed semi-quantitatively by image analysis software.

RESULTSCGRP and NPY immuno-histochemical results indicated their contents increased significantly in all 3 groups as time passed (P = 0.000). Compared with group B, the contents of CGRP and NPY in group A significantly increased at 3 months (P = 0.000), but there was no statistic difference between them at 6 or 12 months (P > 0.05). The expression of CGRP and NPY in both group A and B were significantly more than that in group C at 3, 6 or 12 months (P = 0.000).

CONCLUSIONImplantation of vascular bundle into tissue-engineered bone can significantly improve the CGRP and NPY contents at early 3 months comparing with Implantation of sensory tract into tissue-engineered bone, but the changes are not significant at 6 or 12 months post-operatively.

Animals ; Blood Vessels ; transplantation ; Bone Substitutes ; Calcitonin Gene-Related Peptide ; metabolism ; Disease Models, Animal ; Femur ; injuries ; Male ; Neuropeptide Y ; metabolism ; Peripheral Nerves ; transplantation ; Rabbits ; Random Allocation ; Tissue Engineering

BACKGROUNDThe synthesis of virus-like particles (VLPs) provides an important tool to determine the structural requirements for viral particle assembly and virus-host interactions. Our purpose was to express simultaneously all three structural proteins of hepatitis C virus (HCV) in insect cells to investigate the proteins assembly into VLPs and the immunogenicity of these particles.

METHODSHCV gene sequences encoding the structural proteins C, E1, and E2 were amplified with PCR, and recombinant baculoviruses were constructed using recombinant DNA techniques. The expression of HCV structural proteins in insect cells was analyzed by immunofluorescence and SDS-PAGE. The interaction of expressed structural proteins was investigated by immunoprecipitation and immunoblotting. The VLPs in the insect cells were visualized by electron microscopy (EM). VLPs were then purified by sucrose gradient centrifugation and used to immunize BALB/c mice. Antibodies against HCV were tested for in mouse serum samples by an ELISA assay.

RESULTSThe recombinant baculoviruses reBV/C and reBV/E1-E2 were constructed successfully. Insect cells co-infected with reBV/C and reBV/E1-E2 expressed HCV C, E1, and E2 proteins with the expected molecular weights of 20kD, 35kD, and 66kD, respectively. The results of immunoprecipitation and immunoblotting assays revealed the coimmunoprecipitation of C, E1, and E2 proteins, indicating association of the three structural proteins. Electron microscopy of insect cells co-infected with reBV/C and reBV/E1-E2 demonstrated spherical particles (40 to 60 nm in diameter) similar to the HCV virions from serum samples or hepatic tissue samples of HCV infected humans. The VLPs were partially purified. Antibodies to HCV were detectable in the serum of mice immunized with VLPs.

CONCLUSIONHCV structural proteins simultaneously expressed in insect cells can interact with each other and assemble into HCV-like particles, which are shown to be immunogenic in mice.

Animals ; Fluorescent Antibody Technique ; Hepacivirus ; immunology ; physiology ; Immunization ; Immunoprecipitation ; Mice ; Mice, Inbred BALB C ; Microscopy, Electron ; Spodoptera ; Viral Structural Proteins ; immunology ; physiology ; Virion ; physiology ; Virus Assembly

OBJECTIVE: To discuss the cause of disease, treatment and therapeutic effect in patients with rhegmatogenous retina detachment (RRD) combined with non-secondary glaucoma. METHODS: Clinical data of 28 patients with RRD combined with primary or congenital glaucoma were retrospectively analyzed. RESULTS: Twenty-five out of the 28 patients succeeded with one operation (89.3%). The intraocular pressure of post-operation:on the 1st day was 10 approximately 46 (28.1+/-6.5) mmHg, on the 7th day was (18.9+/-7.2) mmHg, and on the last re-examination day was (17.6+/-6.2) mmHg. Anti-glaucoma operation was performed in 10 patients after the retinal operation. Chroidal hemorrhage was found in 2 patients and 2 chroidal exudations were found after the retinal operation. CONCLUSION The proportion of primary open angle glaucoma is higher than that of primary angle closure glaucoma, and trauma or surgery before the retinal operation is an important cause in glaucoma patients with RRD. There is no obvious difference in the ratio of surgical success between non-secondary glaucoma with RRD and those RRD patients without glaucoma. Vitreotomy+ silicon oil injection or drainage of subretinal fluid+air injection+cryocoagulation+explants is recommended. Chroid is easily involved. It is important to control the intraocular pressure during and after the surgery. The final visual acuity is rather poor, which may be related to the glaucoma and intraocular pressure.
Adolescent ; Adult ; Aged ; Child ; Female ; Glaucoma ; complications ; surgery ; Humans ; Male ; Middle Aged ; Retinal Detachment ; complications ; surgery ; Retrospective Studies ; Visual Acuity ; Vitrectomy

To develop a model for forecasting the mortality of stroke in Tianjin,China.The time series of stroke mortality from 1999 Jan.to 2006 Dec.in Tianjin city were subjected.Circle distribution analysis was used to verify the trend of time concentration.Multiple seasonal autoregressive integrated moving average model [ARIMA (p,d,q) (P,D,Q)s],based on model identification,estimation and verification of parameter,and analysis of the fitting of model,was established.Most of the deaths from stroke occurred in January and had a cycle of 12 months.An AR/MA model (0,1,0)×(0,1,1)12 was established(1-B)(1-B12) lnxt=0.001+(1-0.537 B12)εt.Conclusion: ARIMA & Circle Distribution analysis is an important tool for stroke mortality analysis.Potentially it has a high practical value on the surveillance,forecasting and prevention of stroke mortality.

OBJECTIVETo observe the blood enriching function of Siwu Tang and its effect on Epo and G-CSF gene expressions in bone marrow of blood deficiency mice, and thus provide the basis for understanding the molecular mechanism of blood enriching function of Siwu Tang.

METHODThe animal model of blood deficiency were established in the mice by using 3.5 Gy60Co gamma-ray. Peripheral blood cells were analyzed and CFU-GM, BFU-E, CFU-E and CFU-mix were counted in bone marrow colony cultured. Both Epo and G-CSF gene expressions in bone marrow were measured with RT-PCR.

RESULTSiwu Tang significantly increased the number of peripheral blood cells and the amount of CFU-GM, BFU-E, CFU-E and CFU-mix in bone marrow and enhanced Epo and G-CSF gene expression in bone marrow in the mice with blood deficiency.

CONCLUSIONThe promotion of Epo and G-CSF gene expressions in bone marrow may be one of the mechanisms underlying the blood enriching function of Siwu Tang decoction.

Animals ; Bone Marrow ; metabolism ; Bone Marrow Cells ; cytology ; Cell Count ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Erythroid Precursor Cells ; cytology ; Erythropoietin ; biosynthesis ; genetics ; Female ; Granulocyte Colony-Stimulating Factor ; biosynthesis ; genetics ; Leukocyte Count ; Medicine, Chinese Traditional ; Mice ; Mice, Inbred C57BL ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Whole-Body Irradiation