Ten compounds were isolated and purified by column chromatography over silica gel, preparative TLC, and Sephadex LH-20 from the whole plant of Solanum lyratum. The structures were elucidated on the basis of physico-chemical properties and spectral data as 1beta-hydroxy-1 ,2-dihydro-alpha-santonin (1) , boscialin (2) , blumenol C (3), 3beta-hydroxy-5alpha, 6alpha-epoxy-7-megastigmen-9-one(4), dehydrovomifoliol(5) , blumenol A(6), (1'S,2R,5S, 10R) -2-(1', 2'-dihydroxy-l1'-methylethyl) -6,10-dimethylspiro[4,5] dec-6-en-8-one(7) , (1'R,2R,5S,10R)-2-( 1',2'-dihydroxy-l '-methylethyl) -6,1 l0-dimethylspiro[4,5]dec-6-en-8-one( 8) , 2-(1',2'-dihydroxy-1 '-methylethyl) -6,1 0-dimethyl-9-hydroxyspiro [4,5] dec-6-en-8-one (9) , and grasshopper ketone (10). Compounds 1-10 were isolated from this plant for the first time.
Drugs, Chinese Herbal ; chemistry ; Solanum ; chemistry ; Terpenes ; analysis ; isolation & purification

The main problems and solutions of mineral drugs in clinical use were studied based on the investigation on the quality of Haematitum and Fluoritum pieces in venalicium. The outward appearance and intrinsic quality of Haematitum and Fluoritum pieces in venalicium were studied by the requires which were in the first part of Chinese Pharmacopoeia published in 2010. The outward appearance and intrinsic quality of mineral drugs had large differences, the disqualification rate was 41.67% in 12 batches of Haematitum pieces and the disqualification rate was 53.85% in 13 batches of Fluoritum pieces. The crushing granularity of mineral drugs should be defined, the quality standards should be further improved. The drug adiministration ought to strengthen inspection and supervision in order to ensure the stability and reliability of the clinical efficacy.
Medicine, Chinese Traditional ; standards ; Minerals ; standards ; Quality Control

Objective To develop a rapid,sensitive,specific and accurate quantitative duplex real-time PCR assay for detection of Listeria monocytogenes and Shigella.Methods Two sets of specific primers and probes were selected according to Listeria monocytogencs hly gene and Shigella ipaH gene.The target hly and iPaH fragments were amplified by PCR,and used to construct recombinant pGEM-T-hly and pGEM-T-ipaH respectively.The two recombinant circular plasmid DNAs were linearized with EcoR I that did not cut within the target DNA fragment.The ten-fold dilutions of plasmid were subjected to the standard quantitation curve in duplex real-time PCR assay.Various genomic DNAs of Listeria innocua,Listeria weshimeri,Salmonella,Staphylococcus aureus,Bacillus subtilis,Escherichia coli and Proteus were used as negative controls to confirm the specificity of duplex real-time PCR assay.The assay was also used to detect Listeria monocytogenes and Shigella in artificially contaminated sterilized skim milk.Results The recombinant plasmids were constructed successfully,hly probe(rAM and TAMRA double labelled)and ipaH probe (HEX and TAMRA double labelled)were used to develop an optimized PCR successfuliv.Conclusion The selected primers and probes showed high specificity for these two target bacteria,the linear range of the assay was good(105-101 copies/μl,R2≥0.998)and sensitivity Was 10 copies/PCR.Following a DNA extraction method which combined EZ Spin Colum Genomic DNA Isolation Kit(BBI)/Phenol-chloroform,the sensitivity of assay Was 102CFU/ml for both Listeria monocytogenes and Shigella in artificially contaminated sterilized skim milk,which equivalents to 10 CFU/PCR.

Objective To provide a basis for preventive strategies to national drinking-water-borne endemic arsenicosis through mastering the implementing progress of preventive measures and observing the dynamic changes.Methods Surveillances were carried out according to the provisions and requirements of The Surveillance Project for National Drinking-Water-Borne Endemic Arsenicosis(Trial).Total of 11 provinces(autonomous regions) and Xinjiang Production and Construction Corps were selected as the surveillance provinces (autonomous regions).Endemic arsenicosis villages with exposed population over 100 persons were chosen as monitoring villages in each province,81 villages in 2010 and 89 villages in 2011 were selected.Potential endemic arsenicosis villages with exposed population over 100 persons were included; 26 villages in 2010 and 19 villages in 2011 were selected.The operation of water-improving projects was investigated,the arsenic content in water from resident house was tested in potential endemic arsenicosis villages and the prevalence of endemic arsenicosis based on the residents who lived in monitoring villages was surveyed:Results ①Total of 225 water-improving projects in 45 counties were monitored,1349 villages were covered and 72.66 million persons were benefited in 2010.Total of 233 waterimproving projects in 48 counties were monitored,1576 villages were covered and 84.61 million persons were benefited in 2011.②)Total of 107 villages with high level of water arsenic were investigated and 81 villages had improved the water quality in these villages in 2010.The water-improving projects running normally reached 90.12％(73/81),intermittent operation rate was 9.88％ (8/81) and without abandoned projects.The projects with qualified water reached 86.42％ (70/81).Total of 108 villages with high level of water arsenic were investigated and 89 villages with water improved in 2011.Normally operated projects reached 86.52％ (77/89),intermittent operation rate was 11.24％ (10/89)and abandoned projects was 2.25％ (2/89).The projects with qualified water arsenic level reached 82.02％(73/89).In addition,26 villages without water-improvement were investigated in 2010,and the families with high level of water arsenic reached 66.01％(371/562).Total of 19 villages were surveyed in 2011,and the families with high level of arsenic reached 54.99％(204/371).③Total of 23 964 persons were examined in villages with improved water in 2010,the detection rate of patients with endemic arsenicosis was 4.43％ (1061/23 964),3964 persons were examined in the villages without water-improvement and the detection rate was 5.98％(237/3964),two new cases were diagnosed.Total of 25 225 persons were examined in villages with waterimproved,the detection rate was 4.68％(1181/25 225),3145 persons were examined in the villages without waterimprovement,and the detection rate was 2.26％(71/3145) in 2011,none new case was detected.Conclusions It is not optimistic about the operating status and quality of water-improving projects.The prevalence in water-improved villages remains higher than that in water-unimproved villages.The long-term mechanism of surveillance should be established and perfected as soon as possible,and the management and maintenance of water-improving projects also should be strengthened.

Objective To observe the expressions of inducible nitric oxide synthase(iNOS) in the progress of rat subchronic fluorosis,and analyse the mechanism of nitric oxide(NO) free radical injnry in bone.Methods Male wistar rats were divided randomly by body weight into two groups.i.e.sodium fluoride group and control group.Sodium fluoride group was given drinking water with 150 mg/L sodium fluoride,and control group was given tap water only.The animals were bred for 24 weeks.Every four weeks some rats were killed.The contents of serum and bone fluoride were examined and analyzed.The levels of serum NO were determined by Griess Reagent.The expressions of iNOS mRNA and protein were analyzed by RT-PCR and immunohistochemistry.Results The serum NO contents significantly increased(t=9.36,P＜0.01) in NaF-treated rats after 8 weeks[(19.94±3.04)nmol/L],but significantly decreased(t=10.47,4.46,P＜0.01) after 20 weeks[(11.55±3.54)nmol/L]and 24 weeks[(20.83±2.49)nmol/L],compared with control group[(9.11±1.21,31.13±3.93,33.10±7.37)nmol/L].The expression of iNOS mRNA significantly increased(t=13.09,4.82,14.23,4.64,7.82,5.29,P＜0.01)in rats treated with sodium fluoride[(1.87±0.11),(1.87±0.78),(1.90±0.29),(1.93±0.67),(1.88±0.38),(1.84±0.03)],compared with control group[(0.41±0.25),(0.30±0.17),(0.18±0.06),(0.63±0.15),(0.66±0.04),(0.65±0.55)],and these proteins mainly appeared in hyperplasie zone and hypertrophic zone cells of epiphyseal plate,cartilages,articular cartilage cells,osteoblasts and ligament cells.Conclusions High dose fluoride might persistentlv induce the expressions of iNOS and catalyze synthesis of NO,then regulates osteoblast and osteoclast activitv and finally influences bone turnover.

Objective To observe the expressions of matrix metal proteinases-13(MMP-13) mRNA and tissue inhibitor of metal protease- 1 (TIMP- 1) mRNA and analyse the molecular mechanism of bone matrix degradation in the progress of rat subchronic fluorosis. Methods Male Wistar rats were randomly divided into two groups according to body weight, i.e. sodium floride group and control group. Rats in the sodium fluoride group were given drinking water containing 150 mg/L F-, and the animals in the control group were given tap water. The animals were bred for 24 weeks. Every 4 weeks some rats were killed. The change of obsteoclst was observed by transmission electron microscope. The expression levels of MMP-13 mRNA and TIMP-I mRNA were analyzed by RT-PCR. Results The number of lysesome and the synthesis of lysosoma enzyme in osteeclast were decreased. The expression of MMP-13 mRNA was significantly increased(t=2.29,2.41,3.07,2.52, 3.15,2.22, P<0.05) in rats treated with sodium fluoride (1.87±0.67,1.87±0.75,1.90±0.73,1.93±0.86,1.88±0.61,1.84±0.53), compared with control group(1.24±0.39, 1.19±0.27,1.07±0.22, I. 15 ~ 0.17, 1.17±0.18, 1.20±0.62). The expression of TIMP-1 mRNA was significantly increased (t=2.69,2.19,2.68,2.46,2.43,2.96, P<0.05) in rats treated with sodium fluoride(1.89±0.77,1.70±0.85,1.61±0.82,1.81±0.84,1.70±0.74, 2.06±0.96), compared with control group (1.07±0.39,0.87±0.49,0.71±0.48,0.99±0.43,0.95±0.46,0.89±0.57). Conclusion High dose fluoride might persistently induce the expressions of MMP-13 mRNA and TIMP-1 mRNA and may be involved in bone turnover.

Objective To investigate the effect of aluminum on hair loss induc ed by fluoride in fluorosis mice.Methods Sixty male C57BL mice were divided into four groups according to body mass:control group,fluoride (F) group (F-100 mg/L),aluminum(Al) group(Al3+ 270 mg/L) and F + Al group(F-100 mg/L + Al3+270 mg/L).Mice were killed 1 month and 3 months after the experiment,respectively.Bone F content was detected by ion-selective electrode method.The level of bone Al was measured through inductively coupled plasma emission spectrum.Dental fluorosis and hair loss of mice were evaluated by visual method.Results One month after the experiment,no dental fluorosis and hair loss was found in all four groups.The content of bone F was the highest in F group [(2401.649 + 86.835) mg/kg],and the lowest in A1 group [(427.006 + 11.878) mg/kg].The levels of bone F in F + Al group and control group were (1210.332 + 19.531)mg/kg and (538.001 + 33.337)mg/kg,respectively.The difference was statistically significant between any two groups (all P ＜ 0.05).Three month after the experiment,all mice of F treatment group had dental fluorosis and hair loss(10/10).Alopecia areas were found in the neck and back regions only.There was no hair loss in control group,Al group and F + Al group.No dental fluorosis was found in both control and Al groups.Only 2 mice were found with dental fluorosis in F + Al group.The levels of bone F in F group,F + Al group,control group and Al group were (4098.645 + 58.842),(1888.165 ± 12.187),(876.258 + 14.462) and (662.385 ± 8.966) mg/kg,respectively.The difference was statistically significant between any two groups (all P ＜ 0.05).Conclusions The hair loss is found in fluorosis mice.Hair loss of mice is closely associated with the level of F exposure.Al can prevent the occurrence of hair loss induced by F in mice through reducing the accumulation of F.

Objective To grasp the operation quality of the water-improving projects for lowing fluoride level and to grasp the prevalence trend of fluorosis in drinking-water-borne fluorosis affected areas in China.Methods According to theSurveillance Scheme of Drinking-Water-Borne Endemic Fluorosis(Trial),314 counties were selected as monitoring counties in 2010 and 2011,respectively.Ten water-improving projects were randomly selected in each monitoring county every year.For each project,the operating condition was investigated and the fluoride level of tap water was determined.For each village,the fluoride level in drinking water was determined and the prevalence of dental fluorosis of all pupils aged 8-12 were surveyed.Dental fluorosis was diagnosed according to Dean method.Results ①A total of 1381 water-improving projects were monitored in 2010,among which normal operating projects accounted for 93.19％ (1287/1381) and those with qualified fluoride level accounted for 76.32％ (1054/1398).A total of 1408 water-improving projects were monitored in 2011.Normal operating projects accounted for 94.74％(1334/1408) and those with qualified fluoride level accounted for 79.26％ (1116/1408).②Total 319 villages with water-improving projects were monitored in 2010,and the number in 2011 was 327.The qualification rate of water fluoride content for 2010 and 2011 were 73.35％ (234/319) and 73.70％ (241/327),respectively.Total 84 villages without water-improving projects were monitored in 2010,and the number in 2011 was 88.Villages with fluoride level ＞ 1.2-2.0 mg/L,＞ 2.0-4.0 mg/L,and ＞ 4.0 mg/L in 2010 accounted for 47.62％ (40/84),42.86％ (36/84),and 9.52％ (8/84),respectively.The proportion in 2011 was 52.27％(46/88),38.64％(34/88),and 9.09％(8/88),respectively.③The detectable rates of dental fluorosis of children aged 8-12 in 2010 and 2011 were 26.97％(4347/16 119) and 24.98％(4027/16 123),respectively.In the monitored villages,the water-improving projects were under normal operation and fluoride content in water was qualified.And the rates for villages with water-improving projects under abnormal operation or fluoride content in water unqualified in 2010 and 2011 were 44.09％ (1721/3903) and 41.86％ (2188/5227),respectively.While the rates for villages without water-improving projects in 2010 and 2011 were 46.08％ (2802/6081) and 45.51％ (2804/6161),respectively.Conclusions The national normal operation rates of water-improving projects for lowing fluoride level are between 93％ and 95％.The qualification rate of fluoride content in drinking water is lower than 80％.The operation condition and water quality of the water-improving projects in China need to be improved.The prevalence of children dental fluorosis of affected villages has dropped significantly after low fluoride water is provided continuously.

Objective To investigate the prevalence of endemic arsenicosis and the progress made of control measures in China in 2006 so as to provide basis for endemic arsenicosis control in China.Methods The surveillance was carried out according to"the National Survey Scheme of Endemic Arsenicosis".The surveillance was carried out according to the national survey scheme of endemic arsenicosis.14 drinking water type of and 3 coal-burning type of endemic areas were selected.The fulfillment and effects of control measures,and prevalence of arsenicosi were sarveyed.Arsenic contents in drinking water,corn,chilli,coal and urine were detected by silver Burveyed had improved water was 1.04%(3/288)0f tlle pmjects fail to work,86.67%(247/285)of water arsenic concentration was aualifted.All the 8746 stoves were improved in the survey spots of coal-burning area.The rate of using high arsenic coal in survey spots was 51.08%(355/695).The qualified rate of stove was 75.05%(358/477), Among 14 survey spots,water arsenic concentrations of 6 spots were within safety threshold≤0.05 mg/L,in a rate of 42.86%.Arsellic concentration in some ofthe coal samples from Shiyakou Village,Anlong County was over 100 mg/kg. All chilli arsenic contents(mean)of 3 survey spots outnumbered national standard(≤0.5 mg/kg)and corn arsenic in drinking water areas in 2006,some of the pmjects are running out.The population of survey spots is still exposed of coal store tends to decline.Food arsenic pollution is mitigated compared with last year,especially chili.

Objective To observe the influence of fluoride and aluminum on the expression of matrix metalloproteinase-13(MMP-13) in rat articular chondrocytes. Methods Original generation chondrocytes of rats was cultured and divided into fluoride group, aluminum group, fluoride plus aluminum group and control group. NaF and A1C13 at concentrations of 1 mmol/L and 2 mmol/L were administered to intoxicate the cells for 24, 48, 72 h respectively. Cells were extracted to undergo reverse transcription the polymerase chain reaction(RT-PCR) at different times to observe mRNA expression of MMP-13, and protein expression was detected by Western-blot. Results In 24 h, the content of MMP-13 mRNA in fluoride group(0.830±0.043), aluminum group(1.279±0.060) and fluoride plus aluminum group(0.983±0.028) was higher than that in the control group(0.707±0.026, P<0.05), and relative expression of MMP-13 mRNA in aluminum group was the highest. In 48 h, the content of MMP-13 mRNA in fluoride group (0.964±0.180), aluminum group (1.333±0.105) and fluoride plus aluminum group (0.915±0.137) was higher than that in the control group(0.660±0.055, P<0.05), and the relative expression in aluminum group was the highest. In 72 h, the content of MMP-13 mRNA in fluoride group(0.866±0.115), aluminum group(0.846±0.089) and fluoride plus aluminum group(0.967±0.196) had no statistical significance(P>0.05) compared with the control group(0.809±0.179). In 24 h, the content of MMP-13 protein in fluoride group(1.050±0.084), aluminum group(1.010±0.113) and fluoride plus aluminum group(0.977±0.202) had no statistical significance(P>0.05) compared with the control group(0.860±0.038). In 48 h, the content of MMP-13 protein in fluoride group(0.671±0.020), aluminum group(1.134±0.094) and fluoride plus aluminum group (0.923±0.087) was higher than that in the control group (0.647±0.025, P<0.05), but no significant difference being observed between groups (P>0.05). In 72 h, the content of MMP-13 protein in fluoride group(0.672±0.022), aluminum group(1.088±0.072) and fluoride plus aluminum group(0.772±0.030) was higher than that in the control group(0.577±0.026, P<0.05). It was the highest in the aluminum group, the intra-group difference had statistical significance(P<0.05). Conclusions Fluoride and aluminum damage chondrocytes to some extent, toxicity of aluminum itself is greater than fluoride and fluoride plus aluminum. Abnormal expression of MMP-13 can be observed in the chondrocyte damage process induced by fluoride and aluminum.