Objective To discover the mutation of ADAR gene in a pedigree with dyschromatosis symmetrical hereditaria(DSH). Methods We investigated this family and collected blood samples of the individuals in this family. Mutation screening was carried out by PCR and direct sequencing. The allele specific primer was designed for the mutation point, and allele-specific PCR was carried out on the patients, normal family members and 40 normal individuals. Results A single nucleotide deletion (c.1642 delC) was identified in exon3 of ADAR gene in the patients of this family. This mutation was not detected in the normal family members and in any of the control individuals. Conclusion This single nucleotide deletion was responsible for the disease in the family.

AIM: To explore the effect of Buddleia flavonoids drug-containing plasma and androgen receptor(AR) blocker on the expression of STAT1 phosphoprotein.METHODS: In vitro lacrimal gland epithelial cells were cultivated with H2O2 to establish the dry eye apoptosis state. Blank plasma group, Buddleia officinalis plasma total flavonoids interfere with drug-containing group, and the intervention group of testosterone propionate were set. The expressions of STAT1 phosphoprotein of each group were observed by Western blot and AR blocker flutamide was used to explore the intended androgen effect of Buddleia flavonoids.RESULTS: After the intervention of drug-containing plasma, the expression of STAT1 Phosphoprotein in Buddleja officinalis drug-containing plasma intervention group(0.353±0.494) and testosterone propionate intervention group(0.502±0.036) were enhanced and the differences between the two groups were significant (P<0.01). After using the AR blocker in all groups, the expression of STAT1 phosphoprotein in each group (0.268±0.061,0.283±0.106,0.213±0.071) had no difference.CONCLUSION: Buddleja officinalis drug-containing plasma total flavonoids can promote the expression of STAT1 phosphorylation.

Objectives To screen the differentially expressed proteins in serum of population chronically exposed to arsenic in drinking water,thus to provide candidate protein biomarkers for arsenic exposure and arsenicosis.Methods Subjects were selected from the drinking water type of endemic arsenicosis areas in Shanxi province,China.Demographic characteristics,history of arsenic exposure,cigarette smoking,alcohol drinking,health and other information were collected using questionnaire.The subjects were divided into low-arsenic group (with arsenic in drinking water ＜ 10 μg/L),medium-arsenic group( 10 - 50 μg/L),high-arsenic group( ＞ 50 μg/L),and arsenicosis group(the drinking water with arsenic ＞ 50 μg/L was replaced by low arsenic water ＜ 10 μg/L).The number of cases in each group was 30.The arsenicosis patients were diagnosed according to “Standard of Diagnosis for Endemic Arsenism” (WS/T 211-2001 ).With the principle of informed consent,blood samples were collected.Differentially expressed serum proteins of different arsenic exposure groups and arsenicosis group were screened by two-dimensional differential gel electrophoresis(2-D DIGE),and further identified by mass spectrometry (MS).Results An average of (1299 ± 167) protein spots were identified in 6 gel images and 688 protein spots were discovered repeatedly in at least 5 gels.There were 33 protein spots differentially expressed among low-,medium- and high-arsenic groups P ＜ 0.01).Fifty four protein spots were significantly different among low-,medium-,high-arsenic exposure groups and arsenicosis group(P ＜ 0.01 ).Twenty five protein spots were selected for MS analysis,and13 protein spots were identified.Compared with low-arsenic group,the expressions of apolipoprotein A-Ⅳ,retinol binding protein,and estrogen receptor hypothalamic isoform in medium- and higharsenic exposure groups were down regulated,and the expressions of component 4A and 4B were up regulated.Compared with low-,medium- and high-arsenic groups,the expressions of beta-2-glycoprotein Ⅰ,Keratin 1,hemopexin,complement C1r subcomponent,and ficolin-3 in arsenicosis group were down regulated,and the expressions of pigment epithelial-differentiating factor,alpha-1-microglobulin and carboxypeptidase N catalytic chain were up regulated.Conclusions Chronic arsenic exposure can significantly change population's serum protein expression.Differentially expressed proteins in arsenicosis patients will not decline with the decline of arsenic in a short term.Whether or not the differentially expressed proteins identified in this study can be used as biomarkers for arsenic exposure and arsenicosis needs to be further verified.

Objective To develop a rapid,reliable and convenient approach for diagnosing the homozygous deletion of SMN1 gene.Methods SMN1 gene was amplified specifically with double allele-specific PCR(AS-PCR).Meanwhile,one inrelevant gene was amplified as internal control by PAGE and agarose gel electrophoresis analysis to determine whether the sick children were with homozygous deletion of SMN1 genes.Results The homozygous deletion of exon7 in SMN1 gene was identified by agarose gel electrophoresis or PAGE accurately.Conclusion Compared to PCR-RFLP and DHPLC used in the past,this approach can diagnose homozygous deletion of SMA much more accurate,easier and more convenient without completed following analyses.

Objective To explore the proliferation characteristics of primary small intestinal epithelial cells of tree shrews and the characteristics of human rotavirus(RV) G1P[8] infection to these cells,and establish a model of tree shrew primary small intestinal epithelial cells infected with human rotavirus G1P[8].Methods The primary small intestinal epithelial cells were obtained by collagenase Ⅺ and dispase I digestion from tree shrew.After purification and identification,the obtained primary small intestinal epithelial cells were infected with RV.Then,culture supernatants of infected cells were collected every 12 hours after infection.Viral titer and viral load were subsequently determined.Western blot and indirect immunofluorescence observation were used to detect the expression of RV protein VP6 in the primary cells.The infectivity of RV to the tree shrew primary cells was finally evaluated.Results After purification and identification of primary epithelial cells from the tree shrew,high purity above 90% primary tree shrew small intestinal epithelial cells was obtained.These primary small intestinal epithelial cells could be infected with RV virus by comparing the virus infectivity to primary renal cells,HCT116 cells and MA104 cells.The virus titer reached to 2.0×105TCID 50/mL at 72 h after infection.Using Western blot and indirect immunofluorescence observation,the specific viral protein of VP6 was determined to be expressed in the tree shrew primary small intestinal epithelial cells,and were located in the cytoplasm from days 1 to 5.Conclusions The separation,purification and cultivation methods of tree shrew primary small intestinal epithelial cells are successful,and the tree shrew model of RV-infected the tree shrew primary small intestinal epithelial cells is successfully established.

Breeding is not only an important area of medicinal plants research but also the foundation for the superior varieties acquirement of medicinal plants. The rise of modern biotechnology provides good opportunities and new means for medicinal plants breeding research in China. Biotechnology shows its technical advantages and new development prospects in breeding of new medicinal plants varieties with high and stable yield, good quality, as well as stress-resistance. In this paper, we describe recent advances, problems, and development prospects about the application of modern biotechnology in medicinal plants breeding research in China.
Biotechnology ; methods ; Breeding ; China ; Plants, Medicinal ; genetics ; Research ; Tissue Culture Techniques

Objective To assess the laparoscopic technique combined with open surgical technique in pyeloplasty.Methods Overall,45 patients with ureteropelvic junction obstruction underwent laparo- scopic dissection of the renal pelvis and upper ureter transperitoneally,and pyeloplasty was performed through a expanded trocar-incision(extension of 1-2 cm)as open surgery was performed.Results The opera- tion was successful in all 45 patients.The mean operative time was 58 min(range,40-85 min),and the mean blood loss was 22 ml(range,15-30 ml).No complication was observed during and after operation. Follow-up for 3-36 months was available in 34 patients.Intravenous urography(IVU)showed no obstruc- tion of the anastomotic stoma,and B-ultrasound indicated relief of hydronephrosis.Conclusions Laparo- scopic approach combined with open surgery in pyeloplasty is an effective way to treat ureteropelvic junction obstruction.This technique can simplify the operative manipulation and shorten the operative time without more trauma to the patients.It is worth general application in clinical practice.

ObjectiveTo observe the changes of fluoride content in serum,bone and urine after rats were exposed to single fluoride,single aluminum or fluoride combined with aluminum and to investigate the effects of different doses of aluminum on fluoride accumulation and excretion in rats.Methods Male Wistar rats were randomly divided into 9 groups based on 3 × 3 factorial design.Different doses of fluoride(NaF,0,50,200 mg/L)and(or) aluminum(AlCl3,0,100,200 mg/L) were administered to rats in each group by drinking water.The rats took food and water ad libitum during the experimental period.After feeding for 18 weeks,rats with obvious dental fluorosis were determined as successful establishment of animal model.The fluoride content in the serum,bones and urine were measured.Results Fluoride affected the fluoride content in serum,bones and urine(F=166.74,577.81,160.96,all P ＜ 0.01 ).The interaction of fluoride and aluminum on serum,bone and urinary fluoride were statistically significant (F =7.95,5.13,6.94,all P ＜ 0.01 ).When the fluoride level was 50 mg/L,the serum fluoride contents were [ (0.08 ± 0.03) and (0.08 ± 0.02) mg/L] in the aluminum levels of 0 and 100 mg/L groups,which was higher than that of the aluminum level of 200 mg/L group[ (0.04 ± 0.01)mg/L,F=7.14,5.78.all P＜ 0.05].The bone fluoride content in the 0 mg/L aluminum level group[ (1996.53 ± 383.73) mg/kg] was higher than that of the 100 and 200 mg/L groups[(1252.51 ± 189.08),( 1160.63 ± 129.63) mg/kg,F=20.54,24.56,all P ＜ 0.01 ].When the fluoride level was 200 mg/L,the bone fluoride contents were decreased with the increasing doses of aluminum[ (4668.70 ± 887.67),(3920.30 ± 528.31 ),(3297.64 ± 396.04) mg/kg].Between any two groups,the differences were statistically significant (F =15.59,52.31,14.38,all P ＜ 0.01 ).When the fluoride level was 50 mg/L,the urinary fluoride content in the 0 mg/L aluminum level group[ (34.054 ± 9.30)mg/L] was higher than that of the 100,200 mg/L groups[( 14.81 ± 6.32),(14.67 ± 3.42) mg/L,F =25.30,24.32,all P ＜ 0.01 ].When the fluoride level was 200 mg/L,the urinary fluoride contents in the 0,100 mg/L aluminum level groups[ (57.14 ± 21.38),(51.75 ± 8.39)mg/L] were higher than that of the 200 mg/L group[(34.839 ± 9.30) mg/L,F=30.04,20.31,all P ＜ 0.01 ].ConclusionsAluminum is an antagonist of fluoride.The antagonism could be enhanced as the dose of aluminum increased.In this study,aluminum could effectively counteract the absorption of fluoride in rat model when the ratio of fluoride to aluminum is 1 ∶ 2.

Objective To investigate the occurrence of new cretinism cases and the prevalence of endemic goiter, and the reason of lower coverage rate of iodized salt in the iodine deficiency disorders(IDD) high-risk areas of China, so as to put forward target prevention measures for these areas. Methods A hundred and one counties from 11 provinces(autonomous regions, municipality), such as Tibet, Qinghai, Xinjiang, Gansu, Ningxia, Sichuan, Hainan, Chongqing, Yunnan, Guangxi, Inner Mongolia, were chosen into the survey by simple random sampling. In the counties of high risk, typical sampling principle was used. In the selected townships, searching for new cretinism cases were carried out in the children under 10 years old, the thyroid volume of children aged 8-10 years old were determined by B-ultrasonography methods and their urinary iodine (UI) were determined by As3-Ce<'4+> catalytic spectrophotometry, the intelligence quotient(IQ) values of children aged 8-10 years old were measured by the combined Raven Test in China. In the household survey, the housewives were asked to fill in the questionnaire, the iodized salt coverage rates and the UI levels of child-bearing age women were investigated, the salt iodine content was determined using self-quantitative kit. Epi Info software was used to analyze the determination results. Results In the 101 high-risk counties, 249 were diagnosed as new cretinism cases from 4122 suspected cases searched. The goiter rate of children aged 8-10 years old by B-ultrasound was 8.28% (4434/53 541), 44 counties had goiter rates in the range of 5%-20%, 5 counties had goiter rates in the range of 20%-30%, and 3 counties had goiter rates of 30%. The mean IQ of children was 85.44, and the percentage of IQ value less than 70 was 16.52%(8713/52 745). The median urinary iodine(MUI) of children was 154.69 μg/L, the percentage of UI less than 50 μg/L was 17.26% (9069/52 558). Twenty-five counties had a MUI of children less than 100 μg/L. The MUI of housewives was 107.14 μg/L, the percentage of UI less than 50 μg/L was 27.50% (3722/13 534). MUI of housewives in 46 counties were less than 100.0 μg/L. The coverage rate of iodized salt at household level was 77.85%(13 150/16 891). The coverage rate of iodized salt was 52.80%(1585/3002), 44.72% (631/1411) and 72.82% (1850/2506) in Tibet, Hainan and Qinghai, respectively. More than 10% residents of Tibet, Sichuan, Hainan, Gansu and Qinghai complained that iodized salt was not convenient to buy. There were 71.39%(7652/10 719) of observed people ate crude salt. The average price of crude salt price(0.30-1.20 Yuan/kg) was lower than iodized salt(1.20-3.00 Yuan/kg). Conclusions In these IDD high-risk areas, the risk of endemic goiter and cretinism prevalence is threatening. The IDD monitoring should be carried out successively in these high-risk areas. The prevention measures, increasing iodized salt coverage rate and establishing the sustainable mechanism for eliminating IDD should be strengthened. Emergent iodine fortification measure for high risk region people should be implemented as soon as possible, a long term effective mechanism of eliminating IDD should be established.

Autophagy, an evolutionarily conserved process by which components of the cell are degraded in lysosomes, may facilitate survival of cancer cells under stress conditions. 8-Azaguanine (8-AG), an inhibitor of purine nucleotide biosynthesis, shows antineoplastic activity in multiple tumor cells. However, chemoresistance has restricted its development as an anticancer agent, and the mechanism of 8-AG resistance is not fully understood. We report here that 8-AG induces a protective autophagy to eliminate its cytotoxicity, and inhibition of autophagy increases cellular sensitivity of cancer cells to 8-AG treatment. Using HepG2 or SMMC-7721 hepatic cancer cell lines, we found that 8-AG inhibited cell viability and induced intrinsic apoptosis, accompanied by the up-regulation of the pro-apoptotic protein BimS, one of Bim (also known as BCL-2-like protein 11, BCL2L11) isoforms. Furthermore, 8-AG treatment enhanced the autophagy flux by promoting the dephosphorylation and activation of Unc-51-like autophagy activating kinase 1 (ULK1) via Akt/mTORC1 (mammalian target of rapamycin complex 1) signaling inhibition. Depletion of autophagy-related gene 7 (ATG7) markedly enhanced the level of BimS, and promoted cell death in response to 8-AG. 8-AG in combination with autophagy inhibitor chloroquine (CQ) or bafilomycin A1 (Baf A1) promoted the 8-AG-induced apoptosis in hepatic cancer cells. Altogether, these findings suggest that autophagy promotes chemoresistance of cancer cells for 8-AG, and blocking autophagy increases cellular sensitivity of cancer cells to 8-AG treatment.