Acupuncture Points ; Adult ; Drugs, Chinese Herbal ; administration & dosage ; Ear ; anatomy & histology ; Female ; Humans ; Melanosis ; drug therapy ; Middle Aged ; Young Adult

Traditional recombination technology of bacteria chromosome and its limitation were introduced. The definition of Red recombination technology is put forward: a method of homologous recombination between foreign linear DNA and the target gene in chromosomes mediated by ? phage Red system. The linear DNA referred here is general PCR product or oligonucleotide, which has a 36~50bp homologous sequence with the target gene in chromosome at both flanking. Red recombination technology leaves out the in vitro DNA restriction enzyme digestion and link process, which makes the knockout and alternation of target gene in bacteria chromosome relatively easier, and becomes an effective method to exploring genes and constructing new strains gradually. The gene inactivation and alternation method aiming at bacteria chromosome applied to Red recombination system was summarized by the structure element, action mechanism, and strategy of recombination, advantage and developing prospect. The Red system includes three genes: bet (aka?), exo and gam (aka ?). Exo is a 5′→3′ exonuclease, which degrades the 5′ ends of linear DNA molecules. Bet is a single-stranded DNA binding protein that binds to the single stranded 3′ ends generated by Exo and promotes annealing to complementary DNA. Gam binds to the host RecBCD complex and inhibits its exonuclease activity. Red recombination system may be constructed in such plasmids as pKD20 and pKD46 or in chromosome of bacteria. Most bacteria are not readily transformable with linear DNA because of the presence of intracellular exonucleases that degrade linear DNA. But when bacteria cells are transformed with pKD20 or pKD46 plasmid, or integrated with a detective ? prophage, Red recombination enzymes may be expressed in host cells, which make linear DNA with 36~50bp extensions that are homologous to both flanking of target genes transform E.coli readily and knock-out or alternate target gene. The Red recombination method is not only useful in chromosomal gene inactivation in E.coli, but also in other bacteria or virus, such as Salmonella, Shigella flexneri and virus HaSNPV. With the proceeding research, Red system will be applied for more and more purposes, and contribute a lot for gene improvement and gene function investigation in the coming Postgenome Era.

Objective To explore the plague epidemical trend of nearly a 10 years data in Qinghai province to provide basis for making the prevention and control measures. Method The regional distribution and time distribution of animal and human plague, monitoring and plague foci of survey data in Qinghai from 2001 to 2010 were analyzed with Excel software 2003. Results In Qinghai province, a total of 167 strains of Yersinia pestis were isolated from infected animals and insects in 10 years. Yersinia pestis was mainly distributed in Wulan,Delinha, Geermu, and Tianjun, along the Qinghai-Xizang railway. Human plague was occurred every year from 2001 to 2010 except 2002, 2007, 2008, and 2010. In the 10 years, there were 37 plague cases and 16 of these cases died, the mortality was 43.24％. The plague cases were mainly distributed in Nangqian, Qumalai, Chenduo,Zhiduo, Xinghai, Tongde, Tianjun, Wulan and Qilian. And these cases were found mostly in the period from May to October, especially in the period from August to October. Major clinical type of the plague cases was lung-type (62.16％,23/37). Conclusions The plague epidemic situation in Qinghai province is still severe, animal plague occurred year after year, and human plague outbreaks occasionally. Monitoring and early warning in the key areas should be strengthened, and the comprehensive measures of plague prevention and control should be carried out to reduce the incidence and prevalence of plague.

Objective To determine effects of T-2 toxin and selenium on expression of aggrecanase in human chondrocyte.Methods Human chondrocytes were treated with T-2 toxin(0,1,10,20 μg/L),and/or sodium selenite(final concentration of selenium 0,0.1 mg/L) for 5 days.Aggrecan expression was determined by Western blotting,aggrecanase-1 and aggrecanase-2 mRNA levels were measured by RT-PCR.ResultsSelenium and T-2 toxin had effects on both aggrecan protein levels and its aggrecanases(include aggrecanase-1 and aggrecanase-2 ) mRNA levels(F =0.294,27.71 for aggrecan,F =107.45,362.25 for aggrecanase-l,F =34.68,22.26 for aggrecanase-2,respectively,all P ＜ 0.05),and there was interaction between selenium and T-2 toxin on aggrecan,aggrecanase-1 and aggrecanase-2 expression(F =79.99,230.76,388.33,all P ＜ 0.05).Furthermore,selenium presented significant antagonism to T-2 toxin on aggrecan,aggrecanase-1 and aggrecanase-2 expression.Aggrecan expression levels(0.278 ± 0.015,0.235 ± 0.029,0.195 ± 0.028,0.399 ± 0.028,0.299 ± 0.020,0.263 ±0.019) induced by both 1,10,20 μg/L T-2 toxin and 0,0.1 mg/L selenium were significantly decreased than the levels(0.472 ± 0.0358,0.197 ± 0.018,all P ＜ 0.05) in control group(0 mg/L toxin).Selenium partially blocked the effects induced by 1,10,and 20 μg/L T-2 toxin(all P＜ 0.05).One,10,20 μg/L T-2 toxin and 0,0.1 mg/L selenium increased both aggrecanase-1 mRNA levels(0.535 ± 0.033,1.071 ± 0.043,1.454 ± 0.058,1.057 ±0.048,0.555 ± 0.036,0.902 ± 0.045) and aggrecanase-2 mRNA levels(0.596 ± 0.038,0.656 ± 0.033,0.949 ±0.049,0.600 ± 0.040,0.453 ± 0.031,0.164 ± 0.011),when compared with control(0.481 ± 0.023,0.346 ±0.020 for aggrecanase-1 and 0.387 ± 0.020,1.021 ± 0.046 for aggrecanase-2,respectively,all P ＜ 0.05).Selenium partially blocked 10,20 μg/L T-2 toxins induced upregulation of aggrecanase-1 (all P ＜ 0.05) and aggrecanase-2 (all P ＜ 0.05 ).Conclusions These data suggest a possible molecular mechanism that T-2 toxin could induce cartilage matrix degradation through the upregulation of aggrecanases expression and enzyme activities.Trace element selenium has some protective effect on cartilage proteoglycan degradation induced by T-2 toxins.

Objective Occult hepatitis B infection of voluntary blood donors has been plagued in the serum screening.Determined the OBI through the highly sensitive detection methods Nest-PCR among the blood donors,and then learned occult HBV infection and analysed the genotypes of this area.Methods 10 080 serums of donors were determined respectively by the imported Abbott HBsAg kit and Beijing Wantai anti-HBc and anti-HBs reagents,obtained the gene and detected DNA sequences by the high sensitive Nest-PCR method.Results Among 10 080 cases of unpaid blood donors,108 cases were detected HBsAg positively by Abbott sensitivity kit ( positive rate of 1.07％ ),767 cases were anti-HBc single - positive ( positive rate of 7.67％ ).25 patients screened blood donors who tested negative for serum HBsAg and positive for HBV DNA in the 10080 cases.Occult HBV infection incidence rate was 0.25％.12 cases were HBV genotype C (48％),13 cases were genotype B (52％),and no other genotypes.Genotype B has no statistically significant difference to genotype C ( P ＞ 0.05 ).Sequence analysis showed that 5 patients in the HBsAg epitope "a" (aa124 - aa147 ) have mutation (20％).Conclusion The high proportion of occult hepatitis B infected among voluntary blood donors in our country.Also genotype and mutation was differences in different regions.

Objective To assess if the modulating effect of platelet-derived growth factor(PDGF)BB on p21~(WAF1) was mediated by upregulating transforming growth factor(TGF)-β_1 expression in vascular smooth muscle cells(VSMC).Methods TGF-β_1 mRNA and protein expressions were measured by reverse transeription-PCR and ELISA,the protein expressions of p21~(WAF1) and the downstream TGF-βsignalling including TGF-β type I receptor(ALK-5 in VSMC),Smurf2,pSmad2/3,Smad4,Smad7 were detected by Western blot.Results PDGF-BB significantly upregulated the expressions of TGF-β_1 at mRNA(0.79-fold)and protein(1.98-fold)levels in VSMC,significantly inhibited the expression of p21WAF1(-67±12)%,and enhanced the expressions of ALK-5,pSmad2/3,Smad4,Smurf2 protein by 1.21-fold.0.95-fold,0.69.fold and 2.55-fold respectively,inhibited Smad7 expression(-65±9)%,these alterations were partially restored by anti-TGF-β_1 neutralizing antibody.Conclusions These findings suggested that PDGF-BB inhibited p21~(WAF1) expression in VSMC partially through upregulating TGF-β_1 expression via PDGFBB and TGF-β signalling pathways.

OBJECTIVETo study effects of alternative transcripts of ING1 transfection on human cancer cell lines.

METHODSp47/ING1A and p33/ING1B expression vehicles were constructed and introduced into a human breast cancer cell line MCF-7 and a human lung cancer cell line PAa, both expressing wild-type p53 protein. Growth characteristics of the transfectants and potentially related genes were analyzed.

RESULTSThe levels of p47/ING1A and p33/ING1B protein elevated respectively in tumor cells of MCF-7 and PAa after transfected with p47/ING1A and p33/ING1B, and the latter was much higher than that of the former. Ectopic overexpression of p33/ING1B effectively blocked tumor cell growth and arrested cells in the G(0) approximately G(1) phase of the cell cycle (P < 0.01), while p47/ING1A gave no effect on cell growth or cell cycle. Tumor cells overexpressing p33/ING1B contained more p21(WAF1) protein than that of the control cells, with undisturbed p53 protein level.

CONCLUSIONSExpression of two different transcripts of ING1 may have different effects on tumor cell growth. p33/ING1B may cooperate with p53 in stimulating expression of p21(WAF1) gene, thus to arrest cell cycle and to inhibit tumor cell growth. p33/ING1B may be considered to be a candidate as a partner of p53 in gene therapy.

Adenocarcinoma ; genetics ; metabolism ; pathology ; Alternative Splicing ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Cycle ; Cell Cycle Proteins ; Cell Division ; Cell Line, Tumor ; Cyclin-Dependent Kinase Inhibitor p21 ; Cyclins ; biosynthesis ; DNA-Binding Proteins ; Genes, Tumor Suppressor ; Humans ; Inhibitor of Growth Protein 1 ; Intracellular Signaling Peptides and Proteins ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Nuclear Proteins ; Protein Biosynthesis ; Proteins ; genetics ; Transfection ; Tumor Suppressor Protein p53 ; biosynthesis ; Tumor Suppressor Proteins

OBJECTIVETo observe the effect on intervention of sub-health with pestle needle (Chuzhen).

METHODRandomized controlled trail was adopted for this research. One hundred and fifty-three cases were randomly divided into two groups of a Chuzhen group (79 cases) and a massage group (74 cases). Acupoint of Bazhen (Baihui Bazhen, Shendao Bazhen, Zhiyang Bazhen, Mingmen Bazhen, Yaoyangguan Bazhen), Hechelu on the head, the neck and the lumbar area were adopted in Chuzhen group. While regular whole-body massage was applied in the massage group. The human sub-health score, the cornell medical index (CMI) and thermal texture maps system (TTM) technology of the two groups before and after the intervention were observed.

RESULTS1) After treatment, sub-health condition score, the CMI score, the M-R score and the TTM index were all increased in both groups (all P<0.01) 2) Comparison of D-value of the two groups before and after the intervention: the level of the sub-health score, the total score of CMI, and the index of sleep, pressure, Governor Vessel, Hukou (first web), blood lipid, viscosity of blood, microcirculation of TTM index of the Chuzhen group changed more obvious (all P<0.01), but there was no statistic significances in the M-R score and blood sugar of the TTM (both P>0.05). 3) The sub-health condition score in Chuzhen group was higher than that in the massage group (P<0.01).

CONCLUSIONChuzhen therapy has definite effect on intervention of sub-health, which is better than regular general massage.

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Cornell Medical Index ; Female ; Humans ; Male ; Massage ; Middle Aged ; Young Adult

OBJECTIVEBy using Benchmark Dose (BMD) approach to explore the relations among drinking water fluoride, urine fluoride, serum fluoride and dental fluorosis; and to evaluate the significance of urine fluoride and serum fluoride in control and prevention of endemic fluorosis.

METHODS512 children (290 in Xinhuai Village, 222 in Wamiao Village) aged 8-13 years were recruited in the study. Epidemiological methods were used to investigate the prevalence of dental fluorosis, and the levels of urine fluoride, serum fluoride, and drinking water fluoride in superficial well. The children were divided into six subgroups by the concentration of fluoride in drinking water: < 0.5 mg/L, 0.5-mg/L, 1.0-mg/L, 2.0-mg/L, 3.0-mg/L and > or = 4.0 mg/L.

RESULTSThere was significant dose-response relationship between the drinking water fluoride and the prevalence of dental fluorosis or the prevalence of defect dental fluorosis. The BMDLs (Benchmark Dose Lower Bound) were 1.01 and 1.30 mg/L, respectively. Urine fluoride and serum fluoride also had significant dose-response relationship to the prevalence of dental fluorosis or defect dental fluorosis. The correlation coefficient between drinking water fluoride and urine fluoride was 0.717, and it was 0.855 between drinking water fluoride and serum fluoride, and 0.617 between urine fluoride and serum fluoride.

CONCLUSIONSThe currently national standard of fluoride in drinking water in China is safe and reasonable. As a biological monitoring index, the levels of fluoride in serum may be more useful than that in urine in the control and prevention of endemic fluorosis.

Adolescent ; Child ; China ; epidemiology ; Environmental Monitoring ; Epidemiological Monitoring ; Female ; Fluorides ; analysis ; blood ; urine ; Fluorosis, Dental ; epidemiology ; Humans ; Male ; Prevalence ; Water Supply ; analysis ; standards

OBJECTIVETo summary the experience of the surgical comprehensive treatment of severe acute pancreatitis (SAP).

METHODSFrom July 1999 to December 2009, a total of 506 patients suffered SAP were admitted with a mean APACHE II score 12.8 ± 4.6. There were 270 male and 236 female, aged from 16 to 89 years, mean age 43 years. SAP patients were treated by the SAP treatment team which consisted of pancreatic specialized and multidisciplinary doctors. Two hundreds and thirty-four cases (46.2%) received non-operative treatment and 272 cases (53.8%) received surgical intervention.

RESULTSIn 506 cases, 445 patients were cured and 52 patients died (31 died in early stage, 21 died in later stage), 9 cases discharged automatically. The overall incidence of complication, overall mortality and overall curative rate were 29.4% (149/506), 10.3% (52/506) and 87.9% (445/506), respectively. The incidences of complication in non-operative group and in surgical intervention group were 27.8% (65/234) and 30.9% (84/272), respectively (P > 0.05). The mortality in non-operative group and in surgical intervention group were 9.4% (22/234) and 11.0% (30/272), respectively (P > 0.05). The curative rates in non-operative group and in surgical intervention group were 90.6% (212/234) and 85.7% (233/272), respectively (P > 0.05).

CONCLUSIONSPatients should be treated in ICU in the early phase of the disease when APACHE II score > 10. Pancreatic specialized and multidisciplinary team treatment, appropriate choice of timing, indication and procedure of surgical intervention and details of drainage are vital to the prognosis of SAP.

APACHE ; Acute Disease ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Pancreatitis ; mortality ; surgery ; Prognosis ; Retrospective Studies ; Survival Rate ; Young Adult