Viable but non-culturable state in bacterial cells as received uncultured microorganism has be- come the fundamental research issues in medical microbiology, epidemiology, general microbial ecology and sanitation quarantine since it was put forward in 1982. When in this state, bacterium are no longer able to grow and form colonies on conventional culture media, but maintain their patheogenicity, may become latent infection escaping detection to threaten enviroment and human security. With the development of modern molecular biology technique and metagenomics, it provide new research method and opportunity for uncul- tured microorganism in the environment. Recently, with the application of the metagenome technique, ge- netic fingerprinting technical etc, it becomes more and more popular. Simultaneously, along with this state become maturity in the laboratory. It provides a new research route for development and exploitation the uncultured microorganism.

Objective:To investigate the value of magnetic resonance cholangiopancreatography(MRCP)in the diagnosis of pancreas divisum by comparing with endoscopic retrograde cholangiopancreatography(ERCP).Methods:The MRCP and ERCP images of 8 patients with pancreas divisum were retrospectively analyzed.The diagnostic accuracy and findings by MRCP were compared with those by ERCP.Results:MRCP had a diagnostic accuracy of 87.5%(7/8)based on the result of ERCP.ERCP displayed the dominant dorsal pancreatic ducts in all 8 cases and ventral pancreatic ducts in 6 cases;MRCP also displayed the dominant dorsal pancreatic ducts in all 8 cases,but the ventral pancreatic ducts only in 3 cases.Conclusion:As a non-invasive technique,MRCP has important clinical value in the diagnosis of pancreas divisum.

Objective To determine the effects of 1320 nm non-ablative laser on the proliferation of human dermal fibroblasts,and the secretion of basic fibroblast growth factor(bFGF)and transforming growth factor-?1(TGF-?1)in vitro.Methods Human dermal fibroblasts were cultured,and irradiated three times by 1320 nm laser at a dose of 15,20 and 24 J/cm~2,respectively.The levels of bFGF and TGF-?1 were examined by ELISA at 0,24,48 and 72h after the irradiation.The number of fibroblasts before and after irradiation were determined.Results The number of fibroblasts and the secretion of bFGF both in- creased after the irradiation at the doses of 20 J/cm~2 and 24 J/cm~2(P

OBJECTIVETo explore the correlation of the gene polymorphisms of Toll-like receptor 2 ( TLR2) and TLR4 with the susceptibility and recurrence of condyloma acuminatum (CA).

METHODSUsing Snapshot, we detected the gene polymorphisms of TLR2 597(T/C), 1350(T/C), 15607(A/G), and 2258(G/A) and TLR4 896(A/G) and 1196(C/T) in the peripheral blood of 140 CA patients and 105 HPV-negative controls. We made comparisons between the CA patients and controls as well as between the cases of recurrent CA and those of non-recurrence at 6 months after treatment.

RESULTSThere were 72, 48, and 20 cases of genotype TT, TC, and CC of TLR2 597 (T/C), respectively, in the CA patients, as compared with 71, 31, and 3 cases in the controls. The gene frequency of mutant C was 31. 43% in the patients, significantly higher than 17.62% in the controls (χ2 = 12.04, P < 0.01), and it was 38.68% in the recurrent cases, remarkably higher than 27.01% in the non-recurrent cases (χ2 = 4.16, P < 0.05). There were 74, 49, and 17 cases of genotype TT, TC, and CC of TLR2 1350( T/C), respectively, in the CA patients, as compared with 73, 29, and 3 cases in the controls. The gene frequency of mutant C was 29. 64% in the patients, significantly higher than 16. 67% in the controls (χ2 =11.05, P < 0.01), and it was 36.79% in the recurrent cases, markedly higher than 25. 29% in the non-recurrent cases (χ2 = 4.18, P < 0.05). There were 44, 66, and 30 cases of genotype AA, AG, and GG of TLR2 15607(A/G), respectively, in the CA patients, as compared with 26, 58, and 21 cases in the controls. There was no significant difference in the gene frequencies of mutant G between the two groups (χ2 = 0.33, P > 0.05). No mutant genes of TLR2 2508 (G/A) or TLR4 896(A/G) and 1196(C/ T) were detected in either the CA patients or the controls. Linkage disequilibrium analysis showed a tight linkage between TLR2 597 (T/C) and 1350(T/C) (D' = 1, r2 = 0.93).

CONCLUSIONTLR2 597(T/C) is tightly linked to 1350(T/C), which is correlated with both the susceptibility and the recurrence of condyloma acuminatum.

Aged ; Case-Control Studies ; Condylomata Acuminata ; genetics ; Gene Frequency ; Genetic Linkage ; Genetic Predisposition to Disease ; Genotype ; Humans ; Polymorphism, Genetic ; Recurrence ; Toll-Like Receptor 2 ; genetics ; Toll-Like Receptor 4 ; genetics

BACKGROUNDThe purpose of this study was to evaluate the efficacy of topical melagenine for repigmentation in child patients with vitiligo on the scalp.

METHODSTwenty-two child patients with vitiligo on the scalp were treated with 1.2 mg/ml aqueous melagenine in combination with 20 minutes of infrared exposure twice daily.

RESULTSIn 4 patients (18.2%), melagenine treatment in combination with infrared exposure led to complete recovery; in 6 patients (27.3%), treatment was shown to be effective; in 8 patients (36.3%), treatment led to improvements in patient condition; and only 4 patients (18.2%) showed no response after 1 - 2 treatment sessions. The general effective rate of melagenine-infrared combination treatment was 45.5% for the children with vitiligo on the scalp, and treatment was accompanied by minimal side effects.

CONCLUSIONMelagenine may be efficacious and a safe treatment option for childhood vitiligo affecting the scalp.

Administration, Topical ; Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infrared Rays ; therapeutic use ; Lipoproteins ; administration & dosage ; Male ; Scalp Dermatoses ; drug therapy ; Vitiligo ; drug therapy

Objective To investigate DNA methylation in the promoter region,mRNA transcription and protein expression of glutathione-S-transferases-P1 (GSTP1) gene and their relation with arsenism.Methods In endemic coal-pollution-borne arsenism area,Jiaole village of Xinren county,Guizhou province,according to the diagnostic criteria of endemic arsenism(WS/T 211-2001),123 cases with endemic arsenism were selected and divided into three groups (mild arsenism group:42 cases,moderate arsenism group:41 cases and severe arsenism group:40 cases).Forty seven residents were selected as controls in a village about 12 km away from the endemic arsenism area.With the informed consent principle,peripheral blood of all respondents was collected in order to analyze DNA methylation and check mRNA.DNA methylation of GSTP1 gene promoter region in peripheral blood was assayed by PCR,and GSTP1 mRNA expression was assayed using real-time quantitative PCR.In addition,other cutaneous specimens originated from 53 cases with arsenism that accepted surgical treatment voluntarily were taken.Of these specimens,general pathological changes were 28 cases,precancerous 20 cases and cancerous 5 cases.Skin tissues of 15 cases of non-tumor surgery patients without abnormal pathological changes were as control group.GSTP1 protein expression in the skin tissue was detected using immunohistochemistry (IHC).Results Among different groups of arsenic poisoning,the positive rate of DNA methylation of GSTP1 gene was 28.57％(12/42) in the mild group,57.10％ (23/41) in the moderate group and 65.00％ (26/40) in the severe group.Compared with the control group (6.38％,3/47),the difference was statistically significant (x2 =7.792,26.000,33.412,all P ＜ 0.01).Among different groups of arsenic poisoning diagnosed by dermapathology,the positive rate of DNA methylation of GSTP1 gene was 21.43％(6/28) in the general pathological change group,50.00％(10/20) in the precancerous group and 80.00％(4/5) in the cancerous group.Compared with the control group(6.67％,1/15),the difference was statistically significant (x2 =3.562,7.468,10.756,all P ＜ 0.05).It showed that the positive rate of DNA methylation of GSTP1 gene increased with aggravation of the disease and dermatic lesion of arsenism (tendency x2 =38.239,x2 =13.659,all P ＜ 0.01).Compared with the control group(0.184 26),the expressions of GSTP1 mRNA in peripheral blood in moderate (0.087 77) and severe arsenic poisoning groups (0.056 93) were significantly reduced(all P ＜0.01),and that of severe group was significantly lower than that of the moderate group (P ＜ 0.01) ; compared with the control group(0.338 45) and the general lesion group(0.276 74),GSTP1 mRNA expression was significantly reduced in precancerous lesion group(0.104 81) and cancerous group(0.043 70),in which the cancerous group was significantly lower than that of the precancerous lesions.The difference of skin tissue GSTP1 protein expression rate between groups was statistically significant (x2 =20.948,P ＜ 0.05),in which the difference between the precancerous lesion group(65.00％,13/20),the cancer group (40.00％,2/5) and the control group(100.00％,15/15)was statistically significant (x2 =12.183,11.778,P ＜ 0.01).Spearman correlation analysis showed that the degree of skin lesion and the level of GSTP1 protein expression was negatively correlated (r =-0.520,P ＜ 0.05).Groups were divided according to DNA methylation of GSTP1 gene,and the mRNA and protein expression of GSTP1 in methylation group(0.038 40,57.14％) was significantly lower compared with that of unmethylated group(0.187 07,95.74％; Z =9.032,x2 =23.134,all P ＜ 0.01).Conclusions Arsenism may lead to DNA methylation of human GSTP1 gene promoter region,thereby inhibiting expression of mRNA and protein.GSTP1 gene plays an important role in arsenism or carcinogenic process.

Objective To explore the effect of adenovirus carrying pigment epithelium-derived factor(ADV-PEDF)gene treatment on retinal neovascularization of mice with retinopathy of prematurity (ROP).Methods Sixty 7 d C57BL/6J mice were put into the environment with (750?5)mL/L oxygen for 5 days and returned to normal environment to establish animal models of ROP.The eyes in experimental group received an intravitreal injection of 1 ?L of ADV-PEDF,and the same volume of ADV-LacZ was injected into the eyes of mice in control group.The ADPase histochemical staining was used for retinal flatmount to observe changes of retinal vessels.The inhibitory effects of PEDF on retinal neovascularization were evaluated by counting the endotheliocyte nuclei of new vessels extending from retina to vitreous in the tissue-slice.The expression of PEDF in retina were detected by Western blotting.Results The vessels from optic disc were very thin and distorted in eyes of control group in retinal flatmount.There were avascular area around optic disc and neovascular trufts beside avascular area.Compared with control group,regular distributions and no conspicuous avascular area were found in eyes of experiment group in retinal flatmount.The number of the endotheliocyte nuclei of new vessels extending from retina to vitreous was less in the eyes of the experiment group than that in control group (P

Acupuncturist makes herbal cakes with traditional manual way, with such disadvantages as slow in making, varying in thickness and size of the cake. When the patients are treated with medical cake-separated moxibustion, they will be unevenly affected by the heat and the patient easily suffers from burning. These hinder clinically wide application of cake-separated moxibustion. With practice of many years, the authors design and make a kind of manual cake-shaping apparatus which can rapidly and conveniently make uniform medical cake, with simple technique, ingenious structure and normal material.
Equipment Design ; Humans ; Moxibustion ; instrumentation ; methods

There have been very few studies on the effect of Gualou Xiebai Banxia decoction combined with Xuefu Zhuyu decoction in inhibiting apoptosis in myocardial ischemial injury caused by coronary heart disease. In this experiment, Gualou Xiebai Banxia decoction combined with-Xuefu Zhuyu decoction were used to intervene the miniature swine phlegm and blood stasis type coronary heart disease model, in order to observe the effect of the combined prescription on the myocardial apoptosis and the expressions of Bcl-2, Bax, Caspase-3, Caspase-9 in the model. Totally 15 Chinese experimental miniature swine were adopted and randomly divided into the control group, the model group and the phlegm and stasis-treating group. The model group and the stasis-treating group were fed with high fat diets for two weeks, intervened with the coronary artery injury and then given drugs and high fat diets for eight weeks. The control group was fed with ordinary diets for 10 weeks, without the coronary artery injury. After the experiment, myocardia at the juncture of infracted areas were collected and made into formalin-fixed paraffin sections. The TDT-mediate dUTP nick end labeling (TUNEL) assay was used to detect the myocardial apoptosis. The immunohistochemistry (IHC) technique was applied to detect Bcl-2, Bax, Caspase-3, Caspase-9 levels in myocardial tissues. According to the findings, the apoptosis indexes (AI) for the control group, the model group and the phlegm and stasis-treating group were 0.92%, 27.68%, 17.28%, respectively. The AI of the phlegm and stasis-treating group was significantly lower than that of the model group (P < 0.01). Compared with the model group, the phlegm and stasis-treating group showed significantly higher Bcl-2 protein expression (P < 0.01) and lower Bax, Caspase-3 and Caspase-9 protein expressions (P < 0.01). In conclusion, Gualou Xiebai Banxia decoction combined with Xuefu Zhuyu decoction have a significant protective effect against the myocardial apoptosis in miniature swine phlegm and blood stasis type coronary heart disease model.
Animals ; Apoptosis ; drug effects ; Caspases ; metabolism ; Coronary Disease ; drug therapy ; Disease Models, Animal ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Male ; Medicine, Chinese Traditional ; Myocardium ; pathology ; Phytotherapy ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Swine ; Swine, Miniature ; bcl-2-Associated X Protein ; analysis

Objective To investigate the expression of VIT-1 gene in melanocytes of patients with vitiligo, and to analyze the difference of its sequence. Methods The skin from the foreskins of healthy men by circumcision and from the non-lesional area on the buttocks of 5 patients were digested by dispase, then the epidermis and dermis were separated, and the melanocytes were isolated. Then we cultured the melanocytes from the controls in TICVA medium and those from the patients in TICVA medium supplemented with basic fibroblast growth factor (bFGF) and endothelin-1 ( ET-1). The expression of VIT-1 gene was measured by RT-PCR, the full-length cDNA of VIT-1 ORF was cloned and sequenced, and sequence difference was analyzed by CLUSTAL W ( 1.83 ) software. Results The expression levels of VIT-1 gene were significantly lower in melanocytes from the patients than in those from the controls. An 81 bp-intron was found in the VIT-1 ORF. VIT-1 was a fragment of FBXO11, located at its 3' end. Conclusion VIT-1 gene is not a new gene, but a fragment of FBXO11, and a member of F-box protein family.