Objective To explore the association of Crohn's disease (CD) with T cell immunoglobulin and mucin domain 3 (Tim-3) gene polymorphisms in patients of Zhejiang Han population in China.Methods A total of 308 CD patients and 573 age-and sex-matched healthy controls were enrolled in our study.Two single nucleotide polymorphisms (SNPs) of Tim-3 (rs1036199 and rs10515746) were examined by the improved multiple ligase detection reaction technique (iMLDR).Analyses of linkage disequilibrium and haplotype were also performed by Haploview 4.2 software in all study subjects.Results In general,the allele and genotype frequencies of Tim-3 (rs1036199 and rs10515746) were not statistically different between CD patients and the controls (all P ＞0.05).According to the Montreal Classification,CD patients were divided into different subgroups.The variant allele (C) and genotype (AC + CC) of rs1036199 were more frequent in CD patients with penetrating diseases than in the controls (10.4％ vs 1.7％,P =0.002;20.8％ vs 3.5％,P =0.023).Similar conclusions were also drawn for the variant allele (A) and genotype (CA + AA) of rs10515746 in patients with penetrating diseases when compared with the controls (10.4％ vs 2.2％,P =0.000;20.8％ vs 4.2％,P =0.033,respectively).The two SNPs of Tim-3 were in strong linkage disequilibrium (D'=1.0,r2 =0.928).The haplotype (AC) formed by their wild-type alleles (A) and (C) was decreased in patients with penetrating CD compared with the controls (89.6％ vs 98.3％,P =0.000).However,the haplotype (CA) formed by their variant alleles was more frequent in patients with penetrating CD than in the controls (10.4％ vs 1.6％,P =0.000).Conclusions Tim-3 (rs1036199 and rs10515746) variations might be correlated with the enhanced risk of penetrating diseases in CD patients.Furthermore,the haplotype (AC) and (CA) formed by the two SNPs might be a protective and a risky factor for penetrating CD respectively.

Objective:To systematically evaluate the efficacy and safety of neoadjuvant chemoradiotherapy (NCRT) plus surgery versus neoadjuvant chemotherapy (NCT) plus surgery in the treatment of advanced esophageal squamous cell carcinoma.Methods:Clinical controlled trials of comparing the treatment of NCRT plus surgery with NCT plus surgery for esophageal squamous cell carcinoma were electronically searched from the databases including PubMed, The Cochrane Library, EMbase, CBM, CNKI, WanFang and VIP from the inception of databases to January, 2019. Two reviewers independently screened the literatures, extracted data and assessed the risk of bias of the included studies. And then, a meta-analysis was performed by using RevMan 5.3 software.Results:A total of 8 clinical control studies were included, including 995 patients with esophageal squamous cell carcinoma. Meta-analysis results showed that compared with the NCT group, the R 0 resection rate was significantly higher ( OR=2.14, 95% CI: 1.03-4.45, P=0.040) and the pathological complete response (pCR) rate was significantly higher ( OR=4.19, 95% CI: 1.71-10.28, P=0.002) in the NCRT group. The incidence of postoperative complications ( OR=1.37, 95% CI: 0.76-2.48, P=0.300) and the risk of perioperative death ( OR=1.28, 95% CI: 0.58-2.83, P=0.54) were not significantly different between two groups. The long-term survival of patients with esophageal squamous cell carcinoma in the NCRT group was significantly better compared with that in the NCT group ( HR=0.77, 95% CI: 0.64-0.92, P=0.005). Conclusions:Compared with NCT plus surgery for advanced esophageal squamous cell carcinoma, NCRT plus surgery has higher R 0 resection rate and pCR rate, does not significantly increase the risk of perioperative complications or perioperative death, and significantly improves the long-term survival of esophageal squamous cell carcinoma patients.

Objective To investigate the daily total fluoride intake in relating to the prevalence of skeletal fluorosis in two villages in Jiangsu Province,in order to provide the scientific evidences for the control and prevention of endemic fluorosis.Methods Adults sampled from a high-fluoride Village,Wamiao,and a low-fluoride Village,Xinhuai,were surveyed in this study according to the fluoride concentration in their household shallow well.The average daily total fluoride intake from difierent sources and the skeletal fluorosis were investigated in each subject.Then the subjects from two villages were allocated into five subgroups(＜2.00,2.00～,3.00～,4.00～,≥5.00 mg/d),the relation fluoride intake and prevalence of osteofluorosis was analyzed.Results The prevalence of skeletal fluorosis in Wamiao Village was 31.06%(41/132),but no skeletal fluorosis case(0/35)was found in Xinhuai Village.According to the daily total fluoride intake,subjects with higher daily total fluoride intake tended to associated with a higher prevalence of skeletal fluorosis in a significant dose-response relationship(regression equation:y=2.624-6.855x+3.424x2:r=0.997).The benchmark dose lower limitation of daily total fluoride intake with 95% confidencewas 2.50 mg/d calculated according to this dose-response relationship,the reference dose(RfD)was 2.50 mg/d.In Wamiao Village a significant difference was also found between daily total fluoride intake in 41 subjects[(5.09±1.20)mg/d]with X-ray detectable skeletal fluorosis and in 91 subjects[(3.08±1.12)mg/d]without X-ray detectable skeletal fluorosis(t=-9.32,P＜0.01).Conclusions These findings indicate that the daily total fluoride intake has a significant dose-response relationship with the prevalence of skeletal fluorosis in an endemic fluorosis area associated with high-fluoride drinking water;and the RfD in this study was lower than that in the national standard of"Chinese hygienic standard for daily total fluoride intake(WS/T 87-1996)"(3.50 mg/d).

OBJECTIVETo observe therapeutic effects of bird-pecking moxibustion in children of hand, foot and mouth disease, and to analyze the mechanism.

METHODSSeventy-five children of hand, foot and mouth disease were randomly divided into 3 groups, a combined moxibustion and medicine group (n = 22), a Chinese medicine group (n = 29), and a western medicine group (n = 24). The combined moxibustion and medicine group was treated with bird-pecking moxibustion combined with routine western medicine, the Chinese medicine group with oral administration of Chinese medicine and routine western medical therapy, and the western medicine group with routine western medicine. After treatment of 7 days, the therapeutic effects on skin rash, oral herpes, constipation or loose stool, dyspepsia and anorexia, etc. were comprehensively assessed, and their therapeutic effects were compared.

RESULTSThe total effective rate was 95.5% in the combined moxibustion and medicine group, 86.2% in the Chinese medicine group, and 83.3% in the western medicine group, the former being significantly better than those of the other two groups (both P < 0.05). The combined moxibustion and medicine group was significantly better than the other two groups in the relieving time of skin rash, oral herpes, constipation or loose stool, dyspepsia and anorexia, etc. (all P < 0.05)

CONCLUSIONThe combined moxibustion and medication can effectively improve symptoms of the digestive tract, shorten duration of disease, reduce pain in the patient with hand, foot and mouth disease.

Child ; Child, Preschool ; Combined Modality Therapy ; Drug Therapy ; methods ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Female ; Hand, Foot and Mouth Disease ; pathology ; therapy ; Humans ; Infant ; Male ; Moxibustion ; methods ; Treatment Outcome

OBJECTIVETo investigate the effects of carnosic acid (CA) on reversal of the multidrug resistance (MDR) of human leukemia cell line K562/A02 and its mechanism.

METHODSMTT assay was used to determine the sensitivity of K562/A02 cells to adriamycin (ADM) pre-and post-treated with CA. Flow cytometry (FCM) and laser scanning confocal microscopy (LSCM) were used to measure intracellular fluorescence intensity and concentration of ADM respectively. The expression level of mdr1 was detected by semi-quantitative RT-PCR. P-glycoprotein (P-gp) expression was detected by FCM and Western blot.

RESULTSCA decreased IC(50) of ADM in K562/A02 cells from 16.31 µg/mL to 1.35 µg/mL, being a 12.08-fold decrease. The intracellular ADM fluorescence intensity of K562/A02 was increased from 17.05 to 60.53 after treated with CA (P < 0.01). In living K562/A02 cells, after treated with CA, the diffuse distribution of intracellular ADM was recovered in both nuclear and cytoplasm, and the concentration of intracellular ADM increased from 4.93µg/mL to 15.43µg/mL. RT-PCR assay showed that CA inhibited the expressions of mdr1 mRNA in K562/A02 cells (P < 0.01). Mean fluorescence intensity of P-gp detected by FCM in CA-treated K562/A02 was decreased to 22.80 as compared with that in untreated K562/A02 cells (44.40, P < 0.05).

CONCLUSIONCA can reverse the MDR of K562/A02 cells in vitro. The mechanism may be associated with down-regulation of mdr1 and inhibition of P-gp function.

ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Humans ; K562 Cells

OBJECTIVETo compare the sensitivity of Brown Norway rats (BN) with Guinea pigs (GP) as allergen assessment animal models.

METHODBN rats and GP were randomly assigned to 1 control group, 2 Bovine serum albumin group (BSA), respectively. Animals in BSA groups of BN rats and GPs were sensitized by intraperitoneal injection of 0.6% BSA 1 ml on day 1, 3, 5, respectively, and irritated by intravenous injection of 2.4% BSA 1 ml on day 7 and day 14 after the last sensitization, while the same volume of normal saline was given to control group on each time point mentioned above. The allergic reactions were scored within 1 h after each irritation treatment, and the sera of both BN rats and GPs were collected to detect IgE concentration by using ELISA. The sera were also applied for passive cutaneous anaphylaxis test (PCA test) in SD rats.

RESULTNo obvious allergic reactions were observed in BSA group of GPs after each irritation treat, however, the score of allergic response in BSA group of BN rats was evidently higher than that in control group after first irritation. PCA test by using sera from BSA group of BN rats after both irritations showed the strong positive result characterized as large amount of subcutaneous effusions of Evans blue in SD rats, however, the sera from BSA group of GP were negative in PCA test. Serum IgE concentration did not increase after each irritation in BSA group of both BN rats and GP.

CONCLUSIONBN rats were more sensitive than GPs on initiative systemic anaphylaxis test and passive cutaneous anaphylaxis test. Meanwhile, BN rats has an advantage in experimental treatment compared with Guinea pigs.

Allergens ; administration & dosage ; toxicity ; Anaphylaxis ; chemically induced ; Animals ; Guinea Pigs ; Hypersensitivity ; etiology ; Male ; Models, Animal ; Rats ; Rats, Inbred BN ; Rats, Sprague-Dawley

OBJECTIVETo investigate the synergistic effects of carnosic acid (CA) with arsenic trioxide (As₂O₃) on proliferation and apoptosis in HL-60 human myeloid leukemia cells, and the major cellular signaling pathway involved in these effects.

METHODSHL-60 cellular proliferation was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) analysis. Cell cycle distribution and apoptosis were monitored by flow cytometry. The activation of casepase-9, Bcl-2-associated agonist of cell death (BAD), p-BAD, p27, phosphatase and tensin homolog deleted on chromosome ten (PTEN), Akt, p-Akt was assessed by Western blot analysis. The expression of PTEN mRNA was tested by reverse transcription polymerase chain reaction (RT-PCR) analysis.

RESULTSCA reduced HL-60 cell viability in a dose- and time-dependent manner, and induced G1 arrest and apoptosis. Moreover, CA upregulated PTEN expression, blocked the Akt signaling pathway, subsequently inhibited phosphorylation of BAD, reactivated caspase-9, and elevated levels of p27. CA also augmented these effects of As₂O₃.

CONCLUSIONCA might be a novel candidate of the combination therapy for leukemia treatment; these effects were apparently associated with the modulation of PTEN/Akt signaling pathway.

Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Base Sequence ; Blotting, Western ; Cell Cycle ; drug effects ; DNA Primers ; Diterpenes, Abietane ; pharmacology ; Drug Synergism ; HL-60 Cells ; Humans ; Leukemia, Myeloid, Acute ; metabolism ; pathology ; Oxides ; pharmacology ; PTEN Phosphohydrolase ; metabolism ; Plant Extracts ; pharmacology ; Proto-Oncogene Proteins c-akt ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; drug effects

Objective To evaluate the efficacy and safety of intravesical instillation of sodium hya- luronate in relieving the symptoms of refractory non-bacterial cystitis.Methods Totally,20 patients(1 man and 19 women;mean age,47 years;age range,22-68 years)with refractory non-bacterial cystitis were included.Their disease course ranged from 1 month to 30 years.Of them,16 patients had the disease for more than 1 year.Instillation of sodium hyaluronate(50 ml solution of 40 mg HA)was performed intravesically in them for a total of 12 weeks(once a week for the first 4 weeks and once every 4 weeks for the succeeding 8 weeks).Follow-up visit ended at 16 weeks,when the patients filled in the symptom assessment scores(VAS on pain,frequency and urgency)according to the subjective evaluation.Therapeutic results were classified as(1)total response:symptom disappearance or relief by at least 90% of the symptom scores;(2)partial response:symptom relief by 50%-89%;(3)mild response:symptom relief by 49% or less;(4)no re- sponse:no improvement in symptom scores.Results Nineteen patients completed the 16-week follow-up with only 1 withdrawing.The average general symptom score decreased from 22.32?5.53 to 9.47?5.88 (P＜0.001);frequency score from 8.21?1.75 to 3.89v2.31(P＜0.001);urgency score from 7.47?2.14 to 3.37?2.14(P＜0.001);pain score from 6.63?3.47 to 2.21?2.74(P＜0.001).After 16 weeks,overall positive response rate(total+partial response)was 53%(10/19).No serious adverse effect occurred except for urethral irritation(25%,5/20),worsening of distending pain of the bladder(5%,1/20) and vertigo(15%,3/20)caused by the procedure itself.Conclusions For patients suffering from refrac- tory non-bacterial cystitis,intravesical instillations of sodium hyaluronate can significantly relieve the symp- toms and is safe and of good patient compliance.

The industry of germ-free animals has been a hot spot in research along with the rapid development of studies on the relationship between microbiota and host diseases. Because it is pathogen?free, and the high degree of simi?larity in anatomy, physiology, pathogenesis to humans, germ?free pig is considered a clinical relevant model to be widely used in life science research. Based on the current state of research of germ?free pig cultivation at home and abroad and the experimental studies carried out in our laboratory as well, this article gives a simple discussion on germ?free technique of domestic pigs.

OBJECTIVETo investigate the epidemiology of late-onset hypogonadism (LOH) in old and middle-aged males in the rural area of Southern China.

METHODSUsing the age-stratified sampling method, we conducted a questionnaire investigation on androgen deficiency in aging males (ADAM), aging male symptoms (AMS) and IIEF-5 among 996 males aged 40 -80 years in a rural community of Jiashan county, Zhejiang Province from April to October 2012. We also determined the concentrations of serum total testosterone (TT), serum sex hormone binding globulin (SHBG) and serum albumin (ALB), detected the levels of free testosterone (cFT) and bio-available testosterone (Bio-T) by Vermeulen formula, and measured the volumes of the prostate and testis by ultrasonography.

RESULTSThe mean age of the males was 56.22 +/- 8.82 years. The positive rates of LOH were 62.86% and 23.05% based on ADAM and AMS, respectively, and the incidence of erectile dysfunction (ED) was 68.83%. There were significant differences among different age groups in the levels of luteinizing hormone (LH), SHBG, cFT and Bio-T, but not in TT concentration.

CONCLUSIONOurs was the first survey on the epidemiology of LOH among old and middle-aged males in the rural area of China. The incidence of LOH in the rural community of Zhejiang Province was lower than that in the urban areas reported in other studies, but the positive rate of ED showed no significant difference.

Adult ; Age of Onset ; Aged ; Aged, 80 and over ; China ; epidemiology ; Erectile Dysfunction ; epidemiology ; Humans ; Hypogonadism ; epidemiology ; Incidence ; Luteinizing Hormone ; blood ; Male ; Middle Aged ; Rural Population ; Surveys and Questionnaires ; Testosterone ; blood