OBJECTIVETo study molecular epidemiology of norovirus (NV) infections, stool specimens collected from children with acute diarrhea were tested by TaqMan real-time reverse transcription polymerase chain reaction (RT-PCR) for the viral specific nucleic acid segments.

METHODSFecal samples from a total of 1260 children who had watery diarrhea seen from December 2006 to December 2007 in Guangzhou were analyzed by real-time RT-PCR. The primers and probes used for rapid detection and typing of NV strain target NV sequences were at the ORF1-ORF2 junction, a highly conserved region of the NoV genome. The positive specimens were determined by nested PCR and sequenced.

RESULTSTotally 257 specimens were positive for NV with a positive rate of 20.40%. Shedding of NV type GI was detected in 6.90%, type GII in 16.98% respectively, while the positive number of mixed infection with GI and GII was 44. Of the NV strains that were cloned and sequenced, GI was GI-3, GI-2 and GI-4 detected in positive specimens respectively; meanwhile, GII-4 was most commonly seen in genome II, followed by GII-3 and GII-7. In addition, the average age of children infected with NV was less than 2 years. An epidemic occurred during the winter and early spring (December through the next March).

CONCLUSIONNV was one of the important pathogens for acute diarrhea among children in Guangzhou, which suggested GII-4 was the prevalent strain.

Caliciviridae Infections ; epidemiology ; Child, Preschool ; China ; epidemiology ; Diarrhea ; epidemiology ; etiology ; virology ; Feces ; virology ; Humans ; Infant ; Molecular Epidemiology ; Norovirus ; classification ; genetics ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo study the effect of DPC4 gene expression on the growth of colon cancer cells and its mechanism.

METHODSExpression plasmid pcDNA3.1-DPC4 was constructed and transfected into the colon cancer cell line SW620 by use of lipofectamine gene transfer technique. DPC4 protein expression was detected by Western blot and immunocytochemistry. The effect of DPC4 gene on the growth of SW620 cells was monitored by population doubling time (PDT) and cloning efficiency. The influence of DPC4 expression on p21WAF1 transcription was investigated by RT-PCR to detect p21WAF1 mRNA.

RESULTSSuccessful expression of DPC4 protein was detected in the transfected SW620 cells. Compared with the control cells, PDT (74 h) of the DPC4 expressing cells was prolonged and the cloning efficiency (21%) decreased. In addition, the mRNA level of p21(WAF1) in DPC4 transfected cells was increased.

CONCLUSIONSOverexpression of DPC4 gene inhibits the growth of colon cancer in vitro, and induction of p21(WAF1) expression may be an important functional aspect of DPC4.

Carcinoma ; genetics ; metabolism ; pathology ; Cell Cycle Proteins ; biosynthesis ; genetics ; Cell Division ; Colorectal Neoplasms ; genetics ; metabolism ; pathology ; Cyclin-Dependent Kinase Inhibitor p21 ; DNA-Binding Proteins ; biosynthesis ; genetics ; Gene Transfer Techniques ; Genes, Tumor Suppressor ; Humans ; Smad4 Protein ; Trans-Activators ; biosynthesis ; genetics ; Transfection ; Tumor Cells, Cultured

OBJECTIVETo investigate the early prognostic values of arterial lactate and base excess (BE) in patients with paraquat poisoning.

METHODSSeventy-five patients with paraquat poisoning were divided into sudden death group (n = 10) who died within 24 h after admission, recent death group (n = 31) who died more than 24 h after admission, and survival group (n = 34). Arterial lactate and BE were measured on admission and at 24 h after admission. The prognostic values of arterial lactate and BE were analyzed.

RESULTSThe arterial lactate measured on admission was significantly higher in the sudden death group than in the recent death group and survival group (P < 0.01), but there was no significant difference in arterial lactate between the recent death group and survival group (P = 0.309). The BE measured on admission was significantly lower in the sudden death group than in the recent death group and survival group, and it was significantly lower in the recent death group than in the survival group (P < 0.01 or P < 0.05). At 24 h after admission, the recent death group had a significantly higher arterial lactate (P < 0.01) and a significantly lower BE (P < 0.01), as compared with the survival group. The logistic regression analysis showed that the two indices were significantly associated with prognosis (P < 0.01). On admission, the areas under the receiver operating characteristic (ROC) curve (AUCs) of arterial lactate and BE for predicting death were 0.692 and 0.787, respectively, and the cut-off values were 3.25 mmol/L and -1.75 mmol/L, respectively; the AUCs of arterial lactate and BE for predicting sudden death were 0.995 and 1, respectively, and the cut-off values were 7.1 mmol/L and -12.8 mmol/L, respectively. At 24 h after admission, the AUCs of arterial lactate and BE for predicting death were 0.743 and 0.822, respectively, and the cut-off values were 2.15 mmol/L and -5.55 mmol/L, respectively.

CONCLUSIONArterial lactate and BE have certain values in predicting the death, especially the sudden death, in patients with acute paraquat poisoning.

Adult ; Aged ; Arteries ; chemistry ; Female ; Humans ; Lactic Acid ; blood ; Male ; Middle Aged ; Paraquat ; poisoning ; Poisoning ; diagnosis ; Prognosis

OBJECTIVETo study the role of a pathologic niche inducing mouse embryonic stem cells (ESC) to express hepatic cell functions in vitro.

METHODSEmbryoid bodies were developed from 5 to 7 day hanging-drop culture of mouse ESC, and their dissociated cells were planted in three differential systems: nothing added; with 20 ng/ml hepatocyte growth factor (HGF); and 5% rat cholestatic serum plus 20 ng/ml HGF added. Their differentiation was observed with inverted microscopes daily, and their hepatic functions were analyzed against their synthesis of glycogen, triglycerides, albumin, and urea nitrogen, and by their staining of indocyanine green (ICG) and fluorescein diacetate (FDA).

RESULTSESC spontaneous differentiation was hardly being controlled to form three germ layers. HGF prompted the ESC to develop further into visceral endoderm and mesoderm (myocardium), but both of them only expressed a low level of hepatocyte-specific metabolic functions. With cholestatic serum added into the HGF-induced system, differentiated cells grew into similar angular cells, and had a higher level synthesis of glycogen, triglycerides, albumin and urea nitrogen with positive ICG and FDA staining.

CONCLUSIONSSpontaneous or HGF-induced ESC differentiation has only limited hepatic functions expressed. A pathologic niche in vitro induces ESC to develop into hepatic lineages, with a higher level of hepatic metabolic functions.

Animals ; Cell Differentiation ; physiology ; Cells, Cultured ; Cholestasis ; blood ; Culture Media ; pharmacology ; Embryo, Mammalian ; Hepatocytes ; cytology ; Mice ; Serum ; Stem Cells ; cytology

OBJECTIVETo investigate the correlation between major histocompatibility complex (MHC) class I chain-related gene A (MICA) gene alleles matching rates and graft rejection in small intestine, liver and kidney transplantation.

METHODSGenome DNA were extracted from blood samples or pathological sections collected from donors and recipients of living-related transplantation, included 4 cases of small bowel transplantation, 5 cases of liver transplantation and 6 cases of kidney transplantation. The correlation between MICA alleles matching rates and acute graft rejection was analyzed following 13 MICA alleles determination by polymerase chain reaction based on sequence-specific primers (PCR-SSP).

RESULTSHLA zygosity of all donors and recipients was confirmed to be half-matching. The recipients displaying higher matching rates of MICA alleles with donors showed lighter clinical and pathological rejection and longer survival time. On the contrary, recipients with lower matching rates of MICA alleles with donors showed severer clinical and pathological rejection and shorter survival time relatively.

CONCLUSIONMatching rates of MICA alleles has negative relevance to acute rejection, and positive relevance to survival time of recipients in small bowel, liver, and kidney transplantation.

Alleles ; Graft Rejection ; genetics ; immunology ; Histocompatibility Antigens Class I ; genetics ; Humans ; Intestine, Small ; transplantation ; Kidney Transplantation ; immunology ; Liver Transplantation ; immunology ; Living Donors ; Organ Transplantation

OBJECTIVETo evaluate the postoperative outcomes of preserving the left colonic artery during laparoscopic anterior resection for rectal cancer.

METHODSThe clinicopathologic data of 91 rectal cancer patients (pathologic Stage II) undergoing laparoscopic anterior resection was retrospectively analyzed. During the surgeries, the left colonic artery was preserved in 40 patients (preserved group) and ligated in 51 patients (unpreserved group). The operating time, intraoperative blood loss, time to first flatus and defecation, duration of postoperative abdominal distension and pain, number of retrieved lymph nodes, ileum fistulation and anatomical leakage rate were compared between the two groups.

RESULTSThe surgeries were completed in all the 91 patients laparoscopically without conversion. There was no intraoperative complications including rectal perforation, injury to vessel or ureter in either group. The operating time, blood loss and number of retrieved lymph nodes were similar between the groups (P>0.05). Three patients in preserved group and 5 in ligation group received preventive ileum fistulation due to low rectal cancer. Anatomical leakage occurred in three patients of unpreserved group. The average duration of postoperative abdominal distension and pain was 2.14∓0.35 days in preserved group and 3.15∓0.42 days in ligation group. The time to first flatus and defecation was 37.15∓12.62 h and 3.16∓0.52 days in preserved group and 62.25∓11.75 h and 4.25∓0.75 days in ligation group. Postoperative hospital stay was 4.54∓0.42 days in preserved group and 6.23∓0.51 days in ligation group. Total hospitalization cost in the two groups was 34 525.32∓1206.36 Yuan and 41 215∓1051.32 Yuan, respectively. Significant differences were found the in duration of postoperative abdominal distension and pain, postoperative hospital stay, and total cost between the two groups (P<0.05).

CONCLUSIONDuring laparoscopic anterior resection for rectal cancer, preserving the left colonic artery effectively ensures the blood supply to the anastomosis and the remaining descending colon to promote the recovery of the patients after surgery.

OBJECTIVETo investigate blood lead levels (BLLs) and influencing factors of BLLs among preschool children living in towns of Hunan Province.

METHODSA total of 2 044 preschool children (1,108 boys and 936 girls) from towns of 12 regions in Hunan Province were enrolled by a cluster sampling between September 2008 and June 2009. The average age of the children was 4.4 ± 1.1 years (range 2 to 6 years). BLLs were determined using the atomic absorption spectrographic method. The influencing factors of BLLs were investigated using a standard questionnaire and logistic regression analysis.

RESULTSThe mean BLLs of the children were 81.9 ± 34.5 μg/L. BLLs more than 100 μg/ L were noted in 482 children (23.58%). Of the 482 children, 472 (23.09%) showed BLLs of 100-199 μg/L and 10 (0.49%) showed BLLs ≥ 200 μg/L. There were significant differences in the prevalence of elevated BLLs (≥ 100 μg/L) among different age groups (P < 0.01). The prevalence of elevated BLLs in boys (28.99%) was significantly higher than that in girls (21.98%) (P < 0.01). There were significant differences in the prevalence of elevated BLLs in children from different regions (P < 0.01). The logistic regression analysis showed that the male (OR = 1.449, P < 0.01), father's occupational lead exposure (OR = 1.314, P < 0.01)and maternal frequent use of hair dyes (OR = 1.678, P < 0.05) were risk factor for elevated BLLs.

CONCLUSIONSThe prevalence of elevated BLLs is higher in preschool children living in towns of Hunan Province and is associated with a child's region and age. The male, father's occupational lead exposure and maternal frequent use of hair dyes are risk factor for elevated BLLs.

Child ; Child, Preschool ; China ; Female ; Humans ; Lead ; blood ; Logistic Models ; Male ; Sex Characteristics

In order to investigate whether Yinchenhao decoction (YCHD) attenuates hepatic fibrogenesis in the bile duct ligation (BDL) model via recovering and restoring the self-regulation and balance of the renin-angiotensin system (RAS), 33 specific-pathogen-free (SPF) male Sprague-Dawley rats with common BDL and scission were randomly divided into five groups as follows: G1, the sham group (n=4); G2, BDL 7-day group (n=5); G3, BDL+YCHD 430 mg/mL (n=8); G4, BDL+losartan 0.65 mg/mL (ARB group, n=8); G5, model group (BDL without any treatment, n=8). YCHD and losartan (10 mL·kg(-1)·day(-1)) were given by gastric gavage for 16 days following BDL in G3 and G4 groups, respectively. The effect of YCHD on liver fibrosis and the detailed molecular mechanisms were assessed by liver function including total bilirubin (TBIL), direct bilirubin (DBIL), indirect bilirubin (IDBIL), alanine aminotransferase (ALT), and aspartate aminotransferase (AST). Histological changes were observed by transmission electron microscopy (TEM) and Masson trichrome staining. Western blotting was used to detect the protein expression level of the renin-angiotensin system (RAS) components including angiotensin converting enzyme (ACE), angiotensin II type 1 receptor (AT1R), ACE2, angiotensin II (AngII) as well as transforming growth factor β1 (TGFβ1). The experimental data were analyzed by principle component analytical method of pattern recognition. The results showed that biochemically, serum TBIL, DBIL, IDBIL, ALT and AST levels were markedly increased following BDL as compared with the sham group (P<0.05). Serum TBIL, IDBIL and DBIL levels in G3 group were dramatically decreased as compared with G5 and G4 groups (P<0.05). Serum AST level in G3 was significantly lowered than in G5 group (P<0.05), but there was no significant difference in ALT among G3, G4 and G5 groups (P>0.05). Histologically, livers in G3 group showed less hepatocytes necrosis, less bile duct hyperplasia and less collagen formation than in G4 and G5 groups. The protein expression levels of ACE2, ACE, AngII, AT1R and TGFβ1 in G2, G3 and G4 groups were significantly higher than in sham group (P<0.05), and lower than in G5 group (P<0.05). However, the differences among G2, G3 and G4 groups were not significant (P>0.05). ACE2 protein expression in G3 group was significantly higher than in G2 group (P<0.05) and there was no significant difference in comparison with G4 group (P>0.05). Moreover, the protein expression of TGFβ1 in G3 group was significantly lower than in G5 and G4 groups (P<0.05). Our findings suggest that the antifibrotic effects of YCHD may be associated with the decreased classical RAS pathway components and TGFβ1 downexpression so as to recover and rebuild self-regulation of the RAS by elevating the protein expression of ACE2.
Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; pharmacology ; Gene Expression Regulation ; drug effects ; Liver ; drug effects ; pathology ; Liver Cirrhosis ; metabolism ; pathology ; prevention & control ; Liver Function Tests ; Losartan ; administration & dosage ; Male ; Rats ; Rats, Sprague-Dawley ; Renin-Angiotensin System ; drug effects

OBJECTIVETo compare the difference of effects on SiO(2)-induced alveolitis and early fibrosis between bone marrow-derived mesenchymal-like stem cells (BM-MSCs) and BM-MSCs transfected by pcDNA3.1-HGF and to explore the mechanism of this effects.

METHODSThe Primary BM-MSCs from Wistar male young rats were cultured and labeled by 4, 6-diamidino-2-phenylindole (DAPI). Fifty Wistar rats were randomly divided into 3 groups:model group (10 rats),which was administered with SiO(2) by the trache, the next day,injected PBS via the tail vein; BM-MSCs group (20 rats),which was administered with SiO(2) by the trache, the next day,injected with 1 ml suspension of BM-MSCs via the tail vein; pcDNA3.1-HGF plus BM-MSC group (20 rats),which was administered with SiO(2) by the trache, the next day,injected with 1 ml suspension of BM-MSCs transfected by pcDNA3.1-HGF via the tail vein. On the 14th and 28th days after treatment, half of the animals were sacrificed, respectively, and the lungs were harvested for frozen section to observe the cell marked by DAPI. HE staining under a fluorescent microscope, and to observe the pulmonary alveolitis and fibrosis by HE and Masson staining under a light microscope. Western blot assay was used to detect the expression of HGF in rat lungs. The expression levels of tumor necrosis factor-α (TNF-α) in pulmonary tissues were analyzed quantitatively by ELISA. The contents of HYP in pulmonary tissues were analyzed quantitatively by sample hydrolysis method.

RESULTSOn the 14th and 28th days after treatment, the scores of pulmonary alveolitis and early fibrosis in pcDNA3.1-HGF plus BM-MSCs group were 2.36 ± 0.17, 2.8 ± 0.14 and 0.1 ± 0.11, 1.16 ± 0.13, which were significantly lower than those (1.68 ± 0.17, 1.58 ± 0.31 and 0.54 ± 0.15, 1.36 ± 0.13) in BM-MSCs group, also which were significantly lower those (2.36 ± 0.17, 2.80 ± 0.14 and 0.64 ± 0.09, 1.84 ± 0.17) in model group (P < 0.05); On the 14th and 28th days after treatment, the TNF-α contents of pulmonary tissues in pcDNA3.1-HGF plus BM-MSCs group were 280.4 ± 23.11 and 249.78 ± 22.33 pg/mg, which were significantly lower than those (341.58 ± 35.34, 442.29 ± 36.76 pg/mg and 319.51 ± 17.84, 348.53 ± 33.95 pg/mg) in BM-MSCs and model groups (P < 0.05); On the 14th and 28th days after treatment, the HYP contents of pulmonary tissues in pcDNA3.1-HGF plus BM-MSCs group were 0.46 ± 0.04 and 0.65 ± 0.05 µg/mg, which were significantly lower than those (0.63 ± 0.04, 1.04 ± 0.07 µg/mg and 0.72 ± 0.60, 1.39 ± 0.60 µg/mg) in BM-MSCs and model groups (P < 0.05).

CONCLUSIONThe effects of BM-MSCs transfected by pcDNA3.1-HGF on suppressing pulmonary alveolitis and early fibrosis induced by SiO2 were better than those of BM-MSCs. The mechanism may be associated with the reduced pulmonary inflammation.

Animals ; Bone Marrow Cells ; cytology ; Hepatocyte Growth Factor ; genetics ; metabolism ; Male ; Mesenchymal Stromal Cells ; metabolism ; Pulmonary Fibrosis ; chemically induced ; prevention & control ; Rats ; Rats, Wistar ; Silicon Dioxide ; toxicity ; Silicosis ; prevention & control ; Transfection

BACKGROUND: Ankylosing spondylitis (AS) is a common chronic progressive rheumatic disease. The aim of this study was to explore factors influencing abnormal bone mineral density (BMD) in young and middle-aged patients with AS. METHODS: From July 2014 to August 2018, hospitalized patients with AS and health examinees in the health examination center of our clinics, ranging in age from 20 to 50 years, were monitored. The BMD of the lumbar spine and femoral neck of AS patients and those of a healthy control group were measured using dual-energy X-ray absorption. The BMDs of AS patients were compared with respect to age, course of disease, iritis, smoking habits, sex, height, weight, body mass index (BMI), medication use, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), platelet volume, platelet count, uric acid (UA), alkaline phosphatase (AKP), and calcium ion levels. Single-nucleotide polymorphisms (SNPs) related to BMD were screened using genome-wide association analysis. RESULTS: There was no statistical difference in the proportion of abnormal bone masses between the different body parts. The BMD of all bones in AS patients was lower than that in healthy controls (P < 0.05). Additionally, BMD was correlated with serum calcium and CRP in AS patients (P < 0.05), but not with age, platelet volume, platelet count, ESR, UA, AKP, height, weight, and BMI. The incidence of abnormal bone mass in AS patients was correlated with sex (P < 0.05), but not with medication use, iritis, or smoking. BMD of the lumbar spine in AS patients did not correlate linearly with the course of the disease, but BMD of the femoral neck correlated linearly with the course of the disease (P < 0.05). BMD was correlated with multiple SNPs in patients with AS. Lumbar BMD was correlated with rs7025373 and rs7848078. Femoral head BMD was correlated with 3:102157365, 3:102157417, rs1252202, rs1681355, rs3891857, rs7842614, and rs9870734, suggesting that genetic factors play a role in BMD in patients with AS. CONCLUSIONS The proportion of abnormal bone mass in AS patients was higher than that in healthy individuals of the same age. The factors related to BMD in patients with AS are gender, CRP, and blood calcium. The BMD of the femoral neck of AS patients decreases with the course of the disease, but BMD of the lumbar spine is not related to the course of the disease. BMD in AS patients is associated with multiple SNPs.
Absorptiometry, Photon ; Adult ; Bone Density ; Femur Neck ; Genome-Wide Association Study ; Humans ; Lumbar Vertebrae ; Middle Aged ; Spondylitis, Ankylosing/genetics* ; Young Adult