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Objective:To explore the effect of probiotics Lactiplantibacillus plantarum(LP) WCFS1 by gavage on acute necrotizing pancreatitis (ANP) and associated ileum injury in mice. Methods:Twenty-four healthy male mice were gavaged with broad-spectrum antibiotics for 3 weeks to establish microbiota-depleted mice, and then randomly divided into control group (CON), ANP model group (ANP), LP gavage group (LP) and LP gavage and ANP induced group (LP+ ANP) , with 6 mice in each group. Mice in LP and LP+ ANP group were treated by gavage of LP (1×10 9 CFU/ml, 0.2 ml/day per mouse) for 1 week, while CON and ANP were gavaged with sterile phosphate buffered saline for 1 week instead. The ANP model was induced by intraperitoneal injection with caerulein (100 μg/kg) for 10 times with 1-hour interval between two injections and the 10th injection with lipopolysaccharide(LPS) 5 mg/kg intraperitoneally, and the mice were sacrificed 2 h later. Levels of LP in stool and ileal mucosa were detected by real-time PCR; the pancreas and ileum were collected for pathological examination to observe the extent of tissue inflammation and to score the pathology. Serum amylase activities were determined by enzymatic kinetic chemistry; serum inflammators levels and intestinal permeability were detected by ELISA; levels of inflammators in pancreatic and ileal tissues were detected by real-time PCR; ileal tight-junction proteins (occludin, claudin-1 and ZO-1) were measured by immunofluorescence staining. Results:LP levels in the stool and ileal mucosa of mice were significantly increased after LP gavage, and the differences were statistically significant (913.30±39.12 vs 2.39±1.39, 23.11±0.50 vs 1.38±0.28, all P value <0.05). The pathological scores of pancreatic tissue of CON, LP, ANP and LP+ ANP group were (0.26±0.41), (0.17±0.26), (8.55±0.46) and (6.30±0.45); the serum amylase activities were (219.70±19.73), (217.60±11.30), (2896.24±98.32) and (1837.13±131.60)U/L, IL-1β were (0.87±0.28), (1.4±0.85), (67.41±6.45) and (36.33±5.65)pg/ml, IL-6 were (0.74±0.27), (0.16±0.16), (280586.12±39163.92) and (107912.75±31283.47)pg/ml, IL-10 were (35.52±5.27), (50.99±15.34), (2008.45±184.83) and (3070.35±403.71)pg/ml; the expression level of pancreatic IL-1β mRNA was 1.42±0.39, 0.95±25, 20.53±0.50 and 10.69±1.01, IL-6 mRNA was 1.31±0.44, 0.93±0.023, 21.97±1.71 and 11.54±1.75, IL-10 mRNA was 0.93±0.14, 0.75±0.15, 0.99±0.21 and 1.76±0.19; there was no significant difference between LP and CON group, and pancreatic pathological scores, serum amylase、IL-1β and IL-6 levels, and the expression level of pancreatic IL-1β and IL-6 mRNA were significantly decreased in LP+ ANP group compared with those in ANP group, while serum IL-10 levels and the expression level of pancreatic IL-10 mRNA were significantly increased compared with ANP group, and all the differences were statistically significant (all P values <0.05). The pathological scores of ileal tissue of CON, LP, ANP and LP+ ANP group were 0, 0, (3.17±0.41) and (1.67±0.52); the levels of serum DAO of CON, LP, ANP and LP+ ANP group were (0.03±0.03), (0.02±0.02), (0.50±0.05) and (0.49±0.06)ng/ml; LPS levels were (2.75±0.35), (3.74±0.28), (7.19±0.92) and (5.88±0.38)ng/ml; the expression level of ileal IL-1β mRNA was 1.21±0.20, 1.17±0.09, 1.81±0.25 and 1.63±0.21; IL-6 mRNA was 1.01±0.29, 2.83±0.42, 54.45±8.50 and 16.87±4.42; IL-10 mRNA was 1.12±0.41, 6.09±2.51, 11.65±1.47 and 29.86±2.93. There was no significant difference between LP and CON group, except that the ileal IL-10 mRNA expression was significantly higher than that of CON group. Ileal pathological scores, serum LPS levels and the expression level of ileal IL-6 mRNA were significantly lower in LP+ ANP group than those in ANP group, while the expression level of ileal IL-10 mRNA was significantly higher than that of ANP group; the expression of ileal tight junction proteins (ocludin, claudin-1, ZO-1) was significantly higher than those in ANP group, and all the differences were statistically significant (all P values <0.05). Conclusions:LP WCFS1 gavage could ameliorate the injury of pancreatic and ileal barrier in caerulein-induced ANP mice.
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Clinical features of and genetic mutations in two cases of pseudohypoparathyroidism type Ⅰ a(PHP Ⅰ a) with early-onset skin nodules were analyzed.Both of the two patients were males,and their ages at onset were 2 and 3 months respectively.They both presented with early-onset skin nodules as the main clinical manifestation,and were clinically characterized by a round face,short neck and early obesity.Histopathological examination of skin lesions showed subcutaneous ectopic osteogenesis in both patients.The first patient had low blood calcium,high blood phosphorus,high parathyroid hormone (PTH),and gene sequencing showed a heterozygous mutation c.399delT causing a T base deletion at position 399 in exon 5 of the GNAS gene.The second patient had normal blood calcium and phosphorus levels as well as normal PTH levels at early stage,and gene sequencing showed a heterozygous mutation c.939delT causing a T base deletion at position 939 in exon 9 of the GNAS gene.The blood PTH level was found to increase in the second patient after 1-year follow-up.Both the patients were confirmedly diagnosed with PHP Ⅰa.After treatment with vitamin D3,no new skin nodules occurred,and the blood calcium and phosphorus levels returned to normal.
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A case of ichthyosis follicularis,alopecia and photophobia syndrome caused by a novel mutation c.1165C＞T in the membrane-bound transcription factor protease site 2 (MBTPS2) gene was firstly reported.The proband presented with dry skin,congenital hairlessness,follicular keratotic papules,photophobia,epilepsy,and mental and motor retardation.Next-generation and Sanger sequencing analysis confirmed that the proband and his mother both had a c.1165C＞T (p.pro389Ser) mutation in exon 9 of the MBTPS2 gene.According to the clinical manifestations of the patient and genetic characteristics of the MBTPS2 gene mutation,the patient was diagnosed with ichthyosis follicularis,alopecia and photophobia syndrome.
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objective To analyze the clinical characteristics of pneumonia in patients with aplastic anemia for improving early prevention and clinical diagnosis. Methods A retrospective analysis was conducted for patients with aplastic anemia treated in our hosipital from June 2013 to April 2018. The clinical data of pneumonia were reviewed and analyzed in terms of radiological findings, pathogen distribution, and related risk factors. Results The clinical manifestations are atypical in patients with aplastic anemia. The imaging findings suggested that the infection was mainly bilateral pulmonary infection. The common pathogens were gram-negative bacteria such as Klebsiella pneumoniae. Disease type, agranulocytosis and low serum albumin level were independent risk factors for pneumonia in patients with aplastic anemia. Regular immunosuppressive therapy is a protective factor for pneumonia in patients with aplastic anemia. Conclusions The clinical manifestations of pneumonia are diverse in patients with aplastic anemia. The pathogens are mainly gram-negative bacteria. Empirical use of β-lactam-β-lactamase inhibitor combinations or quinolones is beneficial for controlling infection. Regular immunosuppressive therapy, recovery of hematopoietic function, nutritional support are effective measures to reduce the incidence of pneumonia in patients with aplastic anemia.
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Objective To analyze the clinical and biological characteristics of adult acute T-lymphocytic leukemia (T-ALL) patients carrying SET-NUP214 fusion gene,and the prognostic value of SET-NUP214 molecular marker monitoring.Methods The clinical and laboratory data of 4 adult T-ALL patients with SET-NUP214 fusion gene in the Third People's Hospital of Chengdu from January 2009 to December 2014 were analyzed retrospectively,and T cell receptor (TCR) gene rearrangement was detected to judge the differentiation and developmental stages of tumor cells in these patients.Minimal residual disease (MRD) was detected in 2 patients with follow-up specimens through detection of SET-NUP214 gene by using polymerase chain reaction.Results Four patients expressed T cell immune markers CD5,CD7,and cytoplasmic CD3 (cyCD3),and also expressed some myeloid-specific antigens.All 4 patients had the same SET-NUP214 fusion site.In the tumor cells of 4 patients,5 TCRB gene rearrangements were detected,all of which were incomplete rearrangement of DB-JB;4 patients were detected with TCRG and TCRD gene rearrangements,and all were completely rearranged.The result of MRD monitoring through SET-NUP214 fusion gene was consistent with clinical treatment outcome.Conclusions The T-ALL patients with SET-NUP214 fusion gene have some unique cell biological characteristics.SET-NUP214 fusion gene could be used as a molecular marker for MRD monitoring,and which can be used for the follow-up in the course of treatment.
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A male patient, who was aged 3 months and 12 days, presented with well-circumscribed erythema and scales on the scrotum, perineum, buttocks and perianal region at 1 month after birth. The lesions gradually involved the perioral and axillary regions, flexor aspect of the elbow, popliteal fossa and neck. Shortness of breath, crying, dysphoria and vomiting occurred without fever and cough 3 days before hospitalization. Laboratory examinations at admission showed metabolic acidosis, hyperlactacidemia, hyperammonemia and organic aciduria. Second-generation sequencing and Sanger sequencing of the holocarboxylase synthetase gene revealed a known mutation c.1522C>T in exon 9 and a novel mutation c.1796_1814del in exon 11. According to a guideline from the American College of Medical Genetics and Genomics, this novel mutation was ranked as a pathogenic mutation. The patient was diagnosed as multiple carboxylase deficiency. His clinical symptoms were improved after oral biotin treatment, no neurological symptoms or signs were observed.
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Objective To describe the study design,the characteristics of participants as well as the pedigrees included in the baseline survey of Fujian Tulou Family Cohort Study.Methods Fujian Tulou Family Cohort Study was a prospective open cohort study with a biological sample bank.A baseline survey was conducted in Tulou areas of Nanjing county in Fujian province from 2015 to 2018,including questionnaire survey,physical and biochemical indicators examinations,and blood sample collection in adults aged ≥ 18 years.In addition,family relationship of the participants was also recorded.The pedigree information of the juveniles under 18 years old were also collected.Results The baseline survey included 2 727 individuals in two clans,of whom 2 373 (87.0％) were adults,and 2 126 participants completed questionnaires,physical examinations and biochemical tests.The average age of the 2 126 participants was (57.9 ± 13.3) years,with 39.4％ being males.The current smoking rates in male and female participants were 41.2％ and 2.1％,respectively.The corresponding rates of current alcohol consumption were 19.0％ and 2.6％.For common chronic diseases,the prevalence rates were 51.3％ for hypertension,9.7％ for diabetes and 26.7％ for hyperlipemia according to the self-reported disease diagnoses,health examination results and biochemical examination results in class 1Ⅱor Ⅲ hospitals.Based on the family relationship information and genealogical data,710 pedigrees were finally identified,consisting of 5 087 family members.The numbers of five,four,three,and two generations pedigrees were 3,88,238 and 381,respectively.The pairs of the first to the fifth degree relatives were 12 039,2 662,1 511,202 and 31,respectively.Conclusion The establishment of Fujian Tulou Family Cohort provides valuable resources for exploring the genetic risk factors,environmental risk factors and gene-environment interactions contributing to the risk of common chronic diseases.
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Objective To describe the study design,the characteristics of participants as well as the pedigrees included in the baseline survey of Fujian Tulou Family Cohort Study.Methods Fujian Tulou Family Cohort Study was a prospective open cohort study with a biological sample bank.A baseline survey was conducted in Tulou areas of Nanjing county in Fujian province from 2015 to 2018,including questionnaire survey,physical and biochemical indicators examinations,and blood sample collection in adults aged ≥ 18 years.In addition,family relationship of the participants was also recorded.The pedigree information of the juveniles under 18 years old were also collected.Results The baseline survey included 2 727 individuals in two clans,of whom 2 373 (87.0％) were adults,and 2 126 participants completed questionnaires,physical examinations and biochemical tests.The average age of the 2 126 participants was (57.9 ± 13.3) years,with 39.4％ being males.The current smoking rates in male and female participants were 41.2％ and 2.1％,respectively.The corresponding rates of current alcohol consumption were 19.0％ and 2.6％.For common chronic diseases,the prevalence rates were 51.3％ for hypertension,9.7％ for diabetes and 26.7％ for hyperlipemia according to the self-reported disease diagnoses,health examination results and biochemical examination results in class 1Ⅱor Ⅲ hospitals.Based on the family relationship information and genealogical data,710 pedigrees were finally identified,consisting of 5 087 family members.The numbers of five,four,three,and two generations pedigrees were 3,88,238 and 381,respectively.The pairs of the first to the fifth degree relatives were 12 039,2 662,1 511,202 and 31,respectively.Conclusion The establishment of Fujian Tulou Family Cohort provides valuable resources for exploring the genetic risk factors,environmental risk factors and gene-environment interactions contributing to the risk of common chronic diseases.
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Objective To study expression,distribution,significance and relationship of CXCL12,CXCR4,microvessel density (MVD) in multiple myeloma (MM) bone marrow micro niche.Methods 63 cases of patients with MM were chosen as the experimental group,42 cases of healthy persons were chosen as the control group.The expression and distribution of CXCL12,CXCR4,MVD were detected by immunohistochemical method.Results Compared with the control group,the expressions of CXCL12,CXCR4,MVD in the experimental group were higher,the difference was statistically significant (P ＜ 0.05),their expressions in bone marrow were not significantly different from gender,age,immunoglobulin type and light chain classification (P＞0.05);the expression of CXCR4,MVD were positively related with CXCL12 (P＜0.05).Conclusion The expression of CXCL12,CXCR4,MVD may be related to the occurrence of MM;CXCL12/CXCR4 biological axis could promote the angiogenesis of MM bone marrow micro niche.
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Background:Acute pancreatitis can induce intestinal barrier dysfunction in its early phase,which is closely related with the progression and prognosis of the disease. Intestinal mucus layer not only serves as a physical barrier between pathogens and epithelium,but also plays a critical role in the maintenance of intestinal barrier function. Aims:To investigate the expression and role of mucin 2 (MUC2)in injured intestinal barrier in rats with acute necrotizing pancreatitis (ANP). Methods:Forty-two male Sprague-Dawley rats were randomly divided into two groups:the sham operation (SO)group and ANP group,which were induced via a retrograde injection of 3. 5% sodium taurocholate into biliopancreatic duct. Blood,pancreas and colon samples were obtained 6,12 and 24 hours after establishing the ANP model for determination of serum amylase and D-lactate (an indicator of intestinal permeability)and histopathological examination. PAS/ AB staining was used to observe the colon mucus layer and goblet cells,and the expressions of MUC2 and inflammatory cytokines in colonic tissue were detected by real-time PCR. Results:ANP models were successfully established. In ANP group,obvious colonic injury,increased intestinal permeability,thinner colon mucus layer,reduced mucin-containing goblet cells,down-regulated MUC2 mRNA and up-regulated tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β)and interferon-γ (IFN-γ)mRNAs were observed at each time point as compared with those in SO group (P < 0. 05). Spearman correlation coefficient revealed that MUC2 expression was negatively correlated with the intestinal permeability and expression of inflammatory cytokines in ANP group (P < 0. 05). Conclusions:Transcription of MUC2 is significantly down-regulated in colonic tissue of ANP rats,and might be associated with increased intestinal permeability and excessive expression of inflammatory cytokines in early phase of ANP.
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Objective To investigate the neuro-protective effect of bone marrow mesenchymal stem cells (BMSCs)transplantation in cerebral ischemia/reperfusion injury model. Methods The modified Zea-Longa suture method was adopted to establish rat middle cerebral artery occlusion(MCAO)/reperfusion injury model.Experimen-tal rats were randomly divided into blank group,model group,PBS group and BMSCs transplantation group. PBS group and BMSCs transplantation group were respectively received tail vein injection of 1 mL PBS or BMSCs(l × l06)6 h after the establishing of MCAO/reperfusion model. Neurological function was evaluated using the modified neurologic severity score(mNSS)scale at 12 h,1 d,3 d and 7 d after operation.Quantitative real-time PCR was used to detect the levels of miR-34a and miR-21 in ischemic brain tissue. Results(1)The mNSS scores in BM-SCs transplantation group showed no significant difference when compared with those in model group at 12 h and 1 d after operation(P > 0.05),whereas were significantly lower than those in model group at 3 d and 7 d(P <0.05).(2)Compared with model group,BMSCs transplantation group showed decreased level of miR-34a(P <0.01),while elevated level of miR-21 at different time points(P<0.01). Conclusions BMSCs transplantation can improve the impaired neurological function induced by cerebral ischemia/reperfusion injury,showing neuro-pro-tective effect.The mechanism may be associated with regulating the expressions of apoptosis-related microRNAs.
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Objective To determine the viability of Schistosoma japonicum cercariae by staining. Methods Schistosoma japonicum cercariae were stained by 0.4%trypan blue 0.5%methylene blue?eosin?borax M.E.B 0.5%eosin 0.5%methy?lene blue and 0.05% neutral red respectively for 5 min then they were observed under a stereoscopic microscope. Results The dead cercariae were stained in the trypan blue M.E.B eosin and neutral red but unstained in the methylene blue. The vi?tal cercariae were unstained in all the five kinds of dyes. Conclusion The staining methods by using 0.4% trypan blue 0.5%M.E.B 0.5%eosin and 0.05%neutral red can be used to determine the viability of S. japonicum cercariae.
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Objective To study the seasonal changes of glucose levels per unit soft tissue of Oncomelania hupensis. Meth-ods O. hupensis snails were collected from the beach of the Qingyi River in Wuhu City,Anhui Province from August 2012 to July 2013. They were kept in minus 80℃refrigerator immediately. The male snails were distinguished from female,and their soft tissues were collected separately after crushing their shells. The hexokinase method was used to measure the glucose concen-trations,and the results were analyzed statistically. Results The contents of glucose decreased from March to July. The lowest glucose content in the female was 1.87μg/mg in March,and that in male was 3.70μg/mg in July. Both of them increased from August and reached peak levels in September(♀=57.38μg/mg,♂=44.39μg/mg),and then gradually decreased from Octo-ber to next January and increased in February. Conclusion The contents of glucose have seasonal changes regularly in O. hu-pensis.
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ObjectiveTo investigate the effect of vasostatin on the migration of pancreatic cancer endothelial cells.Methods Ad-vasostatin with different concentrations of vasostatin was used to transfect pancreatic cancer endothelial cells.Ad-LacZ transfection and PBS was used as control.The effect of vasostatin gene mediated by adenovirus on the migration of pancreatic cancer endothelial cells was measured by woundhealing assay,transwell migration assay,and tube formation assay.ResultsThe scratched lines in PBS group and Ad-LacZ group were almost healed 48 hours later,while the lines in Ad-vasostatin group were rarely healed.At the MOI of 1,2,5,the migration rate of Ad-Laz group was ( 84.7 ± 2.6) ％,(80.7 ± 1.7 ) ％ and (81.3±4.0)％,while the corresponding values were (77.7 ±2.1)％,(67.3 ±2.1)％ and (38.8 ±2.1 ) ％ in Ad-vasostatin group.Transwell migration assay indicated that the number of migrated cells in Advasostatin group was inhibited in a dose-dependant manner,at the MOI of 5,the migration became significantly decreased (F=180.88,P ＜0.05).At the MOI of 1,5,10,the number of tubes in Ad-LacZ group was 118±6,120±6 and 82±5,while the corresponding values were 65±4,21±4 and 4 ±1 in Ad-vasostatin group.The number of tubes of pancreatic cancer endothelial cells was inhibited by Ad-vasostatin in a dose-dependant manner,at the MOI of 10, it was difficult to form the tubes (F-300.85,P＜0.05). Conclusions The vasostatin gene mediated by adenovirus has a significant inhibitory effect on the migation of pancreatic cancer endothelial cells in vitro in a dose-dependent manner.