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What is already known about this topic?: Previous research has primarily examined the issue of hepatitis B vaccine hesitancy in migrant workers and other adult populations. However, there is a lack of studies that have specifically investigated the prevalence of hepatitis B vaccine hesitancy among university students. What is added by this report?: In this study, 19.84% of students expressed hesitancy towards receiving the hepatitis B immunization. A negative correlation was observed between hepatitis B vaccine hesitancy and knowledge, attitudes, and practices related to hepatitis B. Conversely, a positive relationship was identified between hepatitis B-related knowledge and attitudes and practices. What are the implications for public health practice?: This study examines the factors contributing to vaccine hesitancy towards hepatitis B at a medical university in China. The results have significant implications for developing strategies to improve hepatitis B vaccination rates.
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A visible-light-enabled photoredox radical cascade cyclization of 2-vinyl benzimidazole derivatives is developed. This chemistry is applicable to a wide range of N-aroyl 2-vinyl benzimidazoles as acceptors, and halo compounds, including alkyl halides, acyl chlorides and sulfonyl chlorides, as radical precursors. The Langlois reagent also serves as an effective partner in this photocatalytic oxidative cascade process. This protocol provides a robust alternative for rendering highly functionalized benzo[4,5]imidazo[1,2-b]isoquinolin-11(6H)-ones.
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BACKGROUND: The incidence of chronic kidney disease among patients with diabetes mellitus (DM) remains a global concern. Long-term obesity is known to possibly influence the development of type 2 diabetes mellitus. However, no previous meta-analysis has assessed the effects of body mass index (BMI) on adverse kidney events in patients with DM. AIM: To determine the impact of BMI on adverse kidney events in patients with DM. METHODS: A systematic literature search was performed on the PubMed, ISI Web of Science, Scopus, Ovid, Google Scholar, EMBASE, and BMJ databases. We included trials with the following characteristics: (1) Type of study: Prospective, retrospective, randomized, and non-randomized in design; (2) participants: Restricted to patients with DM aged ≥ 18 years; (3) intervention: No intervention; and (4) kidney adverse events: Onset of diabetic kidney disease [estimated glomerular filtration rate (eGFR) of < 60 mL/min/1.73 m2 and/or microalbuminuria value of ≥ 30 mg/g Cr], serum creatinine increase of more than double the baseline or end-stage renal disease (eGFR < 15 mL/min/1.73 m2 or dialysis), or death. RESULTS: Overall, 11 studies involving 801 patients with DM were included. High BMI (≥ 25 kg/m2) was significantly associated with higher blood pressure (BP) [systolic BP by 0.20, 95% confidence interval (CI): 0.15-0.25, P < 0.00001; diastolic BP by 0.21 mmHg, 95%CI: 0.04-0.37, P = 0.010], serum albumin, triglycerides [standard mean difference (SMD) = 0.35, 95%CI: 0.29-0.41, P < 0.00001], low-density lipoprotein (SMD = 0.12, 95%CI: 0.04-0.20, P = 0.030), and lower high-density lipoprotein (SMD = -0.36, 95%CI: -0.51 to -0.21, P < 0.00001) in patients with DM compared with those with low BMIs (< 25 kg/m2). Our analysis showed that high BMI was associated with a higher risk ratio of adverse kidney events than low BMI (RR: 1.22, 95%CI: 1.01-1.43, P = 0.036). CONCLUSION: The present analysis suggested that high BMI was a risk factor for adverse kidney events in patients with DM.
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Short-chain dehydrogenase/reductases (SDRs) belong to the NAD(P)(H)-dependent oxidoreductase superfamily, which have various functions of catalyzing oxidation/reduction reactions and have been generally used as powerful biocatalysts in the production of pharmaceuticals. In this study, ScSDR1 and ScSDR2, two new SDRs have been identified and characterized from Stachybotrys chartarum 3.5365. Substrate scope investigation revealed that both of the enzymes possessed the ability to oxidize ß-OH to ketone specifically, and exhibited substrate promiscuity and high stereo-selectivity for efficiently catalyzing the structurally different prochiral ketones to chiral alcohols. These findings not only suggest that ScSDR1 and ScSDR2 might be potent synthetic tools in drug research and development, but also provide good examples for further engineered enzymes with higher efficiency and stereo-selectivity.
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Redutases-Desidrogenases de Cadeia Curta , Stachybotrys , Oxirredutases , Catálise , Álcoois/químicaRESUMO
The gastrointestinal tract is essential for food digestion, nutrient absorption, waste elimination, and microbial defense. Single-cell transcriptome profiling of the intestinal tract has greatly enriched our understanding of cellular diversity, functional heterogeneity, and their importance in intestinal tract development and disease. Although such profiling has been extensively conducted in humans and mice, the single-cell gene expression landscape of the pig cecum remains unexplored. Here, single-cell RNA sequencing was performed on 45â¯572 cells obtained from seven cecal samples in pigs at four different developmental stages (days (D) 30, 42, 150, and 730). Analysis revealed 12 major cell types and 38 subtypes, as well as their distinctive genes, transcription factors, and regulons, many of which were conserved in humans. An increase in the relative proportions of CD8 + T and Granzyme A (low expression) natural killer T cells (GZMA low NKT) cells and a decrease in the relative proportions of epithelial stem cells, Tregs, RHEX + T cells, and plasmacytoid dendritic cells (pDCs) were noted across the developmental stages. Moreover, the post-weaning period exhibited an up-regulation in mitochondrial genes, COX2 and ND2, as well as genes involved in immune activation in multiple cell types. Cell-cell crosstalk analysis indicated that IBP6 + fibroblasts were the main signal senders at D30, whereas IBP6 - fibroblasts assumed this role at the other stages. NKT cells established interactions with epithelial cells and IBP6 + fibroblasts in the D730 cecum through mediation of GZMA-F2RL1/F2RL2 pairs. This study provides valuable insights into cellular heterogeneity and function in the pig cecum at different development stages.
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Ceco , Intestinos , Humanos , Camundongos , Animais , Suínos , Ceco/metabolismo , Trato Gastrointestinal , Perfilação da Expressão Gênica/veterinária , Células EpiteliaisRESUMO
Objective:To investigate the efficacy and safety of thoracic duct repair in the treatment of iatrogenic thoracic duct injury in thyroid surgery.Methods:Clinical data of 2 759 cases with lymph nodes metastasis from papillary thyroid carcinomas at two tertiary referral academic medical centers, viz. Tongji Hospital of Huazhong University of Science and Technology and Zhongnan Hospital of Wuhan University were retrospectively analyzed, of which 69 cases encountered injured thoracic duct. Thirty patients received thoracic duct ligation (ligation group) before Jun. 2016, and thirty-nine with direct suture repair of lacerated duct (repair group). Age, sex, cervical clearance surgery, lymph node dissection, lymph node metastasis, whether lymph nodes were invaded extracapsular, chest catheter injection into venous points, and operation time were recorded.Results:In the thoracic duct direct suture repair group, 39 patients with a mean age of (43.2±9.3) successfully performed intraoperative thoracic duct sutures and only one patient developed CL postoperatively. In the thoracic duct ligation group, 30 ducts with a mean age of (49.5±10.0) were successfully ligated during the operation, and 5 (16.7 %) still developed CL after operations, with the highest daily drainage volume of more than 500 ml, and the median duration of chylous leakage of 3 days. The incidence of CL in the thoracic duct direct suture repair group was significantly lower than that in the ligation group (0 vs 16.7%, P=0.013), and the length of hospital stay in the repair group was also significantly shorter (6.1 (6.0,6.5) vs 10.0 (9.0,10.0) day, Z=-7.21, P=0.014) .There was no significant difference in operation time between the two groups (110.9±8.7 vs 109.3±7.9, t=0.82, P=0.421) . Conclusion:Compared with thoracic duct ligation, thoracic duct direct suture repair in patients with intraoperative thoracic duct injury may be an effective method to reduce postoperative CL.
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Objective To investigate the risk factors of postoperative pancreatitis after endoscopic retrograde cholangiopancreatography(ERCP),establish a quantitative risk prediction model,and conduct external validation.Methods The clinical data of patients with ERCP were analyzed.Among them,603 ERCP patients From January 2017 to January 2021 were selected as the modeling group,and 205 ERCP patients from March 2021 to March 2022 were selected as the validation group.Pancreatitis was diagnosed according to Atlanta standards.There were 45 cases in the modeling group and 23 cases in the validation group developed pancreatitis after ERCP.Compare the clinical data and biochemical indicators of patients with and without pancreatitis in the modeling group,and screen for risk factors of pancreatitis through multivariate Logistic regression analysis.Then,establish a risk prediction model and validate it.Results Univariate analysis showed that there were statistically significant differences in age,history of gastectomy,calculus of common bile duct,papillary foramen nodule type,pancreatic wire channel,sphincterotomy,serum total bilirubin and albumin between the two groups(P<0.05).Multivariate Logistic regression analysis showed that the history of gastrectomy(O(R) = 6.417,95%CI:1.900~21.675,P = 0.000),calculus of common bile duct(O(R) = 3.442,95%CI:1.496~7.917,P = 0.000),papillary foramen nodule type(O(R) = 2.447,95%CI:1.072~5.585,P = 0.018),pancreatic wire channel(O(R) = 3.673,95%CI:1.609~8.383,P = 0.000),sphincterotomy(O(R) = 1.758,95%CI:1.140~2.711,P = 0.004),elevated total bilirubin(O(R) = 1.415,95%CI:1.084~1.847,P = 0.008)and decreased albumin(O(R) = 1.239,95%CI:1.016~1.510,P = 0.010)were independent risk factors for post ERCP pancreatitis.Establish a risk prediction model Y =-1.023 + 1.859×(history of gastrectomy)+ 1.236×(calculus of common bile duct)+ 0.895×(papillary foramen nodule type)+ 1.301×(pancreatic wire channel)+ 0.564×(sphincterotomy)+ 0.347×(elevated total bilirubin)+ 0.214×(decreased albumin).The receiver operator characteristic curve(ROC curve)showed that the area under the curve(AUC)of the model predicting pancreatitis in the modeling and validation groups were 0.895 and 0.864,respectively.After assigning values to each variable in the model,it was divided into low risk(0~5 points),medium risk(5~10 points),and high risk(≥10 points).The actual incidence of high-risk pancreatitis in the modeling and validation groups was significantly higher than that of low-risk patients,and the actual incidence of high-risk pancreatitis was significantly higher than that of medium risk patients,with statistical significance(P<0.05).Conclusion The history of gastrectomy,calculus of common bile duct,papillary foramen nodule type,pancreatic wire channel,sphincterotomy,elevated total bilirubin and decreased albumin were independent risk factors for post ERCP pancreatitis.We have developed a quantitative risk prediction model with good predictive efficacy for pancreatitis,which has important clinical application value.
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This study aims to reveal the endogenous metabolic characteristics of acteoside in the young rat model of purinomycin aminonucleoside nephropathy(PAN) by non-targeted urine metabolomics and decipher the potential mechanism of action. Biochemical indicators in the urine of rats from each group were determined by an automatic biochemical analyzer. The potential biomarkers and related core metabolic pathways were identified by ultra-high performance liquid chromatography coupled with linear ion trap-Orbitrap mass spectrometry(UHPLC-LTQ-Orbitrap MS) combined with principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA). MetaboAnalyst 5.0 was used to establish the receiver operating characteristic(ROC) curve for evaluating the clinical diagnostic performance of core metabolites. The results showed that acteoside significantly decreased urinary protein-to-creatinine ratio in PAN young rats. A total of 17 differential metabolites were screened out by non-targeted urine metabolomics in PAN young rats and they were involved in phenylalanine metabolism and phenylalanine, tyrosine and tryptophan biosynthesis. Thirtten differential metabolites were screened by acteoside intervention in PAN young rats, and they were involved in phenylalanine metabolism and arginine and proline metabolism. Among them, leucylproline and acetophenone were the differential metabolites that were significantly recovered after acteoside treatment. These pathways suggest that acteoside treats PAN in young rats by regulating amino acid metabolism. The area under the curve of two core biomarkers, leucylproline and acetophenone, were both greater than 0.9. In summary, acteoside may restore amino acid metabolism by regulating endogenous differential metabolites in PAN young rats, which will help to clarify the mechanism of acteoside in treating chronic glomerulonephritis in children. The characteristic biomarkers screened out have a high diagnostic value for evaluating the treatment of chronic glomerulonephritis in children with acteoside.
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Humanos , Criança , Ratos , Animais , Puromicina Aminonucleosídeo , Metabolômica/métodos , Biomarcadores/urina , Cromatografia Líquida de Alta Pressão/métodos , Acetofenonas , Glomerulonefrite , Fenilalanina , AminoácidosRESUMO
This study comprehensively analyzed the active components of Sanhan Huashi Formula using qualitative and quantitative mass spectrometry techniques, laying the foundation for understanding its pharmacological substance basis. UHPLC-LTQ-Orbitrap-MS and GC-MS technologies were used to analyze and identify the volatile and non-volatile components in Sanhan Huashi Formula. UHPLC-QQQ-MS/MS technology was used to simultaneously determine the content of 27 major active components in the formula. The results showed that 308 major chemical components were identified in Sanhan Huashi Formula, among which 60 compounds were identified by comparing with reference standards, mainly including alkaloids, flavonoids, coumarins, triterpenoid saponins, amino acids, and nucleosides. GC-MS technology preliminarily identified 52 volatile compounds, with γ-eudesmol and β-eudesmol as the main components. The quantitative results demonstrated good linearity(r>0.99) for the 27 active components, indicating the stability, simplicity, and reliability of the established method. Among them, amygdalin, nodakenin, arecoline, ephedrine, and pseudoephedrine had relatively high content and were presumably the main pharmacologically active substances. In conclusion, this study systematically and comprehensively characterized the major chemical components and patterns in Sanhan Huashi Formula, providing a basis for understanding its pharmacological mechanisms and clinical applications.
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Espectrometria de Massas em Tandem , Cromatografia Líquida de Alta Pressão , Cromatografia Gasosa-Espectrometria de Massas , Reprodutibilidade dos Testes , Medicamentos de Ervas Chinesas/químicaRESUMO
Objective:To evaluate the role of interleukin-6 (IL-6) in brain tissues in cognitive impairment after myocardial infarction in mice.Methods:Forty SPF healthy male C57/BL6J mice and 40 IL-6Rα flox/flox: CAMKⅡ Cre (IL-6Rα NKO) mice, aged 9 months, weighing 34-39 g, were divided into 2 groups ( n=20 each) using a random number table method: sham operation group (Sham group, IL-6Rα NKO-Sham group) and myocardial infarction group (MI group, IL-6Rα NKO-MI group). Myocardial infarction model was developed by ligating the left anterior descending coronary artery in anesthetized animals. The same surgery was performed without ligation of blood vessels in Sham group. At 28 days after preparing the myocardial infarction model, cardiac function was assessed by cardiac doppler echocardiography, and cognitive function was assessed using the Barnes maze test. Then mice were sacrificed, and brain tissues were collected for determination of the expression of IL-6, c-Fos, ionized calcium-binding adaptor molecule 1 (Iba-1, a marker protein for microglia activation) and glial fibrillary acidic protein (GFAP) (by immunofluorescence staining) in the hippocampal CA3 region. The ultrastructure of synapses in the hippocampi was examined with a transmission electron microscope to record the number of synapses and thickness of postsynaptic density (PSD). Results:Compared with sham operation groups, the ejection fraction and shortening rate of short axis were significantly decreased, and the left ventricular end-systolic diameter was increased in myocardial infarction groups ( P<0.05). Compared with Sham group, the latency to reach the target hole was significantly prolonged in the Barnes maze test, the expression of IL-6, c-Fos, GFAP and Iba1 was up-regulated in the hippocampal CA3 region, and the number of synapses and thickness of PSD were decreased in MI group ( P<0.05). Compared with IL-6Rα NKO-Sham group, the number of synapses in hippocampal neurons was significantly decreased ( P<0.05), and no significant change was found in the other parameters in IL-6Rα NKO-MI group ( P>0.05). Compared with MI group, the latency to reach the target hole was significantly shortened in the Barnes maze test, the expression of IL-6, c-Fos, GFAP and Iba1 was down-regulated in the hippocampal CA3 region, and the number of synapses and thickness of PSD were increased in IL-6Rα NKO-MI group ( P<0.05). Conclusions:The mechanism underlying cognitive impairment after myocardial infarction may be related to hippocampal synaptic damage caused by IL-6-mediated neuroinflammatory responses in mice.
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ABSTRACT PreS/S gene mutations could impact virus secretion, infection and immune evasion. However, the relationship between PreS/S mutations and intrauterine transmission has not yet been clarified. Thus, we aimed to explore the associations between PreS/S gene mutations of HBV isolated from mothers and intrauterine transmission. We analyzed the mutations of PreS/S regions of the HBV genome in mothers with HBV DNA levels ≥ 106 IU/mL whose neonates experienced HBV intrauterine transmission (transmission group, GT) and those whose neonates did not experience intrauterine transmission (control group, GC) analyzed using clone-based sequencing. In total, 206 sequences were successfully amplified, including 98 sequences (from 21 mothers) from GT and 108 sequences (from 20 mothers) from GC of genotype C for mutational analysis. Among the 1203 nucleotides of PreS/S regions, there were 219 (18.20%) base substitutions, of which 103 (47.03%) base mutations caused amino acid changes. F80S, A90V and I68T were mutation hotspots. Mothers in GT had a higher mutation rate of A90V in the PreS1 gene than mothers in GC. The A90V mutation increased the risk of HBV intrauterine transmission after adjusting the maternal age and the mode of delivery (OR = 6.23, 95% CI: 1.18-32.97). Moreover, the area under the ROC curve (AUC) for intrauterine transmission due to A90V and a combination of A90V with the mode of delivery were 0.723 (95% CI: 0.575 to 0.891, P = 0.011) and 0.848 (95% CI: 0.723 to 0.972, P < 0.001), respectively. Mothers with the A90V mutation in the PreS1 gene may be a potential risk factor for HBV intrauterine transmission.
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ObjectiveIn order to establish a systematic quality evaluation system for Fritillariae Cirrhosae Bulbus adulteration, portable near-infrared (NIR) spectroscopy was used to identify Fritillariae Cirrhosae Bulbus and its adulterants and detect their adulteration quantity. MethodA total of 72 batches of Fritillariae Cirrhosae Bulbus samples were collected and 570 batches of adulterated products (dry bulbs of Fritillaria thunbergii, F. ussuriensis, F. pallidiflora and F. hupehensis, Bulbus Tulipae, flour) were prepared, NIR spectral data of samples were collected by the portable NIR spectrometer. Linear discriminant analysis (LDA) was used to establish the qualitative correction models of Fritillariae Cirrhosae Bulbus-adulterants and adulterants of different categories, partial least squares (PLS) was used to establish the quantitative correction models of adulteration quantity of different kinds of adulterants. ResultThe recognition rates of qualitative analysis model of Fritillariae Cirrhosae Bulbus and its adulterants were 99.49% (calibration set) and 100.00% (validation set), respectively. In different adulterant models, the recognition rates of calibration set and validation set were 70.47% and 73.68%, respectively. Moreover, the correlation coefficients of validation set (R2P) of the six quantitative models of adulteration ratio were 0.840 2 (Fritillariae Cirrhosae Bulbus adulterated with F. thunbergii dry bulbs), 0.960 2 (Fritillariae Cirrhosae Bulbus adulterated with F. ussuriensis dry bulbs), 0.765 7 (Fritillariae Cirrhosae Bulbus adulterated with F. pallidiflora dry bulbs), 0.902 5 (Fritillariae Cirrhosae Bulbus adulterated with F. hupehensis dry bulbs), 0.957 4 (Fritillariae Cirrhosae Bulbus adulterated with Bulbus Tulipae), 0.976 1 (Fritillariae Cirrhosae Bulbus adulterated with flour), the root mean square error of prediction (RMSEP) were 10.948 5, 5.463 9, 13.256 4, 8.549 2, 5.655 3, 4.235 6, respectively. The two qualitative models and six quantitative models showed good prediction performance. ConclusionThe portable NIR spectroscopy can be used to identify Fritillariae Cirrhosae Bulbus and its adulterants in real time, the method is rapid and accurate, which can meet the requirements of nondestructive identification of Fritillariae Cirrhosae Bulbus on site.
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Objective: To observe the efficacy of long-retaining scalp acupuncture plus interactive training in improving upper- extremity dysfunction in cerebral stroke patients. Methods: Ninety-five patients with upper-extremity dysfunction after cerebral stroke were randomized into two groups, with 48 cases in the treatment group and 47 cases in the control group. Conventional internal medicine treatment was offered to both groups. In both groups, Anterior Oblique Line of Vertex-temporal (MS 6, the middle 2/5) and Posterior Oblique Line of Vertex-temporal (MS 7, the middle 2/5) were selected from the same side of the brain lesion (the side apposing to the hemiplegic limb) for scalp acupuncture treatment. In the treatment group, the scalp acupuncture needles were retained for 7 h, in combination with interactive training, while the needles were also retained for 7 h in the control group but without interactive training. Prior to treatment and at 2-week and 4-week treatment, the two groups were scored using the functional test for the hemiplegic upper extremity-Hong Kong (FTHUE-HK) and simplified Fugl-Meyer assessment-upper extremity (FMA-UE). Results: The total effective rate was 97.9% in the treatment group, higher than 74.5% in the control group (P<0.01). The FTHUE-HK score was higher at 2-week and 4-week treatment than before treatment in both groups, presenting statistically significant intra-group differences (all P<0.001); the FTHUE-HK score was higher at 4-week treatment than at 2-week treatment in both groups, presenting statistically significant intra-group differences (both P<0.001). At 2-week and 4-week treatment, the FTHUE-HK score was higher in the treatment group than in the control group, showing significant between-group differences (both P<0.05). During the whole treatment process, the treatment group had higher FTHUE-HK scores compared with the control group, but there was no statistical significance comparing the change of the score between the two groups at 2-week treatment (P>0.05), while the between-group difference in the change of the score was statistically significant at 4-week treatment (P<0.05). The FMA-UE score was higher at 2-week and 4-weeks treatment than before treatment in both groups, presenting statistically significant intra-group differences (all P<0.001); the FMA-UE score was higher at 4-week treatment than at 2-week treatment in both groups, presenting statistically significant intra-group differences (both P<0.001). At 2-week and 4-week treatment, the FMA-UE was higher in the treatment group than in the control group, and the between-group differences were statistically significant (both P<0.01). The FMA-UE score rose gradually with the increase of treatment session, and there was statistical significance comparing the change of the score between the two groups at 2-week and 4-week treatment, respectively (both P<0.05). Conclusion: Long-retaining scalp acupuncture plus interactive training results in more significant efficacy than long-retaining scalp acupuncture alone in improving the upper-limb dysfunction after cerebral stroke and the advantage becomes more notable after 2-week consecutive treatment.
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Objective To investigate status of helicobacter pylori (Hp) infection in Nantong Area and its related factors. Methods A cluster of 1 680 people who underwent a physical examination at a hospital in the Nantong area from June 2020 to February 2021 were selected as the subjects of this survey. All subjects were tested for Hp infection through the 14C-urea breath test and the Hp infection status analyzed. At the same time, the self-filled and interrogative questionnaires made by our hospital were used to collect general data of the research subjects, and then single-factor and multi-factor logistic regression was used to analyze the related factors affecting Hp infection in the physical examination population in Nantong area. Results Among the 1 680 medical examiners investigated in this study, 980 were positive for Hp infection, and the overall positive rate of infection was 58.33%. The results of univariate analysis found that Hp infection was related to the age, drinking, hotness, frequent eating of pickled food, family history of gastric cancer, and chronic gastritis in the physical examination population in Nantong area (P60 years old (OR=3.46, 95%CI: 1.35-8.84), drinking (OR=5.22, 95%CI: 2.79-9.77), addiction to spicy food (OR=5.47, 95%CI: 2.45-9.30), regular consumption of pickled foods (OR=5.28, 95%CI: 3.48-7.92), family history of gastric cancer (OR=4.29, 95%CI: 2.86-76.45), chronic gastritis (OR=5.07, 95%CI: 2.79-9.22) were all related factors affecting Hp infection(P<0.05). Conclusion Positive rate of HP infection in physical examination population in Nantong area is high, and it is related to age, drinking alcohol, spicy food, eating pickled food frequently, family history of gastric cancer, chronic gastritis and other factors. To strengthen the publicity and education of HP infection, establish good eating habits and reduce the risk factors to prevent HP infection in physical examination population in Nantong area.
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In the research and development of new drugs, it is very important to investigate the in vitro metabolism of candidate drugs. Traditional models such as liver microsomes have many limitations, while the in vitro model of recombinant human drug metabolizing enzymes is considered as an important and useful approach because of its convenient access, stable activity and low cost. In this study, six major human UDP-glucuronosyltransferases (UGTs) genes (UGT1A1, 1A3, 1A4, 1A6, 1A9 and 2B7) were cloned from human liver cDNA and heterologously expressed in Saccharomyces cerevisiae and baculovirus-infected insect cell. UGT1A1, 1A3, 1A6 and 1A9 were successfully expressed in yeast and showed glucuronidation activity against a variety of different structural types of substrates, but their activities were low. All six UGTs were successfully expressed and exhibited significantly improved glucuronidation activity when Trichopolusia ni cells BTI-TN5B1-4 (High Five) were used as the host. The recombinant human UGTs expressed in insect cells can catalyze the glucuronidation of their specific substrates, and the glucuronidation products were synthesized at milligram-scale with yields of 13%-66% for the first time, of which the structures were identified via MS, 1H NMR, and 13C NMR spectroscopic analysis. Above all, the recombinant human UGTs yeast and insect cell expression systems constructed in this study can be used for in vitro metabolism evaluation in the early stage of new drugs research and development, and also provide a new tool for the synthesis of glucuronide metabolites.
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Objective:To investigate the expression of estrogen receptor alpha (ERα) protein and BRAF V600E protein in thyroid papillary carcinoma (PTC) and their relationship with clinical factors of PTC. Methods:The expression of ERα and BRAF V600E protein in 1 105 PTC patients was detected by immunohistochemistry. The relationship among ERα, BRAF V600E protein and clinical factors were analyzed. Results:Positive ERα protein was correlated with maleness (χ 2= 6.087, P=0.001), age< 45 years old (χ 2=5.197, P=0.023) and multifocal tumors (χ 2=4.446, P=0.035). Positive BRAF V600E protein was correlated with positive ERα protein (χ 2=6.209, P=0.013), Hashimoto thyroiditis (χ 2=29.388, P<0.001), no lateral lymph node metastasis (χ 2=6.849, P=0.009) and multifocal tumors (χ 2=9.596, P=0.035). Conclusions:ERα expression is more common in male patients, patients younger than 45 years of age, those with multifocal tumors and positive BRAF V600E protein. BRAF V600E protein may inhibit Hashimoto's thyroiditis, tumor growth and the occurrence of lateral lymph node metastasis, and promote the occurrence of multiple focal tumors.
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Background and AIM: To explore the efficacy treatment regimen in refractory PBC. METHODS: Triple treatment including ursodeoxycholic acid, prednisolone and immunosuppressant was prescribed to 47 refractory patients. Biochemistries, immune parameters, non-invasive liver fibrosis assessments were measured during follow-up. RESULTS: Triple therapy resulted in significant decrease in ALP, GGT, ALT, AST, TBIL, ALB, IgG, IgM, APRI, FIB-4 and S-INDEX. The biochemical cumulative normalization rates of ALP and other biochemical parameters were higher in long-term follow-up. Poor outcome was observed in patients with lower ALB, higher TBIL, PT, sp100 positivity and advanced liver pathology at baseline. Osteoporosis and bone fracture were observed in 15% patients. CONCLUSIONS: Triple therapy is associated with marked decrease and normalization of ALP and other parameters. ALB, TBIL, PT, sp100 and pathology were related with poor outcome. Osteoporosis should be closely monitored
ANTECEDENTES Y OBJETIVO: Explorar el régimen de tratamiento de eficacia en PBC refractario. MÉTODOS: Se prescribió un tratamiento triple que incluyó ácido ursodesoxicólico, prednisolona e inmunosupresor a 47 pacientes refractarios. Las bioquímicas, los parámetros inmunes, las evaluaciones no invasivas de fibrosis hepática se midieron durante el seguimiento. RESULTADOS: La triple terapia resultó en una disminución significativa de ALP, GGT, ALT, AST, TBIL, ALB, IgG, IgM, APRI, FIB-4 y S-INDEX. Las tasas de normalización bioquímica acumulada de ALP y otros parámetros bioquímicos fueron mayores en el seguimiento a largo plazo. Se observó un resultado deficiente en pacientes con ALB más bajo, TBIL más alto, PT, positivo de SP100 y enfermedad hepática avanzada al inicio del estudio. La osteoporosis y la fractura ósea se observaron en el 15% de los pacientes. CONCLUSIONES: La triple terapia se asocia con una marcada disminución y normalización de ALP y otros parámetros. ALB, TBIL, PT, SP100 y la enfermedad se relacionaron con un mal resultado. La osteoporosis debe estar bajo estrecha supervisión
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Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Ácido Ursodesoxicólico/uso terapêutico , Prednisolona/uso terapêutico , Colangite/tratamento farmacológico , Cirrose Hepática Biliar/tratamento farmacológico , Resultado do Tratamento , Estudos de Coortes , Imunossupressores/uso terapêutico , Estudos Retrospectivos , BiópsiaRESUMO
AIM:To establish a method for obtaining specific cells in solid tumor tissue by sorting of CD11b + myeloid cells in hepatic metastases from colorectal cancer.METHODS:Tumor tissues were prepared into single cell suspension by mechanical method combined with enzyme digestion,and then the CD11 b + myeloid cells were isolated by flow cytometry.The sorted cells were identified by immunocytochemistry.The viability and morphologiy of the sorted cells were evaluated by Giemsa and Typan blue staining.The cell purity was evaluated by flow cytometry.RESULTS:Sufficient numbers of CD11b+cells with high purity were isolated by sorting with flow cytometry from the single cell suspension prepared by mechanical and enzyme digestion.The purity of the cells was confirmed by statistical analysis (P < 0.05).The positive rates of the cells before and after sorting were significantly different (P <0.01).The positive cells were verified by immunocytochemical method.Meanwhile,the sorted cells had complete morphology and good activity.CONCLUSION:The CD11b + myeloid cells in solid tumor tissue can be isolated by flow cytometry from the machine-enzyme digestion suspension with high purity,good activity and complete morphology.
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AIM:To establish a method for obtaining specific cells in solid tumor tissue by sorting of CD11b + myeloid cells in hepatic metastases from colorectal cancer.METHODS:Tumor tissues were prepared into single cell suspension by mechanical method combined with enzyme digestion,and then the CD11 b + myeloid cells were isolated by flow cytometry.The sorted cells were identified by immunocytochemistry.The viability and morphologiy of the sorted cells were evaluated by Giemsa and Typan blue staining.The cell purity was evaluated by flow cytometry.RESULTS:Sufficient numbers of CD11b+cells with high purity were isolated by sorting with flow cytometry from the single cell suspension prepared by mechanical and enzyme digestion.The purity of the cells was confirmed by statistical analysis (P < 0.05).The positive rates of the cells before and after sorting were significantly different (P <0.01).The positive cells were verified by immunocytochemical method.Meanwhile,the sorted cells had complete morphology and good activity.CONCLUSION:The CD11b + myeloid cells in solid tumor tissue can be isolated by flow cytometry from the machine-enzyme digestion suspension with high purity,good activity and complete morphology.