[Association between anti-tissue transglutaminase antibody titers and duodenal histopathology among adults with celiac disease].

To evaluate the association between serum anti-tissue transglutaminase antibody (anti-tTG) titers and the severity of histological damage to the duodenal mucosa and to predict a possible anti-tTG cutoff value for diagnosing celiac disease (CD) and villous atrophy in the domestic population. Clinical and pathological data from 76 adult CD patients with positive anti-tTG titers and duodenal biopsy results who were treated at the People's Hospital of Xinjiang Uygur Autonomous Region from July 2017 to January 2022 were retrospectively analyzed. The correlation between anti-tTG titers and the severity of duodenal mucosal damage was statistically assessed to predict the optimal anti-tTG titer cut-off value for diagnosing CD and villous atrophy. Of the 76 patients, 10 had underlying CD, and of the 66 patients with duodenal histopathology, four were Marsh Ⅰ, six were Marsh Ⅱ, and 56 were Marsh Ⅲa-c grade. In adults with CD, anti-tTG titers were shown to be associated with the severity of histological damage to the duodenal mucosa. When the anti-tTG level was ≥5 times the upper limit of normal (ULN), the sensitivity and specificity for diagnosing CD were 83.9% and 92.9%, respectively. When the anti-tTG titer was ≥8 times the ULN, the sensitivity and specificity for diagnosing villous atrophy were 67.9% and 90.0%, respectively. Anti-tTG levels had a strong predictive value for diagnosing CD in adults when titers exceeded 10 times the ULN. Thus, the anti-tTG cut-off value can be combined with clinical judgment to diagnose CD, limiting the use of invasive endoscopy.

[Imaging study on the safety of axial pedicle screw placement by the position of the screw trajectory tip on the anteroposterior and lateral radiographs].

Objective: To propose a method to judge the safety of axial pedicle screw placement based on the position of the tip of the screw trajectory on the anteroposterior and lateral X-ray radiographs. Methods: The cervical CT data of 40 patients admitted to the Second Affiliated Hospital of Wenzhou Medical University from December 2020 to December 2021 were selected, including 24 males and 16 females, with a mean age of (47.6±13.2) years. Based on the three-dimensional model reconstruction of Mimics software and its function of X-ray, the transmission of the axial pedicle screw and its anteroposterior and lateral films was simulated. The position of the tip of the simulated screw trajectory was divided into 5 regions (regions Ⅰ-Ⅴ) from the inside to the outside on the anteroposterior virtual radiographs, and the upper and lower regions (regions a, b) on the lateral virtual radiographs. By adjusting the direction of the screw, the tip of the screw was located in the corresponding 10 regions (80 screws in each area) on the virtual projections of the anteroposterior and lateral virtual radiographs respectively, and its accuracy was analyzed by CT to determine whether each screw penetrated the medial wall of the pedicle or vertebral artery foramen. The anteroposterior and lateral X-rays and postoperative CT data of 34 patients who underwent axial pedicle screw placement (67 axial pedicle screws were placed in total) from January 2014 to December 2021 were collected, including 18 males and 16 females, with a mean age of (45.8±14.1) years. The position of the tip of the screw trajectory on the anteroposterior and lateral films was divided in the same way. The number of screws in the corresponding 10 positions was counted, and CT analysis was used to determine whether each screw penetrated the medial wall of the axial pedicle or the vertebral artery foreman. Results: The results of the imaging simulation screw placement study showed that the perforation rate of the vertebral artery foramen in region Ⅳ and Ⅴ was 75.0% (120/160) and 100% (160/160), respectively, while the perforation rate of the medial wall of the axial pedicle in the region Ⅰ was 85.6%(137/160). The failure rate in regions Ⅱ and Ⅲ was relatively lower, and the performance of simulated screws located in the region a was better than those in region b. The perforation rates of the medial wall in regions (a-Ⅱ) and (a-Ⅲ) was 7.5% (6/80) and 0 (0/80), respectively, and the perforation rates of the vertebral foramen was 0 (0/80) and 21.3% (17/80), respectively. The retrospective imaging study also showed a higher rate of placement failure in regions Ⅰ, Ⅳ and Ⅴ, and relatively lower in regions Ⅱ and Ⅲ. There were total of 15 screws in region a-Ⅱ and a-Ⅲ, and no destruction of the medial wall of the axial pedicle and the vertebral artery foreman occurred there. Conclusions: Regions a-Ⅱ and a-Ⅲ are the "safety areas" of the tip of the pedicle screw trajectory in the axial vertebra. By analyzing the tip of the pedicle screw trajectory on the anteroposterior and lateral radiographs, the operator can determine the reasonable trajectory of axial pedicle screw placement, prevent the injury of the cervical spinal cord and vertebral artery, and reduce the risk of operation.

[Clinical and microbiological characteristics of hypervirulent Klebsiella pneumoniae lung abscess].

Objective: To study the clinical and microbiological characteristics of hypervirulent Klebsiella pneumoniae (hvKP) lung abscess, and to compare with the classic Klebsiella pneumoniae (cKP) lung abscess. Methods: A total of 18 patients with Klebsiella pneumoniae lung abscesses admitted to the First Affiliated Hospital of Zhejiang University School of Medicine from January 2017 to September 2020 enrolled. The strains with positive result of string test were defined as hvKP, and the negative strains were defined as cKP. The patients' basic diseases, symptoms, laboratory data and other clinical characteristics were collected. The microbiological characteristics of the strains included as following: VITEK method to determine the in vitro susceptibility of the strains to antibiotics; PCR to detect the capsular serotypes and virulence genes. The differences in clinical characteristics and microbiological characteristics of strains between hvKP group and cKP group were compared. Results: Among the 18 patients with Klebsiella pneumoniae lung abscess, 12 were hvKP infection, mainly male (10 cases), with a median age of 59.0 years; 8 cases in the hvKP group had an onset time of ≤2 weeks, and the median onset time was 10.5 days. There were significantly more diabetes (12 cases) and extrapulmonary abscesses (11 cases) in hvKP group than those in cKP group (both P<0.001). The extrapulmonary abscesses in the hvKP group were mainly liver abscesses (10 cases), and 4 cases were multi-site (≥3) abscesses. The number of indwelling catheters and invasive procedures before infection were higher in cKP group than those in hvKP group (both P=0.025). The imaging of Klebsiella pneumoniae lung abscess was mainly subpleural with the size of less than 10 cm. There were more multiple abscesses cases in hvKP group (9 cases) than cKP group (P=0.009). The median interval time between the detection of a pulmonary abscess and an extrapulmonary abscess was 1.0 day. The resistance rate of common antibiotics was significantly lower in hvKP than cKP. Conclusions: hvKP lung abscesses are more common in patients with diabetes, and the clinical manifestations are nonspecific. The lung imaging manifestations are multiple subpleural abscesses, indicating hematogenous dissemination. Liver abscesses were present in most cases, suggesting the source of infection. The main virulent gene of hypervirulent Klebsiella pneumoniae is aero. For patients with hvKP lung abscess, attention should be paid to finding hidden lesions.

[Comparative treatment analysis of upper gastroenterology submucosal tumors originating from muscularis propria layer: submucosal tunneling endoscopic resection versus endoscopic submucosal excavation].

OBJECTIVE: To evaluate the efficacy and safety of submucosal tunneling endoscopic resection (STER) and endoscopic submucosal excavation (ESE) for upper gastroenterology submucosal tumors (SMT) originating from the muscularis propria (MP) layer. METHODS: Clinicopathological and endoscopic data of 42 cases with upper gastroenterology tumors originating from the MP layer who were treated with STER (n=28) or ESE (n=14) between April 2013 and December 2016 in Peking University Third Hospital were retrospectively analyzed. The treatment and complications of the two groups were compared. RESULTS: In the study, 42 cases were all resected by therapeutic endoscopy successfully．There was no significant difference (STER vs. ESE) in gender, age, mean tumor size [1.5 (1.0-6.0) cm vs. 1.3 (0.5-2.0) cm, P=0.056]. STER was superior to ESE with reduced sutured time [3.5 (1.0-11.0) min vs. 8.0 (2.0-33.0) min, P=0.006], but more resection time [46.5 (11.0-163.0) min vs.19.5 (6.0-56.0) min, P=0.007]. There was statistical difference between the two groups in resection time or sutured time, but no significant difference (STER vs. ESE) in total operative time [52.0 (14.0-167.0) min vs. 31.5 (10.0-88.0) min, P=0.080]. En bloc resection rates (92.9% vs. 85.7%), hospital stay duration and complications (10.7 vs. 0.0) were similar in the STER and ESE groups. One case developed mediastinal emphysema and 2 pneumonia after operation in STER group, and all of them recovered uneventfully after conservative treatments; There were no complications in the ESE group. After operation, 28 cases of leiomyoma and 14 cases of stromal tumor were diagnosed by routine pathological and immunohistochemical staining. Among them, 6 cases of stromal tumors in group STER were all extremely low risk, 4 cases of stromal tumors in group ESE were extremely low risk, 4 cases of stromal tumors in group ESE were medium risk (the size of the lesion was about 1.0-2.0 cm, and mitotic figures counted (6-8)/50 high power field). The median follow-up time of all the patients was 46.5 (24-60) months, and the shortest follow-up time for medium risk stromal tumors was 32 months. No residual tumor, recurrence and implantation in the tunnel were observed. CONCLUSION: STER or ESE can be used as an effective and safe option for treatment of submucosal tumors originating from the muscularis propria of the upper digestive tract. Compared with STER, ESE had shorter resection time but longer wound closure time. There was no significant difference in total operation time.

Antrodia cinnamomea reduces obesity and modulates the gut microbiota in high-fat diet-fed mice.

BACKGROUND: Obesity is associated with gut microbiota dysbiosis, disrupted intestinal barrier and chronic inflammation. Given the high and increasing prevalence of obesity worldwide, anti-obesity treatments that are safe, effective and widely available would be beneficial. We examined whether the medicinal mushroom Antrodia cinnamomea may reduce obesity in mice fed with a high-fat diet (HFD). METHODS: Male C57BL/6J mice were fed a HFD for 8 weeks to induce obesity and chronic inflammation. The mice were treated with a water extract of A. cinnamomea (WEAC), and body weight, fat accumulation, inflammation markers, insulin sensitivity and the gut microbiota were monitored. RESULTS: After 8 weeks, the mean body weight of HFD-fed mice was 39.8±1.2 g compared with 35.8±1.3 g for the HFD+1% WEAC group, corresponding to a reduction of 4 g or 10% of body weight (P<0.0001). WEAC supplementation reduced fat accumulation and serum triglycerides in a statistically significant manner in HFD-fed mice. WEAC also reversed the effects of HFD on inflammation markers (interleukin-1ß, interleukin-6, tumor necrosis factor-α), insulin resistance and adipokine production (leptin and adiponectin). Notably, WEAC increased the expression of intestinal tight junctions (zonula occludens-1 and occludin) and antimicrobial proteins (Reg3g and lysozyme C) in the small intestine, leading to reduced blood endotoxemia. Finally, WEAC modulated the composition of the gut microbiota, reducing the Firmicutes/Bacteroidetes ratio and increasing the level of Akkermansia muciniphila and other bacterial species associated with anti-inflammatory properties. CONCLUSIONS: Supplementation with A. cinnamomea produces anti-obesogenic, anti-inflammatory and antidiabetic effects in HFD-fed mice by maintaining intestinal integrity and modulating the gut microbiota.

[Expression and distribution of programmed death receptor 1 and T cell immunoglobulin mucin 3 in breast cancer microenvironment and its relationship with clinicopathological features].

Objective: To explore the expression and distribution of programmed death receptor 1 (PD-1) and T-cell immunoglobulin mucin 3 (TIM-3) in breast cancer microenvironment and analyze the their correlation with the clinicopathological features. Methods: The specimens of tumor tissue and adjacent tissues from 30 patients with infiltrative breast cancer who were diagnosed as breast cancer from June 2016 to May 2017 in The First Hospital of Jiaxing were collected, and the specimen were divided into two parts along the center. After embedding and cryosectioning, the expression and distribution of PD-1 and TIM-3 protein in tumor tissues were observed by immunofluorescence staining. Another part of the specimen was cut and digested, and non-continuous density gradient centrifugation was used to extract tumor-infiltrating lymphocytes (TILs), real-time quantitative PCR (qRT-PCR) was used to detect the mRNA expression of PD-1 and TIM-3 in TILs. Meanwhile, the protein expression was determined by Western blotting. The relationship between the expression of PD-1 and TIM-3 and pathological parameters of breast cancer was analyzed with correlation analysis. Results: Immunofluorescence results showed that more PD-1 and TIM-3 positive cells were observed in the tumor tissues compared with the tumor-adjacent tissues. The qRT-PCR showed that the expression of PD-1 and TIM-3 mRNA in TILs were both significantly higher than those in paracancerous tissues (3.09±0.38 vs 1.26±0.23, 3.42±0.31 vs 1.57±0.29, t=4.16, 4.37, both P<0.05). At the protein level, the expression of PD-1 and TIM-3 in tumor tissue lymphocytes(0.66±0.08, 0.80±0.11) was significantly higher than those in cancerous tissues(0.10±0.01, 0.26±0.02) (t=6.79, 4.57, both P<0.05). There were significant differences in the expression of PD-1, TIM-3 mRNA in the TILs between the different tumor histological grades, tumor sizes, lymph node metastasis (t=2.22-2.99, all P<0.05). Correlation analysis showed that there was a significant positive correlation between the expression of PD-1 and TIM-3 in tumor tissues (r=0.616, P<0.01). Conclusions: In the breast cancer microenvironment, PD-1, TIM-3-mediated signaling pathway plays an important role in the occurrence and development of breast cancer, it provides a new basis for the combination therapy of breast cancer.

[Value of MRI in the pre-operative diagnosis and classification of oblique vaginal septum syndrome].

Objective: To explore the role of MRI in the pre-operative diagnosis and classification of oblique vaginal septum syndrome (OVSS) . Methods: A retrospective analysis of the clinical records and pre-operative MRI images of 19 patients with surgery proved OVSS was carried out. Two experienced radiologists reviewed the pre-operative pelvic MRI of the 19 patients in consensus blind to the surgery results. Characteristics including malformations of the uterus, cervix and vagina, the diagnosis of the disorder and classification were evaluated. Pre-operative MRI diagnosis and classification were correlated with surgical findings. Results: Mean age of onset of symptoms for the 19 patients was 15 years (ranged 9-25 years) , and mean age of menarche was 12 years. Ten patients suffered from dysmenorrhea or lower abdominal pain, 5 patients complained of vaginal discharge, 3 patients had a history of irregular menstruation, 1 patient suffered from primary infertility. All 19 patients showed uteri didelphys. Eighteen patients showed vaginal oblique septum.One patient showed cervical atresia.MRI was completely correlated with the surgery in the pre-operative diagnosis of OVSS. MRI classification was in line with surgery in 17 patients, including 9 patients with imperforate septum (typeⅠ) , 6 patients with perforate septum (type Ⅱ) , 1 patient with imperforate septum and cervical fistula (type Ⅲ) , and another one with cervical agenesis (type Ⅳ) . One case of type Ⅱ was misdiagnosed as type Ⅰ, another one of type Ⅰ was misdiagnosed as type Ⅲ. Pre-operative MRI classification was correlated with surgery in 17 out of 19 patients. Conclusion: Pre-operative MRI allows excellent manifestation and accurate diagnosis of OVSS, and could also facilitate the evaluation of the classification.

[Efficacy and outcome of transcatheter closure of patent foramen ovale in patients with cryptogenic stroke].

Objective: To investigate the efficacy and outcome of transcatheter patent foramen ovale (PFO) closure in patients with cryptogenic stroke (CS). Methods: Sixty consecutive patients with cryptogenic stroke who undertook transcatheter PFO closure between May 2015 and September 2017 in Beijing Tiantan Hospital were enrolled in this prospective study.Transcranial Doppler (TCD) bubble test was performed and right-left shunt(RLS) was confirmed in all patients.Closure success rate,effective closure rate, complications, recurrence of ischemic stroke and new onset atrial fibrillation were evaluated. Results: A total of 60 patients (42 male,age range 24-68 (47±11)years) were included in the study.PFO size (motionless state) was (1.6±0.6)mm.RLS before closure was graded and 11 patients had moderate RLS and 48 patients had large RLS (include 41 patients who experienced shower or curtain effect).Closure success rate was 100% (60/60).No severe complications were observed.At 6 months,45 patients completed TCD bubble test.Of these, 4 patients suffered from moderate to large residual and thus effective closure rate was 91%(41/45).The mean follow-up period was 2-29 (median 12) months. During the follow-up, only 1 patient experienced recurrent cerebral infarction.New onset atrial fibrillation was not detected. Conclusion: Transcatheter PFO closure is effective,safe and related with a good outcome in reduction of recurrent CS for patients with PFO.

Efficacy and safety of three adefovir-based combination therapies in HBeAg-positive chronic hepatitis B patients with suboptimal response to adefovir monotherapy.

Although high potent nucleos(t)ide analogues are strongly recommended as first-line therapy for chronic hepatitis B (CHB) in China, some patients are still being treated with adefovir disoproxil (ADV), especially those low-income patients whose health insurance could not reimburse the drug cost. Therefore, the management of patients who have failed ADV therapy or who sustained renal damage during ADV therapy remains an important clinical problem in China. This retrospective study aimed to compare the efficacy and safety of lamivudine (LAM), telbivudine (LdT) or entecavir (ETV) add-on strategies to optimize the treatment of patients with prior suboptimal response to ADV monotherapy. A total of 277 eligible patients were included in this study, and the baseline characteristics were similar among the LAM + ADV (n = 116), LdT + ADV (n = 72) and ETV + ADV (n = 89) groups. At week 96, both the proportion of undetectable HBV DNA (81.03% for LAM + ADV, 84.72% for LdT + ADV and 88.76% for ETV + ADV; P = .317) and ALT elevation (5.17% for LAM + ADV, 4.17% for LdT + ADV and 4.49% for ETV + ADV; P = 1.000) were similar among the three groups; also, a significant decline in liver stiffness was observed in each group from baseline to week 96. At week 96, the rate of HBeAg seroconversion was significantly higher in LdT + ADV than in LAM + ADV (26.39% vs 13.79%, P = .031) and ETV + ADV (26.39% vs 10.11%, P = .007). During the 96 weeks, no obvious renal injury was reported in any of the three groups, but an improvement in eGFR was found in LdT + ADV compared with LAM + ADV and ETV + ADV. In summary, all three combination strategies provide good control of virus replication, but the LdT + ADV combination therapy may yield better HBeAg seroconversion and eGFR improvement.

[Correlation analysis of the pre-pregnant body mass index, the gestational weight gain and umbilical cord blood C peptide].

Objective: To explore the correlation among the pre-pregnant body mass index (BMI), gestational weight gain (GWG) and umbilical cord blood C peptide, and to investigate the influence of maternal weight management on the incidence of baby long-term metabolic syndrome. Methods: During May to Aug.2015, 485 pregnant women in Zhejiang Taizhou first people's hospital and Taizhou Huangyan maternal &child care service centre were selected in random and divided into four groups according to pre-pregnant BMI: low body mass, normal body mass, over body mass and obese group, and also divided into two groups for getting gestational diabetes mellitus (GDM) or not. According to the gestational weight gain (GWG), all the cases were divided into two groups: above the Institute Of Medicine (IMO) 2009 recommendations or not. According to the outcome, the GDM group which had received weight control treatment, was divided into successful treat group or not. At last, we tested the umbilical cord blood C peptide and birth weight of each newborn and compared the difference in all subgroups. The correlation between the umbilical cord blood C peptide and birth weight were analysed. Results: (1) In the pre-pregnant BMI groups, there were significant differences of incidence of GDM (P<0.05). But, there were no significant differences of incidence of GWG in four groups (P>0.05). (2) Newborn birth weight and the umbilical cord blood C peptide were positively correlated (r=0.673, P<0.05). (3) Umbilical cord blood C peptide showed the correlation in all subgroups for different pre-pregnant BMI, GDM or not, overweight or not (P<0.05), but only the GDM or not group had relation to newborn birth weight in all subgroups. (4) All groups showed correlations with neonatal umbilical cord blood C peptide, the GDM mostly (ß=0.58), pre-pregnant BMI secondly (ß: 0.36, 0.38) , and GWG weakly (ß=0.17). (5) By stratification analysis, in GDM group, low body mass was negatively correlated with umbilical cord blood C peptide (P<0.05, ß=-1.41); in non-GDM group, over body mass group and obsess group were positively correlated with umbilical cord blood C peptide (P<0.05, ß=0.37, 0.46). Conclusion: There was direct correlation between the pre-pregnant body mass index, the gestational weight gain and umbilical cord blood C peptide. Suitable maternal weight control especially pre-pregnant body mass index control will lower the baby long-term metabolic syndrome incidence.

[Effects and mechanisms of ursodeoxycholic acid on isoprenaline-Induced myocardial fibrosis in mice].

Objective: To investigate the effects and possible mechanisms of ursodeoxycholic acid (UDCA) on myocardial fibrosis in mice. Method: To observe the expression of transforming growth factor(TGF) -ß1, CTGF, MMPs and the degree of myocardial fibrosis, 61 male Kunming mice were randomly divided into normal group, low dose UDCA group, high dose of UDCA group, spironolactone group, and the control group.Isoproterenol (ISO) injection was given subcutaneously (30 d) to make the model of myocardial fibrosis.Corresponding anti-fibrosis drugs (UDCA or spironolactone) were given by gavage.HE staining and Masson staining were performed to explore the inflammation and fibrosis in the myocardium.The expression of collagen Ⅰ and collagen Ⅲ protein was detected by immunohistochemistry to evaluate the degree of fibrosis among the groups.Western blot was used to detect the expression of transforming growth factor, (TGF)-ß1, connective tissue growth factor (CTGF), matrix metalloproteinase (MMP)-2, -9, tissue inhibitor of metalloproteinase (TIMP)-4, -1 and anti-phospho-NFKBIA (p-IκB-α) inhibitor of NF-κB (IκB) protein in myocardium. Results: HE and Masson staining results showed that in the normal group, myocardial fibrosis is less, while the control group showed a large amount of fibrotic tissue (P<0.05). Tissue fibrosis in the low/high dose UDCA group and spironolactone group was significantly reduced compared with the control group (P<0.05), in which high dose of UDCA reduces fibrosis more significantly.Immunohistochemistry results showed that collagen Ⅰ and collagen Ⅲ protein expression was significantly increased (P<0.05). Whereas in the low/high UDCA dose group and spironolactone group, collagen Ⅰ and collagen Ⅲ expression were significantly decreased (P<0.05), the high UDCA dose group decreased more significantly.Western blot results suggest that TGFß-1 expression in the myocardial tissue was significantly increased compared to the normal group (P<0.05), whereas low/high UDCA dose group and spironolactone group, TGFß-1 protein expression were significantly decreased [UDCA(1.52±0.16), (1.02±0.12), (1.01±0.21)vs (2.73±0.12), P<0.05], in which high UDCA dose group TGFß-1 protein expression level decreased more significantly.However, there was no significant difference in the expression of CTGF, MMP2/9 and TIMP1/4 protein among the groups (P>0.05). UDCA decrease p-IκB-α expression and increase IκB protein expression dose-dependently. Conclusions: UDCA can relieve isoproterenol induced myocardial fibrosis and reduce the myocardial collagen Ⅰ and collagen Ⅲ deposition in a dose dependent manner.Down-regulating of TGFß-1 protein expression through the inhibition of TGR5-NF-κB signal transduction pathway might be a potential mechanism underlying UDCA's effects.

Lack of association between ERCC5 gene polymorphisms and gastric cancer risk in a Chinese population.

We conducted a case-control study to assess the association between single nucleotide polymorphisms in the ERCC5 promoter (rs2094258 and rs751402) and development of gastric cancer in a Chinese population. This investigation included 184 patients with pathologically diagnosed gastric cancer and 206 healthy subjects recruited between October 2012 and December 2014. The genotyping of ERCC5 rs2094258 and rs751402 variants was performed by polymerase chain reaction coupled with restriction fragment length polymorphism. Genotype distributions of these polymorphisms conformed to Hardy-Weinberg equilibrium in both patient (P = 0.25 for rs2094258 and P = 0.61 for rs751402) and control groups (P = 0.48 for rs2094258 and P = 0.42 for rs751402). Using unconditional logistic regression analysis, we found that neither of these ERCC5 variants was associated with increased risk of gastric cancer under co-dominant, dominant, or recessive models (P < 0.05). In conclusion, we suggest that the rs2094258 and rs751402 polymorphisms are not connected to the development of this disease under codominant, dominant, and recessive models.

Dietary L-arginine levels affect the liver protein turnover and alter the expression of genes related to protein synthesis and proteolysis of laying hens.

Amino acids are considered to be anabolic factors that affect protein turnover. The aim of this study was to test the effects of dietary L-arginine (Arg) levels on protein metabolism in the liver of laying hens and the expression of genes related to protein synthesis and proteolysis. Xinyang black commercial laying hens (n = 864, 31 wk of age) were randomly allotted to 6 treatments with 4 replicates of 36 birds. The dietary treatments were corn-corn gluten meal based diets containing 0.64, 0.86, 1.03, 1.27, 1.42, and 1.66% L-Arg, respectively. Serum concentrations of total protein and albumin were maximized in the 1.27% L-Arg group, and serum concentration of urea acid was the lowest in the 1.27% L-Arg group. The 1.27% L-Arg group had the highest fractional protein synthesis rate and fractional protein gain rate in the liver. Consistent with the data on protein turnover, mRNA abundances of target of rapamycin (TOR) and ribosomal protein S6 kinase 1 increased in the liver of layers fed 1.27% L-Arg, while mRNA abundances of cathepsin B and 20S proteasome decreased at the same dietary L-Arg level. In conclusion, the dietary level of L-Arg increased the liver fractional protein synthesis rate and fractional protein gain rate of laying hens, and the action of an appropriate level of dietary L-Arg involves upregulating the gene expression of the TOR signaling pathway accompanied by suppressing the mRNA expression of cathepsin B and 20S proteasome in the liver.

L-arginine upregulates the gene expression of target of rapamycin signaling pathway and stimulates protein synthesis in chicken intestinal epithelial cells.

L-arginine (Arg) is an indispensable amino acid in avians and is required for growth. The aim of this study was to investigate the effects of L-Arg on protein synthesis and genes expression involved in target of rapamycin (TOR) signaling pathway in chicken enterocytes. Cells were cultured for 4 days in L-Arg-free Dulbecco's modified Eagle's medium containing 10, 100, 200, 400, or 600 µM L-Arg. Cell growth, cell cycle, protein synthesis, and protein degradation as well as mRNA expression levels of TOR, ribosomal protein S6 kinase 1 (S6K1), and eukaryotic initiation factor 4E-binding protein 1 (4E-BP1) were determined. The results showed that cell viability was enhanced by L-Arg with a maximal response at 10 to 400 µM. Increasing extracellular concentrations of L-Arg from 10 to 400 µM increased the cells in S and G2/M phase to a significant extent and decreased cell numbers in G0/G1 phase. Further more, addition of 100, 200, or 400 µM L-Arg to culture medium increased protein synthesis and reduced protein degradation in chicken intestinal epithelial cells. Consistent with the data on cell growth and protein turnover, supplementation of 100, 200, or 400 µM L-Arg increased the mRNA abundances of TOR, 4E-BP1, and S6K1. It was concluded the action of L-Arg involves in upregulating the genes expression of TOR cell signaling pathway which increases protein synthesis and reduces protein degradation.

Isolation and characterization of novel EST-SSRs in the showy dendrobium, Dendrobium nobile (Orchidaceae).

Expressed sequence tag (EST)-derived simple sequence repeat (SSR) markers could enrich the current resource of molecular markers. In this study, microsatellite markers were developed for Dendrobium nobile Lindl. by mining the ESTs. Twenty-eight EST-SSRs amplified 2 to 6 nucleotide repeats with a mean number of 2.82 alleles per locus. The observed and expected heterozygosities per locus ranged from 0.158 to 0.579 and 0.422 to 0.752, respectively. These novel EST-SSRs enriched the current resource of molecular markers for the Dendrobium genus and would facilitate further applications in germplasm appraisal, evolution and genetic diversity studies, genetic mapping, and plant breeding of D. nobile and other congeneric species.

Enhanced Intrinsic Anomalous Valley Hall Effect Induced by Spin-Orbit Coupling in MXene Monolayer M3N2O2 (M = Y, La).

The limitation of suitable anomalous valley Hall effect (AVHE) materials has seriously hindered the booming development and the widespread application of valleytronics. Here, through the first-principles calculations, we propose a MXene monolayer Y3N2O2 with spontaneous valley polarization (VP) of 21.3 meV, which induces intrinsic AVHE. The VP can be modulated linearly, which provides a route of effective control of the valley signals. Importantly, VP can be enhanced by adjusting up the spin-orbit coupling (SOC) based on a SOC Hamiltonian model and the first-principles calculations. From this physics underlying, we substitute the Y atom with the La atom and further propose the monolayer La3N2O2, in which the heavy atom La will provide stronger SOC than Y atom. The spontaneous VP in La3N2O2 is enhanced to 100.4 meV, so AVHE can be easily achieved. Our work not only provides compelling candidates for AVHE materials but also offers a novel mindset for finding suitable valleytronic devices.

Clonal spreading of methicillin-resistant SCCmec Staphylococcus aureus with specific spa and dru types in central Taiwan.

The goal of this study was to delineate the molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) in Taiwan. Ninety-six MRSA isolates were collected from the blood cultures of different patients during the period July to December of 2008. The spa typing, staphylococcal chromosomal cassette (SCCmec) typing, mec-associated direct repeat unit (dru) copy numbers, and toxin genes (sea, seb, sec, tst, lukS/F) of each isolate were determined. Thirty-eight, 28, 18, and 12 MRSA isolates were SCCmec type II, SCCmec type III, SCCmec type IV, and SCCmec type V, respectively. Most (31/38, 81.6%) of the SCCmec type II isolates were of spa t002 with four dru repeats. Some of them also carried the sec or tst toxin gene (67.7 and 80.6%, respectively). Of the 28 SCCmec type III MRSA isolates, 15 (53.6%) were of t037 with 14 dru repeats, and all also carried the sea gene. Of the 18 SCCmec type IV MRSA isolates, 13 (72.2%) were of t437 with nine dru repeats, and ten of them also had the seb gene. Among the SCCmec type V MRSA isolates, nine were type V(T). Five (55.6%) of them were of t437 with 11 dru repeats, and all contained the lukS/F gene. The clonal spreading of SCCmec MRSA strains with specific spa and dru types was found. Further longitudinal, multiple-site surveillance is required in order to define the MRSA evolution in Taiwan.

Expression of c-FLIP in malignant melanoma, and its relationship with the clinicopathological features of the disease.

BACKGROUND: Numerous molecular events have been associated with the development of malignant melanoma (MM). The cellular FLICE-like inhibitory protein (c-FLIP) was originally identified as an inhibitor of death-receptor signalling through competition with FADD-like interleukin-1ß-converting enzyme (FLICE; also known as caspase-8) for recruitment to the FAS-associated death domain (FADD), and it has been suggested recently that c-FLIP is required for the survival and proliferation of various cell types. Aim. To investigate the relationship of c-FLIP expression with the clinicopathological features of MM. METHODS: Immunohistochemical staining with anti-c-FLIP antibody was performed on tissue samples taken from 77 MMs and 20 naevi. The proliferative cells were visualized by staining with Ki-67 antibody. Annexin V-propidium iodide labelling, a marker for chromatin condensation, was performed to observe the rate of cell apoptosis by flow cytometry. RESULTS: Expression of c-FLIP was increased in MM tissue compared with the matched pigmented naevi. The c-FLIP expression was significantly associated with the histological type and Clark level of MM, and correlated strongly with the Ki-67 labelling index. Downregulation of c-FLIP might increase apoptosis in MM cell lines. CONCLUSIONS: c-FLIP might play an important role in the obtaining of aggressive biological behaviours and be useful in predicting prognosis of patients with MM.

Genetic linkage map of EST-SSR and SRAP markers in the endangered Chinese endemic herb Dendrobium (Orchidaceae).

Dendrobium officinale is an endangered orchid from southeast Asia that is known for its medicinal properties in traditional Chinese medicine. We constructed an integrated genetic linkage map of an F(1) population derived from an interspecific cross between D. officinale and D. aduncum (both, 2n = 38), using expressed sequence tag-simple sequence repeats (EST-SSR) and sequence-related amplified polymorphism (SRAP). A total of 349 polymorphic loci, including 261 SRAP loci and 88 EST-SSR loci, were identified for genetic linkage analysis. The software JoinMap 4.0 was used to construct the genetic maps. A total of 157 loci were arranged into 27 major linkage groups, each containing a minimum of four markers, and a further 23 markers were distributed to five triplets and four doublets, the frame map covered a total distance of 1580.4 cM, with a mean of 11.89 cM between adjacent markers. This primary map of the D. officinale and D. aduncum hybrid provides a basis for genetic studies and should facilitate future studies of medical traits mapping and marker-assisted selection in Dendrobium species breeding programs.

Molecular evolution of two consecutive carotenoid cleavage dioxygenase genes in strigolactone biosynthesis in plants.

Strigolactones are newly discovered plant hormones that perform various functions, from signaling in symbiotic interactions with arbuscular mycorrhizal fungi to controlling outgrowth of axillary buds. We examined the phylogenetic relationships of two carotenoid cleavage dioxygenase genes (CCD7 and CCD8) that are involved in consecutive upstream steps of the proposed strigolactone biosynthesis pathway. The CCD7 and CCD8 sequences from 11 model species, divided into two clades, correspond to sequences from monocotyledons and dicotyledons. However, the sequences from the primitive moss, Physcomitrella patens, appeared to be evolutionarily distinct from those of the angiosperms. CCD7 and CCD8 are much conserved, since no significant positive selection was detected among these plants. Ks values indicated that CCD7 and CCD8 diverged about 290 to 430 million years ago. As essential genes in the strigolactone pathway, the divergence timing of the conserved CCD7 and CCD8 genes reflects the approximate time of generation of strigolactone as a regulatory substance. This timing calculation also coincides with initiation of symbiosis between plants and microorganisms, inferred from the fossil record. Molecular evolution analyses of genes in metabolic pathways can provide insight concerning gene evolution.