High prevalence of vertebral deformity in tumor-induced osteomalacia associated with impaired bone microstructure.

PURPOSE: Patients with tumor-induced osteomalacia (TIO) often suffer from irreversible height loss due to vertebral deformity. However, the prevalence of vertebral deformity in TIO patients varies among limited studies. In addition, the distribution and type of vertebral deformity, as well as its risk factors, remain unknown. This study aimed to identify the prevalence, distribution, type and risk factors for vertebral deformity in a large cohort of TIO patients. METHODS: A total of 164 TIO patients were enrolled in this retrospective study. Deformity in vertebrae T4-L4 by lateral thoracolumbar spine radiographs was evaluated according to the semiquantitative method of Genant. Bone microstructure was evaluated by trabecular bone score (TBS) and high-resolution peripheral QCT (HR-pQCT). RESULTS: Ninety-nine (99/164, 60.4%) patients had 517 deformed vertebrae with a bimodal pattern of distribution (T7-9 and T11-L1), and biconcave deformity was the most common type (267/517, 51.6%). Compared with patients without vertebral deformity, those with vertebral deformity had a higher male/female ratio, longer disease duration, more height loss, lower serum phosphate, higher bone turnover markers, lower TBS, lower areal bone mineral density (aBMD), lower peripheral volumetric BMD (vBMD) and worse microstructure. Lower trabecular vBMD and worse trabecular microstructure in the peripheral bone and lower spine TBS were associated with an increased risk of vertebral deformity independently of aBMD. After adjusting for the number of deformed vertebrae, we found little difference in clinical indexes among the patients with different types of vertebral deformity. However, we found significant correlations of clinical indexes with the number of deformed vertebrae and the spinal deformity index. CONCLUSION: We reported a high prevalence of vertebral deformity in the largest cohort of TIO patients and described the vertebral deformity in detail for the first time. Risk factors for vertebral deformity included male sex, long disease duration, height loss, abnormal biochemical indexes and bone impairment. Clinical manifestation, biochemical indexes and bone impairment were correlated with the number of deformed vertebrae and degree of deformity, but not the type of deformity.

Efficacy and safety of endothelin receptor antagonists in type 2 diabetic kidney disease: A systematic review and meta-analysis of randomized controlled trials.

AIM: To analyse the efficacy and safety of endothelin receptor antagonists for people with diabetic kidney disease. METHODS: Randomized controlled trials comparing endothelin receptor antagonists with placebo in people with diabetic kidney disease were identified through PubMed, Embase and the Cochrane Library. We used a random-effect model to calculate the mean difference or risk ratio with the 95% CI. RESULTS: Seven studies with a total of 4730 participants were included. Overall, endothelin receptor antagonists significantly reduced albuminuria compared with placebo (standardized mean difference -0.48, 95% CI -0.64 to -0.33). Atrasentan, in particular, effectively reduced albuminuria (standardized mean difference -0.58, 95% CI -1.00 to -0.17) and the risk of composite renal endpoints (risk ratio 0.65; 95% CI 0.49 to 0.88), with insignificant change in the rate of congestive heart failure (risk ratio 1.40, 95% CI 0.76 to 2.56) and mortality (risk ratio 1.11, 95% CI 0.77 to 1.61). In contrast, although avosentan reduced albuminuria (standardized mean difference -0.47, 95% CI -0.57 to -0.36) and the risk of composite renal endpoints (risk ratio 0.63, 95% CI 0.42 to 0.94), it was associated with a significant increase in congestive heart failure risk (risk ratio 2.61, 95% CI 1.36 to 5.00) and an insignificant increase in mortality risk (risk ratio 1.50, 95% CI 0.81, 2.78). No significant change in efficacy or safety outcomes with bosentan was detected. Dose-response analysis indicated that 0.75 mg/day atrasentan is expected to be optimal for renoprotection, with maximal albuminuria reduction and minimal fluid retention events. CONCLUSIONS: Among the endothelin receptor antagonists, atrasentan and avosentan, but not bosentan, are effective for renoprotection in people with diabetic kidney disease. Compared with other types and doses, atrasentan 0.75 mg/day is the most promising, with maximal albuminuria reduction and minimal fluid retention. Vigilant monitoring of congestive heart failure risk is needed in future clinical practice. (PROSPERO registration no. CRD42020169840).

Noninvasive prenatal diagnosis for pregnancies at risk for ß-thalassaemia: a retrospective study.

OBJECTIVE: To evaluate the clinical feasibility of noninvasive prenatal diagnosis (NIPD) for ß-thalassaemia using circulating single molecule amplification and re-sequencing technology (cSMART). DESIGN: Through carrier screening, 102 pregnant Chinese couples carrying pathogenic HBB gene variants were recruited to the study. Pregnancies were managed using traditional invasive prenatal diagnosis (IPD). Retrospectively, we evaluated the archived pregnancy plasma DNA by NIPD to evaluate the performance of our cSMART assay for fetal genotyping. SETTING: Chinese prenatal diagnostic centres specialising in thalassaemia testing. POPULATION: Chinese carrier couples at high genetic risk for ß-thalassaemia. METHODS: Fetal cell sampling was performed by amniocentesis and HBB genotypes were determined by reverse dot blot. NIPD was performed by a newly designed HBB cSMART assay and fetal genotypes were called by measuring the allelic ratios in the maternal cell-free DNA. MAIN OUTCOME MEASURES: Concordance of HBB fetal genotyping between IPD and NIPD and the sensitivity and specificity of NIPD. RESULTS: Invasive prenatal diagnosis identified 29 affected homozygotes or compound heterozygotes, 54 heterozygotes and 19 normal homozygotes. Compared with IPD results, 99 of 102 fetuses (97%) were correctly genotyped by our NIPD assay. Two of three discordant samples were false positives and the other sample involved an incorrect call of a heterozygote carrier as a homozygote normal. Overall, the sensitivity and specificity of our NIPD assay was 100% (95% CI 88.06-100.00%) and 97.26% (95% CI 90.45-99.67%), respectively. CONCLUSIONS: This study demonstrates that our cSMART-based NIPD assay for ß-thalassaemia has potential clinical utility as an alternative to IPD for pregnant HBB carrier couples. TWEETABLE ABSTRACT: A new noninvasive test for pregnancies at risk for ß-thalassaemia.

PAX3 silencing suppresses gastric cancer proliferation and angiogenesis via MET/PI3K signaling.

PAX3 is the key factor in cell signal transduction pathway and may be involved in the regulation of cancer cell proliferation, differentiation and migration. The aim of the study was to investigate the effects and mechanism of PAX3 silencing on the gastric cancer. Specific PAX3 silencing was performed both in vitro and in vivo using small-interfering RNAs (siRNAs). The proliferation, apoptosis and angiogenesis of gastric cancer cells were assessed using MTT assay, flow cytometry and in vitro tube formation assay. Mice with gastric xenografts, which expressed either si-PAX3 or non-coding siRNA (si-NC), were developed and the effects of PAX3 silencing on tumor progression were evaluated. PCNA is a proliferating cell nuclear antigen and can be used as an index for evaluating cell proliferation status. Immunocytochemistry assay was used to quantify PAX3 and PCNA expression. After 4 weeks of tumor inoculation, tumor tissues were weighed. Tumor tissue morphology and apoptosis were evaluated using HE staining and TUNEL assay. In order to investigate the effect of silencing PAX3 on cell apoptosis, angiogenesis and MET/PI3K pathway, quantitative real-time PCR (qRT-PCR) or western blot were used to detect the expression levels of caspase-3, VEGF, MET, p-MET, PI3K and p-PI3K. After PAX3 silencing, PAX3 expression was significantly decreased in two gastric cancer cell lines, MKN-28 and SGC-7901 (p<0.05 vs Control). PAX3 silencing reduced cell proliferation, induced cell apoptosis and inhibited tube formation. PAX3 and PCNA expression were also significantly decreased. In mice, silencing PAX3 significantly inhibited tumor growth and decreased microvessel density in tumor. PAX3 silencing also decreased cell density in tumors, which concurred with increased apoptosis and PAX3 expression. PAX3 silencing upregulated the expression of caspase-3, downregulated the expression of VEGF, phosphorylation of PI3K and MET. Our data showed that these anti-tumor effects of PAX3 silencing might be attributed to its role in inducing cell apoptosis and inhibiting angiogenesis.

Nasal fluid cytology and cytokine profiles of eosinophilic and non-eosinophilic chronic rhinosinusitis with nasal polyps.

BACKGROUND: Chronic rhinosinusitis with nasal polyps (CRSwNP) is a heterogeneous disease with different clinical characteristics and different treatment responsiveness. The aims of this study were to compare the nasal fluid cytology and cytokines between eosinophilic CRSwNP (eCRSwNP) and non-eosinophilic CRSwNP (neCRSwNP) and establish a new multivariate model to predict eCRSwNP before surgery to improve personalized treatment for CRSwNP patients. METHODS: Eighty-six consecutive patients with CRSwNP and sixteen healthy controls were recruited in this study. Nasal fluid (NF) was collected from all subjects and nasal polyp tissue was collected during the surgery. The differential cell counts and concentrations of IL-6, IL-8, TNF-77; and IL-10 in NF were measured. Univariate and multivariate logistic regression were used to identify predictors for eCRSwNP. RESULTS: There were more inflammatory cells in NF of CRSwNP than controls. The eosinophil percentage was significantly higher in eCRSwNP than neCRSwNP and controls. The level of IL-8 was significantly higher in neCRSwNP than in eCRSwNP and controls. Blood eosinophilia, nasal fluid eosinophilia, higher total ethmoid score / total maxillary score (E/M ratio) and higher visual analogue scale (VAS) score of CRS were associated with eCRSwNP, the area under receiver operating characteristic curve (AUC) was 0.800, 0.755, 0.703 and 0.648, respectively. Using the coefficients of multivariate regression, we set up a scoring system to predict eCRSwNP with three of the variates and the AUC was 0.883. CONCLUSION: ECRSwNP, neCRSwNP and healthy controls demonstrated different cytology and cytokine profiles in NF. A new preoperational multivariate prediction model for eCRSwNP with NF eosinophilia, blood eosinophilia and higher E/M ratio was established.

Noninvasive fetal genotyping in pregnancies at risk for PKU using a comprehensive quantitative cSMART assay for PAH gene mutations: a clinical feasibility study.

OBJECTIVE: To assess the diagnostic performance of a novel circulating single molecule amplification and re-sequencing technology (cSMART) method for noninvasive prenatal testing (NIPT) of Phenylketonuria (PKU). DESIGN: Blinded NIPT analysis of pregnancies at high risk for PKU. SETTING: Shanghai Xinhua Hospital and Hunan Jiahui Genetics Hospital, China. POPULATION: Couples (n = 33) with a child diagnosed with PKU. METHODS: Trio testing for pathogenic PAH mutations was performed by Sanger sequencing. In second pregnancies, invasive prenatal diagnosis (IPD) was used to determine fetal genotypes. NIPT was performed using a PAH gene-specific cSMART assay. Based on the plasma DNA mutation ratio relative to the fetal DNA fraction, fetal genotypes were assigned using a maximum-likelihood algorithm. MAIN OUTCOME MEASURES: Concordance of fetal genotyping results between IPD and NIPT, and the sensitivity and specificity of the NIPT assay. RESULTS: Compared with gold standard IPD results, 32 of 33 fetuses (96.97%) were accurately genotyped by NIPT. The sensitivity and specificity of the NIPT assay was 100.00% (95% CI 59.04-100.00%) and 96.15% (95% CI 80.36-99.90%), respectively. CONCLUSIONS: The novel cSMART assay demonstrated high accuracy for correctly calling fetal genotypes. We propose that this test has useful clinical utility for the rapid screening of high-risk and low-risk pregnancies with a known history of PKU on one or both sides of the family. TWEETABLE ABSTRACT: NIPT of couples at high risk for PKU using a full-coverage cSMART PAH gene test.

[Effects of sulfur dioxide on alveolar macrophage apoptosis in acute lung injury induced by limb ischemia/reperfusion in rats].

OBJECTIVE: To investigate the effect of sulfur dioxide (SO2) on the apoptosis of alveolar macrophage (AM) in lung protection of limb ischemia/reperfusion (I/R) induced acute lung injury (ALI), and to find a new target for the control of inflammatory response. METHODS: Twenty pathogen-free, adult male Sprague-Dawley (SD) rats (180-230 g) were used in this study. Five rats were to be used for limb ischemia/reperfusion, then plasma was extracted as ischemia/reperfusion serum stimulation. Fifteen rats were to be used for extracting AM by bronchoalveolar lavage. The AM was isolated and cultured, then the cell count was adjusted to 1×106/mL, and randomly divided into the following 4 groups (n=6): control group, I/R group, SO2 group, and I/R+SO2 group. The I/R group was given ischemia/reperfusion serum (500 µg/L) to stimulate 6 h; the SO2 group was given an SO2 donor, Na2SO3/NaHSO3 [(0.54 mmol/kg) / (0.18 mmol/kg)]; and the I/R+SO2 group was given the same ischemia/reperfusion serum and Na2SO3/NaHSO3 at the same time. The level of mitochondrial membrane potential, the state of mitochondrial permeability transition pore (mPTP), the rate of AM apoptosis, the expression of Bcl-2 and Caspase-3 proteins were detected by flow cytometry, microplate reader and Western blotting. RESULTS: Compared with the control group, in the I/R group, the ratio of red to green fluorescence and the absorbance decreased significantly, the percentage of apoptotic cells increased obviously, the apoptotic rate was 43.81%±2.40%, Caspase-3 protein expression increased, Bcl-2 protein expression decreased. While compared with the I/R group, in the I/R+SO2 group, the ratio of red to green fluorescence and the absorbance increased significantly; the apoptotic rate decreased to 37.01%±1.93%, Caspase-3 protein expression decreased, Bcl-2 protein expression increased. CONCLUSION: Exogenous SO2 has the effect of accelerating AM apoptosis by stimulating mPTP to open and mitochondrial membrane potential to decrease; besides, exogenous SO2 could stimulate AM to secrete more anti-inflammatory cytokines and less inflammatory cytokines. In conclusion, exogenous SO2 can reduce macrophage apoptosis by inhibiting mitochondrial pathways.

Long-term follow-up and treatment of a patient with severe skeletal open bite using temporary anchorage devices.

This case report presents the treatment and long-term follow-up of a patient with severe skeletal hyperdivergent open bite, Class II malocclusion, and a severely retruded chin. After failure of early treatment using high-pull headgear with a bite block during the early permanent dentition stage due to an unfavorable growth pattern, orthognathic surgery was proposed but rejected by the patient. Then, temporary anchorage devices were used to correct the occlusion and establish an acceptable overbite and overjet. The overall observation time was 8.5 years; the treatment time using fixed appliances was 3 years and 4 months. The achieved tooth position and occlusal relationship remained stable 2.5 years later without recurrence of the open bite.

Sclerosing stromal tumor of the ovary in a perimenopausal woman: a case report.

Sclerosing stromal tumor (SST) is a rare ovarian neoplasia deriving from the sex cord stromal tumor, which occurs usually in the sec- ond and third decades of life. However, the authors report a 46-year old multiparous woman who presented with a cystic-solid left pelvic mass, a large amount of ascites, and elevated serum CA-125, all suggesting a malignant tumor. Surgery was performed and final histopathological diagnosis of the specimen was diagnosed with SST. The authors herein report an extremely rare case of SST with a cystic-solid pelvic mass and a large amount of ascites, which is useful to demonstrate the possibility of SST in multiparous woman.

TNF-α was involved in calcium hydroxide-promoted osteogenic differentiation of human DPSCs through NF-κB/p38MAPK/Wnt pathway.

It has been reported that calcium hydroxide can induce proliferation, migration, and mineralization in dental pulp stem cells (DPSCs), but the underlying molecular mechanisms are still unclear. In this study, we sought to explore the role of calcium hydroxide in the cell proliferation and directional differentiation of DPSCs and to study the regulatory effect of NF-κB, p38MAPK, and Wnt signaling on differentiation of DPSCs. CCK8 cell assay, Wound Healing Assay, and Alkaline Phosphatase Staining Assay were respectively used to determine the proliferation rate, migration and ALP expression of DPSCs. Alizarin Red Staining Assay was used to observe the mineralization of DPSCs. RT-PCR analysis and Western Blot Analysis displayed the expression of related fators at mRNA and protein level, respectively. In the present study, we found that NF-κB, p38MAPK, and Wnt signaling could abolish calcium hydroxide-induced proliferation of DPSCs. The inhibition of NF-κB, p38MAPK, and Wnt signaling suppressed the migration, ALP expression, and mineralization of DPSCs. NF-κB, p38MAPK, and Wnt signaling involved in directional differentiation of DPSCs. Moverover, calcium hydroxide could activate NF-κB, p38MAPK, and Wnt pathway by regulating TNF-α. Our study showed that NF-κB, p38MAPK, and Wnt signaling pathway were involved in calcium hydroxide-induced proliferation, migration, mineralization, and osteogenic differentiation in DPSCs. Calcium hydroxide affected NF-κB, p38MAPK, and Wnt pathway by regulating TNF-α.

Placental site trophoblastic tumor: A review of 108 cases and their implications for prognosis and treatment.

OBJECTIVE: To identify important prognostic factors and optimized treatment strategies through the analysis of the clinical and pathological characteristics of placental site trophoblastic tumor. METHODS: 108 patients with PSTT registered in two GTD centers or in six tertiary hospitals in China were analyzed retrospectively between the years 1998 and 2013. The computerized database of clinical and pathological reports was reviewed on this patient group. The data were subsequently analyzed retrospectively using SPSS software. RESULTS: Among 3581 patients with GTNs treated in GTD centers or in the tertiary hospitals between 1998 and 2013, 108 cases were histologically confirmed PSTT (3%). Only seven deaths and eleven relapse cases were observed. All seven of the deaths were disease related, due to chemotherapy-resistant or relapsed. 23 patients who received fertility preservation treatment did not experience poor outcome or high risk of relapse. In 71 patients with International Federation of Gynecology and Obstetrics (FIGO) stage I disease, the use of adjuvant chemotherapy following surgery (n=49) or not (n=22) made no significant difference in relapse rate (P=0.303) or survival (P=0.782). Univariate analysis revealed the interval between antecedent pregnancy and onset of PSTT, stage, prognosis score, and necrosis as significant predictors of poor survival but only stage remained significant on multivariate analysis. CONCLUSIONS: Patients with FIGO stage IV disease demonstrate the most critical risk indicator of PSTT in the current study. Preservation of fertility is considered in highly-selected patients with localized tumor; and surgery without chemotherapy is recommended as first line treatment for patients with stage I who are at low-risk.

A laparoscopic surgery for deep infiltrating endometriosis and the review of literature.

Deep infiltrating endometriosis (DIE) is a complex disorder that affects 6% to 12% of all women in the reproductive age. In these cases, treatment is more difficult with possible incomplete pain relief and a considerable possibility of recurrence. Here, the authors report a case of a 41-year-old woman with a history of severe dysmenorrhea, dyspareunia, and chronic pelvic pain because of deep infiltrating pelvic and peritoneal endometriosis, who underwent segmental colorectal resection three years ago for large bowel obstruction due to colonic endometriosis. To ensure complete removal of the disease, the authors injected gonadotropin-releasing hormone agonist (GnRHa) in three periodic cycles before laparoscopic surgery. We performed laparoscopic hysterectomy and deep pelvic nodule resection and pelvic adhesion releasing. After five days of hospitalization, the patient recovered totally and was not experiencing any pain at three months' follow-up. Laparoscopic treatment has more become the standard of treatment for DIE. A review of the literature regarding pathology and physiology of DIE and surgical aspects of its management is undertaken. The authors would like to renew the current laparoscopic surgery in curing the DIE, as they believe that this is also a useful addition to the literature.

Epidemiology of chronic rhinosinusitis: results from a cross-sectional survey in seven Chinese cities.

BACKGROUND: Chronic sinusitis (CRS) is a common otorhinolaryngologic disease that is frequently encountered in everyday practice, but there is a lack of precise data regarding the prevalence of CRS in developing countries. We performed a national investigation in China to determine the prevalence and associated factors of CRS. METHODS: We conducted a cross-sectional investigation in 2012. A stratified four-stage sampling method was used to select participants randomly from seven cities in mainland China. All participants were interviewed face-to-face via a standardized questionnaire. Unconditional logistic regression analyses were conducted to examine the association between smoking and sinusitis after adjusting for socio-demographic factors. RESULTS: This study included a total of 10 636 respondents from seven cities. The overall prevalence of CRS was 8.0% and ranged from 4.8% to 9.7% in seven centres. Chronic sinusitis affected approximately 107 million people in mainland China. Chronic sinusitis was particularly prevalent among people with specific medical conditions, including allergic rhinitis, asthma, chronic obstructive pulmonary disease and gout. The prevalence was slightly higher among males (8.79%) than females (7.28%) (P = 0.004), and the prevalence varied by age group, ethnicity and marital status and education (P < 0.05), but not by household per capita income or living space (P > 0.05). Both second-hand tobacco smoke and active smoking were independent risk factors for CRS (P = 0.001). CONCLUSIONS: Chronic sinusitis is an important public health problem in China. Our study provides important information for the assessment of the economic burden of CRS and the development and promotion of public health policies associated with CRS particularly in developing countries.

Serum 25-hydroxyvitamin D levels and peripheral neuropathy in patients with type 2 diabetes: a systematic review and meta-analysis.

BACKGROUND: Vitamin D is a multifunctional pro-hormone and has widespread actions in human body. Several studies showed a possible association between vitamin D deficiency and diabetic peripheral neuropathy (DPN) in patients with type 2 diabetes, but no definite conclusion was available. METHODS: A systematic review and meta-analysis was performed to comprehensively assess the association between serum 25-hydroxyvitamin D [25(OH)D] levels and DPN in patients with type 2 diabetes. Data from eligible studies were pooled using meta-analysis. RESULTS: Six studies that involved a total of 1,484 type 2 diabetic patients were finally included into the meta-analysis. Meta-analysis showed that there were obviously decreased serum 25(OH)D levels in DPN patients [weighted mean difference (WMD) = -6.36 ng/ml, 95 % confidence interval (95 % CI) -8.57 to -4.14, P < 0.00001]. Vitamin D deficiency was also significantly associated with increased risk of DPN in patients with type 2 diabetes [odds ratio (OR) 2.88, 95 % CI 1.84-4.50, P < 0.00001]. Meta-analysis of three studies with adjusted estimates showed that vitamin D deficiency was independently associated with increased risk of DPN in patients with type 2 diabetes (OR 2.68, 95 % CI 1.67-4.30, P < 0.0001). Sensitivity analysis showed that there was no obvious change in the pooled estimates. CONCLUSION: Vitamin D is involved in the development of DPN in type 2 diabetic patients, and vitamin D deficiency is very likely to be associated with DPN in type 2 diabetic patients. Further studies are needed to validate the association between vitamin D deficiency and DPN.

Analysis of the expression of HMGB-1, CXCL16, miRNA-30a, and TGF-ß1 in primary nephritic syndrome patients and its significance.

We investigated the expression levels of high-mobility group box protein 1 (HMGB-1), CXC chemokine ligand 16 (CXCL16), microRNA (miRNA)-30a and transforming growth factor ß1 (TGF-ß1) in primary nephritic syndrome (PNS) patients and the clinical significance of this expression. A total of 56 patients with PNS were included in the PNS group, while 50 healthy subjects formed the normal control group. Serum levels of HMGB-1, CXCL16, miRNA-30a, and urinary TGF-ß1 concentrations were quantified along with other biochemical indices, including serum albumin, triglyceride (TG), total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein, and urinary proteins. The correlation between levels of HMGB-1, CXCL16, miRNA-30a, and TGF-ß1 and biochemical indexes was further analyzed. PNS group patients had significantly higher levels of HMGB-1, CXCL16, miRNA- 30a, and TGF-ß1 compared to the control group (P < 0.05). PNS patients also had higher 24-h urinary protein, TG, TC, and LDL levels but lower serum albumin compared to subjects in the control group (P < 0.05). Serum HMGB-1, CXCL16, miRNA-30a, and urinary TGF-ß1 levels were all negatively correlated with serum albumin levels, but were positively correlated with TG, TC, LDL, and 24-h urinary protein (P < 0.05 in all cases). Additionally, a positive correlation existed among serum HMGB-1, CXCL16, miRNA-30a, and urinary TGF-ß1 levels (P < 0.01). HMGB-1, CXCL16, miRNA-30a, and urinary TGF-ß1 were highly expressed in PNS patients and may play important roles in the pathogenesis and development of PNS.

Elucidating the function and tolerance mechanism of gamma delta (γ δ) T cells in a Helicobacter pylori infection model.

In this study, the functions and mechanisms of γ δ T cells were analyzed in patients infected with Helicobacter pylori. Peripheral blood was collected from gastritis patients in the Gastroenterology Department of Ningbo No. 2 Hospital. Preliminary analyses revealed 24 H. pylori-positive and 17 H. pylori-negative patients. The wild-type and γ δ T knockout mice were infected with cultured H. pylori cells (obtained from the H. pylori-positive patients). H. pylori in mice was quantified by polymerase chain reaction; gastritis was confirmed by hematoxylin and eosin staining. The TCR-δ(-/-) mice were treated with vein adoptive immunotherapy 24 h prior to H. pylori inoculation; the same method was used to detect the extent of gastritis and bacterial colonization. The γ δ T knockout mice showed high levels of H. pylori infection than the wild-type mice; in addition, the knockout mice showed severe disease pathology. γ δ T knockout mice also displayed increased matrix metalloproteinase-9 (MMP-9) and decreased MMP-7 expression in the gastric mucosa. γ δ T cells play a protective role in patients infected with H. pylori. γ δ T cell [responsible for the production of interleukin-17 (IL-17) and IL-22] expression was increased in H. pylori-positive patients, indicating statistical significance. However, there was no significant difference in interferon-gamma + γ δ T expression between the positive and negative patients. This study demonstrated the probable involvement of γ δ T cells in the immune response of an organism, via the secretion of IL-17 and IL-22.

Characteristics of Cyclin B and its potential role in regulating oogenesis in the red claw crayfish (Cherax quadricarinatus).

Cyclin B is a regulatory subunit of maturation-promoting factor (MPF), which has a key role in the induction of meiotic maturation of oocytes. MPF has been studied in a wide variety of animal species; however, its expression in crustaceans is poorly characterized. In this study, the complete cDNA sequence of Cyclin B was cloned from the red claw crayfish, Cherax quadricarinatus, and its spatiotemporal expression profiles were analyzed. Cyclin B cDNA (1779 bp) encoded a 401 amino acid protein with a calculated molecular weight of 45.1 kDa. Quantitative real-time PCR demonstrated that Cyclin B mRNA was expressed mainly in the ovarian tissue and that the expression decreased as the ovaries developed. Immunofluorescence analysis revealed that the Cyclin B protein relocated from the cytoplasm to the nucleus during oogenesis. These findings suggest that Cyclin B plays an important role in gametogenesis and gonad development in C. quadricarinatus.

Molecular cloning, characterization and expression analysis of a Doublesex gene from Daphnia carinata (Crustacea: Cladocera) during different reproductive stages.

To better understand the reproductive transformation mechanism of Daphnia carinata, a Doublesex (Dsx) gene was cloned based on rapid amplification of cDNA ends (RACE), and was designated DapcaDsx2. Next, we compared similarities and assumed homology based on deduced amino acid sequences. It showed 97.52, 87.94, and 85.11% identity to orthologous genes in D. magna, D. pulex, and D. galeata respectively. Phylogenetic analysis revealed that DapcaDsx2 clustered in the same class, and was evolutionarily more distant to sequences from other species. qRT-PCR showed that DapcaDsx2 was most abundantly expressed during sexual reproduction (P < 0.05). Using digoxigenin-labeled RNA probes, we studied DapcaDsx2 expression in parthenogenetic and sexual females by whole-mount in situ hybridization. The results revealed that DapcaDsx2 was mainly expressed in the second antennae and several sites of the ventral carapace, whereas higher expression levels were found in sexual than in parthenogenetic females. This suggests that the DapcaDsx2 gene is involved in switching modes of reproduction and in sexual differentiation.

Ipriflavone reverses the adverse effects of a low-calcium diet on the histology of the tibia in caged layers.

1. Ipriflavone (IP) has been widely studied in humans and is effective for inhibiting osteoclastic bone resorption and enhancing osteoblastic bone formation. The aim of this study was to investigate the effects of IP on the structural histology and histomorphometric parameters of tibia in laying hens fed on a low-calcium diet. 2. A total of 200 Hy-Line Brown laying hens, 24 weeks of age, were divided randomly into 4 groups which were fed on one of 4 diets (CaN, standard diet; CaL, low-calcium diet; IP1, low-calcium diet + 8 mg/kg of IP; IP2, low-calcium diet + 20 mg/kg of IP) for 60 d after which 10 hens of each group, chosen at random, were killed and the tibiae were processed and assessed under a fluorescent microscope 3. The cortical bone showing resorption cavities had become porous, and medullary bone did not completely fill the marrow cavity in the CaL group compared with the CaN group. However, in the IP2 group, the cortical bone showed a structure comparable to the CaN group with an absence of resorption cavities, and more medullary bone filled the marrow cavity compared with the CaL group. 4. The CaL hens had significantly lower trabecular bone area, trabecular perimeter, per cent trabecular area, trabecular number and trabecular thickness and significantly wider trabecular separation compared with CaN. These results indicated that the low-calcium diet led to increased loss of tibial bone, eventually causing osteoporosis. However, compared with the CaL group, the IP2 group caused a significant increase in trabecular bone area and bone quality. 5. It is concluded that even in hens fed on a low-calcium diet, the addition of 20 mg/kg IP exerted beneficial effects on histological structure and structural histomorphometric parameters of tibia, indicating an alleviation of caged layer osteoporosis (CLO).

Intestinal Translocation of Live Porphyromonas gingivalis Drives Insulin Resistance.

Periodontitis has been emphasized as a risk factor of insulin resistance-related systemic diseases. Accumulating evidence has suggested a possible "oral-gut axis" linking oral infection and extraoral diseases, but it remains unclear whether periodontal pathogens can survive the barriers of the digestive tract and how they play their pathogenic roles. The present study established a periodontitis mouse model through oral ligature plus Porphyromonas gingivalis inoculation and demonstrated that periodontitis aggravated diet-induced obesity and insulin resistance, while also causing P. gingivalis enrichment in the intestine. Metabolic labeling strategy validated that P. gingivalis could translocate to the gastrointestinal tract in a viable state. Oral administration of living P. gingivalis elicited insulin resistance, while administration of pasteurized P. gingivalis had no such effect. Combination analysis of metagenome sequencing and nontargeted metabolomics suggested that the tryptophan metabolism pathway, specifically indole and its derivatives, was involved in the pathogenesis of insulin resistance caused by oral administration of living P. gingivalis. Moreover, liquid chromatography-high-resolution mass spectrometry analysis confirmed that the aryl hydrocarbon receptor (AhR) ligands, mainly indole acetic acid, tryptamine, and indole-3-aldehyde, were reduced in diet-induced obese mice with periodontitis, leading to inactivation of AhR signaling. Supplementation with Ficz (6-formylindolo (3,2-b) carbazole), an AhR agonist, alleviated periodontitis-associated insulin resistance, in which the restoration of gut barrier function might play an important role. Collectively, these findings reveal that the oral-gut translocation of viable P. gingivalis works as a fuel linking periodontitis and insulin resistance, in which reduction of AhR ligands and inactivation of AhR signaling are involved. This study provides novel insight into the role of the oral-gut axis in the pathogenesis of periodontitis-associated comorbidities.