Objective To clarify the role of claudin-4 in endometrial tumorigenesis and explore claudin-4 be as potentially useful agent in the treatment of endometrial carcinoma.Methods The expression of claudin-4 in 62 endonetrioid endometrial carcinoma (EEC),30 atypical hyperplasia endometrial tissue and 60 human normal endometrium was determined using immunohistochemistry and realtime PCR.Ninety female BALB/c mice were transplanted with Ishikawa endometrial cancer cells,which were divided into three groups with different intraperitoneal treatments with cisplatin,paclitaxel and saline solution.After the observation period,the tumors were extracted and stained with monoclonal antibody against claudin-4.The messenger RNA expression of claudin-4 was also detected using real-time PCR.Results Among the EEC samples,34％ (21/62) showed medium staining for claudin-4 and 66％ (41/62) showed intense staining.In atypical hyperplasia group,27％ (8/30) showed weak staining,53％ (16/30) showed medium staining and 20％ (6/30) showed intense staining for claudin-4.Of the normal endometrial tissue,47％ (28/60) showed weak staining and 53％ (32/60) showed no staining for claudin-4.According to real-time PCR,the relative quantity of claudin-4 was 170 ± 12 in EEC group,89 ± 15 in atypical hyperplasia group and 18 ± 3 in normal endometrium.Compared with those in atypical hyperplasia group and normal endometrium group,the protein and mRNA expression of claudin-4 were significantly increased in the group of EEC (all P ＜ 0.05).In the study of Ishikawa xenografts,no significant changes in tumor volume and claudin-4 expression were shown in paclitaxel group compared with that in the control group.Nevertheless,a significant reduction of the tumor growth and a significant decrease in claudin-4 expression were observed in cisplatin group.After cisplatin treatment,the tumor volume was significantly decreased [(0.51 ±0.21) versus (0.73 ±0.12) cm3],and the mRNA expression of claudin-4 was also significantly decreased (153 ± 35 versus 273 ± 27).Conclusion These results demonstrate that claudin-4 is strongly expressed in EEC,which may be a useful biomarker to monitor the effects of chemotherapy in patients with endometrial carcinoma.

Objective To detect the expression of GHR in colorectal cancer cell lines and determine whether recombinant human growth hormone can promote the proliferation of colorectal cancer cells in vitro.Methods GHR distribution was assessed by flow cytometry and immunofluorescence method in 9 colorectal cancer cell lines.The effect of recombinant human growth factor on colorectal cancer cell line proliferation was assessed by MTT method.Results Different GHR expression was determinated in 9 colorectal caner cell lines.GHR was highly expressed in HCT-8 while GHR expression could hardly be detected in LoVo.r-hGH could promote GHR(+) HCT-8 cell proliferation at 50 ng/ml and 100 ng/ml (P ＜0.05).But this effect was not dose dependent.When the neutralizing antibody was used to block GHR activity,this r-hGH dependent proliferation effect was eliminated.r-hGH could not promote GHR (-) LoVo cell proliferation (P ＞0.05).Conclusion The results demonstrates that r-hGH could promote GHR (+) tumor cell proliferation and this effect is mediated by GHR.The use of r-hGH on the colorectal cancer patients should be cautious.

Objective:To investigate the effect of verapamil on the endotoxin-induced cytokine production and nuclear factor kappa B(NF-?B) activation in the liver. Methods:Fifty six adult male Sprague-Dawley rats were randomly divided into seven groups: group A: saline;group B:endotoxin(10 mg/kg,intraperitoneally,i.p.);group C,D,E,F:endotoxin(10 mg/kg) after treatment with verapamil(1,2.5,5,10 mg/kg i.p.respectively),and group G:verapamil alone(10 mg/kg).Tumor necrosis factor(TNF-?),interleukin-6(IL-6), interleukin-10(IL-10) and NF-?B in the liver tissues and the serum levels of ALT and AST were investigated at 1 h afer LPS injection. Results:Endotoxin stimulated production of TNF-?,IL-6 and IL-10,and activated NF?B in the liver.Verapamil attenuated acute liver injury,down-regulated endotoxin-induced pro-inflammatory cytokine production,up-regulated ant-inflammatory cytokines production and inhibited NF-?B activation in the liver in a dose-dependent manner.Conclusion:Verapamil can attenuate acute liver injury by down-regulating the production of TNF-?,IL-6 and up-regulating IL-10 in the liver in a dose-dependent manner,possibly via inhibition of NF-?B.

Objective To test the effect of recombinant human growth hormone (rhGH) on the radiotherapy sensitivity of colorectal cancer cell line, and to explore its relationship with apoptosis. Methods Flow cytometry and immunofluorescence were used to detect growth hormone receptor(GHR) expression on 9 human colorectal cancer cell lines. The colony forming assay was performed to measure the post-radiotherapy colorectal cancer cell proliferation as an indicator of radiotherapy sensitivity. Flow cytometry (Annexin V-FITC staining) was used to detect the radiotherapy induced apoptosis; Westem blot was performed to detect the phosphorylated Akt level within the same cell lines. Results HCT-8 GHR positive expression cell and LoVo GHR negative expression cells were selected for further studies. The colony formation rate was significantly enhanced in HCT-8 cells pre-incubated with thGH as compared to the radiotherapy group ceUs and in a dose dependent manne(0-100 mg/L); under high doses (8 Gy), this effect was more dramatic (52.1±2.9 vs 21.0±2.7, P<0.001). thGH pre-incubation also reduced radiotherapy induced HCT-8 cell apoptosis (P<0.05). When GHR was blocked with neutralizing antibodies, this protective effect was eliminated. By contrast, thGH pre-incubation did not change the colony formation rate in GHR negative expression LoVo cells. GH rapidly induced Akt phosphorylation in HCT-8 cells by GH/GHR signaling, and this effect was blocked by PI-3 kinase inhibitor. Conclusions The protective effect of GH on colorectal cancer cells may occur after radiotherapy exposured by GHR, which is related to the reduction of apotosis.

Objective: To investigate the prevalence of hyperactivity behavior in children aged 3-6 in Yunnan Province, to explore its relationship with neuropsychological development, so as to provide clues for early prevention and intervention of attention deficit hyperactivity disorder (ADHD) in children. Methods: A total of 1 321 children aged 3 to 6 from 10 kindergartens in 5 prefectures (cities) of Yunnan Province were selected by stratified random sampling method from October 2022 to May 2023. Teacher Rating Scale (TRS) was used to investigate childrens hyperactive behavior and coexistent behavior. A qualified evaluator applied the Developmental Scale for Children Aged 0-6 Years to assess the development of 5 ability areas of gross motor movement，fine movement，adaptive ability，language and social behavior. Statistical analysis was performed using Wilcoxon rank sum test and χ2 test. Binary Logistic regression was applied to analyze the score of their hyperactivity behavior and its relationship with other behavior problems and neuropsychological development. Results: The detection rate of hyperactivity behavior was 8.6% in children aged 3 to 6 years, 12.8% in boys and 4.1% in girls (χ2=31.53, P<0.01). The detection rate of hyperactivity in 3yearold children was 13.9%, which was higher than that in 4yearold (9.2%) and 5yearold children (7.0%) (χ2=8.73, P<0.05). The detection rate of inattentionpassivity of rural children (14.6%) was higher than that of urban children (5.9%) (χ2=22.23, P<0.01). Binary Logistic regression analysis showed that the higher the level of adaptive development, the lower the risk of hyperactivity (OR=0.58, 95%CI=0.39-0.86), the higher the risk of hyperactivity (OR=0.57, 95%CI=0.35-0.91), the higher the risk of conduct problems (OR=0.57, 95%CI=0.37-0.87), inattentionpassivity (OR=0.49, 95%CI=0.33-0.74) were also at lower risk (P<0.05). Children with higher levels of fine motor development had a lower risk of inattentionpassivity (OR=0.59, 95%CI=0.37-0.93, P<0.05). Conclusions Hyperactivity in boys and inattentionpassivity in rural children requires more attention. It is necessary to strengthen childrens early adaptive ability and fine motor training to prevent hyperactive behavior and inattention.

OBJECTIVETo explore the feasibility of short-term neoadjuvant chemotherapy (NACT) in patients with advanced gastric cancer (AGC), and to compare clinical efficacy of short-term neoadjuvant chemotherapy with different ways.

METHODSClinical data of 310 AGC patients treated with one course of NACT using EOF regimen(epirubicin, oxaliplatin and fluorouracil plus calcium folinate) in our hospital from January 2008 to December 2011 were retrospectively analyzes. Efficacy was compared between regional arterial infusion chemotherapy and intravenously chemotherapy.

RESULTSAll the 310 AGC patients completed one course of NACT and none was interrupted by adverse events. Postoperative pathological remission rate was 33.9% (105/310) and 5 patients (1.6%) had complete pathological remission. The pathologic response rate in the regional arterial infusion chemotherapy group was higher than that in the intravenously chemotherapy group(42.4% vs. 23.6%, P = 0.001). Multivariate analysis revealed that chemotherapy method(HR=1.827, 95% CI:1.006-3.316, P = 0.048) was associated with significantly higher pathologic response.

CONCLUSIONSPathological response rate is quite low following short-term NACT. Regional arterial infusion chemotherapy with short-term NACT can improve the pathological response rate of advanced gastric cancer.

Antineoplastic Combined Chemotherapy Protocols ; administration & dosage ; therapeutic use ; Epirubicin ; Fluorouracil ; Humans ; Infusions, Intra-Arterial ; Leucovorin ; Neoadjuvant Therapy ; Organoplatinum Compounds ; Remission Induction ; Retrospective Studies ; Stomach Neoplasms ; drug therapy

Background/Aims: Acute-on-chronic liver failure (ACLF) is a catastrophic illness. Few studies investigated the prognostic value of vitamin D-binding protein (VDBP) for hepatitis B virus (HBV)-related ACLF (HBV-ACLF) resulted in conflicting results. Methods: Two prospective HBV-ACLF cohorts (n=287 and n=119) were enrolled to assess and validate the prognostic performance of VDBP. Results: VDBP levels in the non-survivors were significantly lower than in the survivors (P<0.001). Multivariate Cox regression demonstrated that VDBP was an independent prognostic factor for HBV-ACLF. The VDBP level at admission gradually decreased as the number of failed organs increased (P<0.001), and it was closely related to coagulation failure. The areas under the receiver operating characteristic curve (AUCs) of the Child-Pugh-VDBP and chronic liver failuresequential organ failure assessment (CLIF–SOFA)-VDBP scores were significantly higher than those of Child-Pugh (P<0.001) and CLIF-SOFA (P=0.0013). The AUCs of model for end-stage liver disease (MELD)-VDBP were significantly higher than those of MELD (P= 0.0384) only in the case of cirrhotic HBV-ACLF patients. Similar results were validated using an external multicenter HBV-ACLF cohort. By longitudinal observation, the VDBP levels gradually increased in survivors (P=0.026) and gradually decreased in non-survivors (P<0.001). Additionally, the VDBP levels were found to be significantly decreased in the deterioration group (P=0.012) and tended to be decreased in the fluctuation group (P=0.055). In contrast, they showed a significant increase in the improvement group (P=0.036). Conclusions The VDBP was a promising prognostic biomarker for HBV-ACLF. Sequential measurement of circulating VDBP shows value for the monitoring of ACLF progression.

Objective:To explore the significance of triggering receptor expressed on myeloid cells-2 (TREM-2) prognostic evaluation so as to provide novel biological markers in clinical practice for patients with hepatitis B virus-related acute-on-chronic liver failure ( HBV-ACLF).Methods:The research subjects of this study were divided into an experimental group and a control group. Fifty HBV-ACLF cases admitted to the Department of Infectious Diseases of the First Affiliated Hospital of Nanchang University from January 1, 2019 to December 31, 2019 were selected as the experimental group. Patients were divided into survival and death groups according to the actual prognosis at discharge (self-discharge and dead patients were considered death groups, and all enrolled patients were hospitalized for more than 28 days). Twenty-five healthy subjects were chosen as the control group. Peripheral venous blood was collected from the experimental group and the control group. Plasma and peripheral blood mononuclear cells (PBMC) were isolated. The concentrations of TREM-2, interleukin (IL)-6, and IL-8 were detected in the plasma. TREM-2 mRNA expression was detected in PBMC. A single blood sample was collected from the control group, whereas five blood samples were dynamically collected from the experimental group on the day of admittance and at 7, 14, 21, and 28 days after treatment commenced. Simultaneously, upon admission, the relevant clinical indicators of HBV-ACLF patients were monitored, including the liver function test: alanine aminotransferase, aspartate aminotransferase, total bilirubin, albumin, coagulation function test: international normalized ratio, prothrombin time, and other indicators. Measurement data were expressed as mean±standard deviation (x±s). Count data were compared and analyzed using the χ 2 test. The intra-group factor mean was compared using a repeated measures ANOVA. The means were analyzed by t-tests between the two groups. Bivariate correlation analysis was used to analyze the correlation between the two variables. The value of TREM-2 as a diagnostic marker was analyzed using the receiver operating characteristic (ROC) curve. Results:The mRNA expression of TREM-2 in the PBMC of HBV-ACLF patients showed a gradually increasing trend at various time points and was significantly higher in the survival group than that of the control group at 28 days ( P < 0.01), while the death group showed a gradually weakening trend at various time points and was significantly lower than the control group at 28 days ( P < 0.01). (1) The levels of TREM-2 in the plasma of HBV-ACLF patients generally showed a gradually increasing trend at various time points in the survival group. The levels on the day of admission and 7, 14, 21, and 28 days after the initiation of treatment were (1.49±0.85), (1.62±0.58), (1.95±0.69), (2.33±0.71), and (2.00±0.67) ng/ml, respectively. The expression of TREM-2 in the death group showed a gradually weakening trend at various time points. The levels on the day of admission and 7, 14, 21, and 28 days after initiation of treatment were (1.40±0.73), (1.59±0.79), (1.56±0.80), (1.05±0.49), and (0.81±0.21) ng/ml, respectively. The survival group's various detection time points were higher than those of the death group, and the difference was statistically significant. The plasma level of TREM-2 in the healthy control group was (1.25±0.35) ng/ml. (2) The concentrations of IL-6 and IL-8 in the plasma of HBV-ACLF patients showed a gradually decreasing trend at various time points in the survival group. The levels on the day of admission and 7, 14, 21, and 28 days after initiation of treatment were (46.70±26.31), (33.98±20.28), (19.07±10.24), (14.76±7.84), (9.12±7.65) and (108.29±47.07), (93.85±26.53), (79.27±34.63), (56.72 ±18.30), (37.81±13.88) pg/ml, respectively. However, its concentration in the death group fluctuated within a relatively high range. The levels on the day of admission and 7, 14, 21, and 28 days after the initiation of treatment were (41.94±24.19), (36.99±19.78), (34.30±20.62), (34.14±14.52), (36.64±23.61) and (104.65±50.16), (112.98±45.03), (118.43±45.00), (111.67±40.44), (109.55±27.54) pg/ml, respectively. (3) Bivariate correlation analysis results indicated that the plasma TREM-2 content was negatively correlated with the plasma levels of pro-inflammatory cytokines IL-6 and IL-8 ( r = -0.224, P = 0.025; r = - 0.223, P = 0.026). ROC curve analysis showed that the mRNA levels of TREM-2 in PBMCs at various time points for prognostic evaluation of HBV-ACLF patients were 1d=0.667, 7d=0.757, 14d=0.979, 21d=0.986, and 28d= 0.993. The areas under the ROC curve of the TREM-2 content in the plasma at various time points were 1d=0.522, 7d=0.571, 14d=0.658, 21d=0.927, and 28d=0.994. Conclusion:TREM-2 mRNA expression in PBMC and TREM-2 content in plasma have a significant relationship to the prognosis of HBV-ACLF patients and may inhibit the liver inflammatory response by regulating the secretion of pro-inflammatory cytokines IL-6 and IL-8. Dynamic monitoring of TREM-2 expression in peripheral blood is favorable for evaluating the prognostic condition of HBV-ACLF patients.

Objective:To investigate the present situation of unintended pregnancy within two years postpartum and its influencing factors in China.Methods:Participants who delivered a live birth at 60 hospitals in 15 provinces in the eastern, central and western regions of China during July 2015 to June 2016 were interviewed by using structured questionnaire. Information on occurrence of unintended pregnancy within 2 years after delivery, postpartum contraceptive use, sexual resumption, breastfeeding, and women′s socio-demographic characteristics, and so on, were collected. Life-table analysis, cluster log-rank tests and a 2-level Cox regression model were used for data analysis.Results:A total of 18 045 postpartum women were investigated. The cumulative 1- and 2-year unintended pregnancy rates after delivery were 5.3% (95% CI: 4.5%－6.1%) and 13.1% (95% CI: 11.3%－14.8%), respectively. Cox regression model analysis showed that the risk of unintended pregnancy within 2 years postpartum were increased in younger women, ethnic minorities, women with abortion history, and those who had a vaginal delivery with short lactation time and late postpartum contraceptive initiation (all P<0.01). The risk of postpartum unintended pregnancy was not associated with geographic regions and hospitals where women gave a birth (all P>0.05). Conclusions:In China, the risk of unintended pregnancy within 2 years after delivery is relatively high. Service institutions and service providers should improve the quality of postpartum family planning services, promote the use of high effect contraceptive methods, and educate women to use a method at the time of their sexual resumption or even before.

Objective: To investigate the expression and distribution of sclerostin in the alveolar bone of rat in the absence of estrogen, and to provide evidence for the analysis of the histological correlation between sclerostin and alveolar bone remodeling in rats. Methods : The experimental subjects of this study were 32 8-week-old female Wistar rats. Among them, 16 rats were ovariectomized (OVX), and 16 rats were subjected to a sham operation (Sham). These rats were sacrificed 1, 2, 3, and 4 weeks after the operation, and the mandibles were removed and embedded. The mesial and distal sections of the rat,s mandibular first molars were selected and stained with anti-tartrate phosphatase (TRAP), sclerostin immunostaining, multiple immunostainings, RANKL and TRAP double staining, and silver-plated multiple staining. Results : As the postoperative time in rats increased, the TRAP-positive osteoclasts counts in the OVX group in the interalveolar septum of mandibular first molar increased significantly, and statistical difference was noted between the groups (P ＜ 0.05). The OVX 2w, 3w, and 4w groups exhibited more TRAP-positive osteoclasts compared with the Sham group at the corresponding time point, and the results were statistically different (P ＜ 0.05). Sclerostin immunostaining revealed that the proportion of positive bone cells in the mesial side of the periodontal ligament area of mandibular first molar in the OVX group gradually decreased. Statistical differences were noted between the OVX 3w group and the OVX 4w group as well as the OVX 1w group and, the OVX 2w group (P ＜ 0.05). In the comparison between the area near the periodontal ligament and the central area of the alveolar bone septum of the mandibular first molar in the same group, the positive expression ratio of sclerostin in the OVX 3w and OVX 4w groups in the area near the periodontal ligament was reduced compared with that in the central area of the alveolar bone septum. The results were statistically significant (P ＜ 0.05). A larger number of osteoblasts was noted around the osteoclasts in the OVX 4w group compared with the Sham 4w group based on ALP/ TRAP /sclerostin multiple staining, whereas less sclerostin-positive osteoblasts were noted in the OVX 4w group. Sclerostin/TRAP/silver plating staining showed that the bone tubules around the sclerostin positive bone cells mostly exhibited a parallel and neat arrangement, and the bone tubules around sclerostin negative bone cells were more irregular and disorderly arranged in the OVX 4w group Conclusion Sclerostin protein is involved in alveolar bone remodeling in estrogen-deficient rats.