Objective To explore a novel and highly specific small-molecule TNFβinhibitor by using computer-aid?ed virtual screening and cell-based assays in vitro. Methods Computer-aided drug design and virtual screening were used to design and identify chemical compounds that targeted TNFβbased on the crystal structure of the TNFβ-TNFR1 com?plex. The effect of the small-molecule compound against TNFβ-induced cytotoxicity of L929 cells was detected by MTT as?say, and the efficacy of the compound to inhibit TNFβ-induced apoptosis of L929 cells was determined by flow cytometry as?say. The impact of the compound on L929 cell cycle was examined by Propidium Iodide (PI) staining and flow cytometry, and the influence of the compound on TNFβ-triggered signal pathway was analyzed by Western blot assay and Ultra VIEW VOX 3D Live Cell Imaging System. Results No.35 compound (named as C35 thereafter) could effectively inhibit TNFβ-induced cell death in a dose dependent manner, and the half-maximum inhibition concentration (IC50) was 8.19μmol/L. Furthermore, C35 had lower cytotoxicity and minimal effect on L929 proliferation. Here we further revealed that C35 could affect TNFβ-induced apoptotic pathway by blocking the activation of Caspase 3, and markedly reduce L929 cell apoptosis induced by TNFβ. Conclusion A novel TNFβsmall-molecule inhibitor was identified by combining computer-aided virtual screening with functional assays, and which could block TNFβ-triggered apoptotic pathway and efficiently inhibit the cell death in?duced by TNFβ.

Objective To systematically evaluate the efficacy,safety and recent curative effects of delta-shaped anastomosis in totally laparoscopic distal gastrectomy (ICBI) for gastric carcinoma.Methods Literatures in English and Chinese comparing ECBI and ICBI published up to November 2015 were searched from international and national online databases.Meta-analysis was conducted by Review Manager 5.3 soft ware.Results There are eleven studies involved,with 2020 gastric cancer patients,including,1 169 ECBI cases and 851 ICBI cases.Meta-analysis showed that there was no significant statistical differences between ECBI group and ICBI group in operative time,resection margin length,overall postoperative complications and anastomosis-related complications.(all P ＞ 0.05).When compared to ECBI,the estimated blood loss was significantly less in ICBI,and ICBI with more retrieved lymph nodes,faster recovery of gastrointestinal function,less use of painkiller and shorter postoperative hospital stay (all P ＜ 0.05).Conclusion ICBI is safe and feasible in treatment of gastric cancer,and has a good short-term effect.

Objective To investigate the clinical features, diagnosis, treatment and prognosis of muhisystemic invasive fungal diseases. Methods Twenty-one patients with multisystemic invasive fungal diseases who were hospitalized in department of infectious diseases from January 2001 to June 2008 were retrospectively reviewed. The pathogenic bacteria, involved organs, underlying diseases, clinical manifestations, treatments and prognoses of muhisystemic invasive fungal diseases were analyzed. Results Among 21 recruited cases, 17 had underlying diseases and 11 were treated with long-term immunosuppressive agents. The main pathogenic bacteria were Cryptococcus neoformans, Aspergillus and Candida parapsilosis. Lung and brain were involved in 16 cases (skin involve in 2 cases and lymph node involved in 1 case simultaneously), lung and lumbar involved in 2 cases, heart valves involved in 2 cases, and liver, spleen and bone marrow involved in 1 case. Eight cases were cured, 6 were improved and 7 died. Conclusions In this study, most of the 21 cases with multisystemic invasive fungal diseases are immunocompromised. The main pathogenic bacterium is Cryptococcus neoformans. The lung and brain are common organs involved. Prognosis is associated with early diagnosis and active anti-fungal treatment.

OBJECTIVETo study the expressions of Notch1-4 gene in human keloid-derived mesenchymal-like stem cells, and to explore the Notch signaling pathway's role in the formation of keloid.

METHODSKeloid samples were collected to harvest human keloid-derived mesenchymal-like stem cells through two-step enzymatic dissociation method. By flow cytometry, cell phenotype of primary and P3 generation were analyzed. By immunocytochemistry, the expressions of Oct4, vimentin and CK19 were examined. Keloid-derived mesenchymal-like stem cells were induced into osteoblasts in vitro and calcium deposition was detected by Alizarin red S stain. Realtime polymerase chain reaction (RT-PCR) was used to detect the expressions of Notch1-4 mRNA in keloid-derived mesenchymal-like stem cells.

RESULTSFlow cytometry showed that keloid-derived mesenchymal-like stem cells of primary and P3 generation highly expressed CD29, CD44, CD90 from the typical MSC phenotype marker, but they failed to express HSC phenotype markers, such as CD34 and CD45. The results of immunocytochemistry showed that Oct4 from pluripotent stem cell markers and vimentin from mesenchymal cell markers was positive and CK19 from epithelial cell markers was negative. After induced differentiation into osteoblasts in vitro after 21 day, calcium nodules could be seen clearly; Notch1-4 gene were expressed in keloid-derived mesenchymal-like stem cells through RT-PCR. The relative quantitative of Notch2, Notch3 gene were higher than Notch1, Notch4 gene (P < 0.05).

CONCLUSIONSThe expression difference of different subtypes from Notch gene in human keloid-derived mesenchymal-like stem ceils may be related to self-renewal, proliferation, differentiation, and participate in the formation of keloid.

Adolescent ; Cells, Cultured ; Child ; Female ; Humans ; Keloid ; metabolism ; pathology ; Male ; Mesenchymal Stromal Cells ; metabolism ; Receptors, Notch ; metabolism

Objective To study interleukin-23 (IL-23) levels in serum and dendritic cells of acute-on-chronic liver failure (ACLF) patients with chronic hepatitis B (CHB) and to explore its relationship with the prognosis.Methods Peripheral blood mononuclear cells and serum were collected from 40 ACLF patients with CHB (including survival group 27 cases and non-survival group 13 cases) and 26 healthy controls.Monocytes were induced to immature dendritic cell in vitro and TNF-α was added to induce dendritic cell maturation.IL-23 mRNA of dendritic cells was detected by real time polymerase chain reaction (PCR), and serum IL-23 level was measured by enzyme-linked immuno sorbent assay (ELISA).Differences among the parameters with normal distribution were compared using t test, those with non-normal distribution were compared using non-parametric Mann-Whitney U-test, and the relationship between two variables was assessed by Spearman′s rank correlation.Results International normalized rate (INR) and model for end-stage liver disense (MELD) scores in non-survival group of ACLF were higher than those in survival group (INR: 2.32 vs 1.64, U=69.00, P=0.002 2;MELD:36 vs 30, U=64.50, P=0.001 4).However, there were no significant differences between two groups at gender, age, alanin aminotransferase (ALT), aspartate aminotrans ferase (AST), bilirubin, creatinine, hepatitis B virus (HBV) DNA, hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg) and serum IL-23.IL-23 mRNA level in dendritic cells at baseline in non-survival group of ACLF was significantly higher than that in survival group (76 vs 43, U=71.50, P=0.002 8).After treatment, serum IL-23 was significantly declined in survival group ([160±75] ng/L vs [91±49] ng/L, t=4.012, P=0.000 2), but not in non-survival group.Significant positive correlation was observed between IL-23 mRNA level in dendritic cells and MELD score at baseline (r=0.7198,P<0.01).Conclusions Persistent high serum IL-23 level suggests poor prognosis in ACLF patients with CHB.IL-23 mRNA expression in dendritic cells has good consistency with MELD score and the patients with high IL-23 mRNA expression has poor outcome.

Objective:To investigate the clinical value of sevoflurane inhalation combined with epidural anesthesia in the operation of digestive tract malignant tumors in the elderly.Methods:From August 2016 to September 2017, 74 elderly patients with digestive tract malignant tumors who were operated in the Integrated Traditional Chinese and Western Medicine Hospital of Wenzhou were divided into two groups by random number table method, with 37 cases in each group.The control group was given sevoflurane inhalation anesthesia, while the observation group was given sevoflurane inhalation combined with epidural anesthesia.The changes of blood pressure, heart rate and cognitive function were observed.Results:There were no statistically significant differences in blood pressure and heart rate between the two groups at 5 minutes after tracheal intubation (all P>0.05). After 0 and 10 min of recovery of bilateral lung ventilation, the blood pressures of the observation group were (74.75±4.93)mmHg, (72.79±4.60)mmHg, respectively, which were lower than those of the control group [(82.44±3.62)mmHg, (75.70±5.14)mmHg] ( t=7.648, 2.562, all P<0.05). The heart rate of recovery of bilateral pulmonary ventilation in the observation group was (70.75±4.21) times/min, which in the control group was (75.91±3.89)times/min, the difference was statistically significant between the two groups( t=5.476, P<0.05). The cognitive function scores of the observation group were (25.74±0.69)points, (26.13±1.49)points, (27.90±0.77)points, (28.26±1.18)points, respectively, which were higher than those of the control group [(19.88±2.30)points, (22.85±0.96)points, (24.38±1.21)points, (24.35±1.63)points] ( t=14.844, 11.256, 14.929, 11.819, all P<0.05). Conclusion:Sevoflurane inhalation combined with epidural anesthesia is effective for elderly patients with digestive tract malignant tumors, which can avoid excessive fluctuation of physical signs and promote the recovery of cognitive function.

Objective:To investigate the effect of iguratimod (IGU) on transforming growth factor-β 1 (TGF-β 1)-induced primary human lung fibroblasts (pHLFs) activation and collagen secretion. Methods:Mice pulmonary fibrosis (PF) models were established in vivo and were divided into three groups: the control group (CTR group), the Bleomycin (BLM) group and the BLM+IGU group, hematoxylin-eosin (HE) staining was used to observe lung morphology, and Masson staining was used to observe the degree of collagen accumulation in lung. Fibronectin and smooth muscle 22 (SM22) were detected by immunofluorescence, and the content of hydroxyproline in lung tissue was detected by chloramine-T method. In vitro, pHLFs were used to assess the effect of IGU on TGF-β 1 stimulation in four groups: CTR group, IGU group, TGF-β 1 group and TGF-β 1+IGU group, the apoptosis of cells was detected by flow cytometry, and the mRNA expression of collagen type Ⅰ (COL-Ⅰ) and collagen type Ⅲ (COL-Ⅲ) was detected by quantitative real-time polymerase chain reaction (qRT-PCR). The protein levels of α-smooth muscle actin (α-SMA), fibronectin, p-Smad2, p-Smad3 and transcription coactivator p300 were detected by Western blot and immunofluorescence. One-way ANOVA was used for all data, and LSD- t test or Kruskal-Wallis test was used for pair comparison. Results:The content of hydroxyproline in CTR group, the BLM group and the BLM+IGU group was (0.552±0.075) μg/mg, (1.293±0.081) μg/mg and (0.833±0.053) μg/mg ( F=169.672, P<0.01) respectively. IGU reduced the content of hydroxyproline in the lung tissue of mice, reduced the accumulation of collagen in the lung, and thus reduced the degree of BLM-induced pulmonary fibrosis, and improved the pathological changes in the lung of mice. In cell experiments, IGU had no significant effect on apoptosis ( F=0.83, P=0.54). The relative expression levels of COL-Ⅰ mRNA in the CTR group, TGF-β 1 group and TGF-β 1+IGU group were (100.4±1.2), (299.0± 13.0) and (202.5±7.0) respectively ( F=468.7, P<0.01). The relative expression levels of COL-Ⅲ mRNA in the CTR group, TGF-β 1 group and TGF-β 1+IGU group were (99.8±1.9), (350.6±8.0) and (220.3±9.9) respectively ( F=468.7, P<0.01). The relative expression levels of α-SMA protein were (0.193±0.038) in CTR group, (0.530±0.061) in TGF-β 1 group, and (0.410±0.065) in TGF-β 1+IGU group ( F=35.620, P<0.01); The relative expression levels of fibronectin in CTR group, TGF-β 1 group, and TGF-β 1+IGU group were (0.200±0.020), (0.700±0.020) and (0.410±0.066) respectively ( F=123.326, P<0.01). The relative expression levels of p-Smad3 protein in CTR group, TGF-β 1 group, and TGF-β 1+IGU group were (0.120±0.020), (0.573±0.586) and (0.327±0.252) respectively( F=92.987, P<0.01); The relative expression levels of p300 in CTR group, TGF-β 1 group and TGF-β 1+IGU group were (0.180±0.055), (0.923±0.025) and (0.650±0.050) respectively ( F=207.676, P<0.01). IGU significantly decreased the mRNA expression levels of COL-Ⅰ and COL-Ⅲ induced by TGF-β 1, inhibited the protein expression levels of α-SMA, fibronectin, p300, and phosphorylation of Smad2/3. Conclusion:Our results revealed the beneficial effect of IGU on the inhibition of TGF-β 1-mediated pHLFs activation and collagen secretion via the Smad3/p300 pathway, thus suggest that it might act as an effective anti-fibrotic agent in preventing the progression of PF.

Objective:To explore the risk factors for 30-day death in emergency department patients, and then construct a prediction model and validate it using nomogram.Methods:A retrospective cohort study was conducted. The clinical data of 1 091 patients admitted to the emergency department of the First People's Hospital of Changde from January 1 to June 30, 2021 was collected, including 741 patients from January 1 to March 31 in the development group and 350 patients from April 1 to June 30 in the validation group. General information, first vital signs admitted to the emergency department, and laboratory results were collected, the modified early warning score (MEWS) was calculated, and 30-day outcomes were recorded. Univariate and multivariate Logistic regression analysis was used to screen out the risk factors of 30-day death. According to the results of multivariate analysis, the nomogram was used to construct a 30-day death prediction model. The receiver operator characteristic curve (ROC curve) was used to evaluate the consistency of the prediction model, the calibration of the prediction model was evaluated by the Hosmer-Lemeshow goodness of fit test.Results:A total of 1 091 patients were enrolled. There were 741 patients in the development group, including 356 males and 385 females, aged (51.42±17.33) years old, and the 30-day mortality was 28.88%. There were 350 patients in the validation group, including 188 males and 162 females, aged (52.88±16.11) years old, and the 30-day mortality was 24.00%. The results of the univariate analysis showed that age, primary diagnosis on admission, consciousness, respiratory rate (RR), systolic blood pressure (SBP), heart rate (HR), pulse oxygen saturation (SpO 2), MEWS score, erythrocyte sedimentation rate (ESR), procalcitonin (PCT) and body mass index (BMI) might be the risk factors for 30-day death in patients in the emergency department. The results of the multivariate analysis showed that the MEWS score [odds ratio ( OR) = 14.22, 95% confidence interval (95% CI) was 1.46-138.12], ESR ( OR = 46.71, 95% CI was 20.48-106.53), PCT ( OR = 4.97, 95% CI was 2.46-10.02), BMI (24.0-27.9 kg/m 2: OR = 37.82, 95% CI was 14.69-97.36; ≥28.0 kg/m 2: OR = 62.11, 95% CI was 25.77-149.72) were independent risk factors for 30-day death in the emergency department (all P < 0.05). Using the four variables with the results of multivariate analysis to construct a nomogram prediction model, the area under the ROC curve (AUC) was 0.974 (95% CI was 0.753-0.983) for the development group, and the AUC was 0.963 (95% CI was 0.740-0.975) for the validation group. The Hosmer-Lemeshow test showed no statistically significant difference between the predicted outcome of the nomogram prediction model and the actual occurrence ( χ2 = 1.216, P = 1.270). Conclusion:The prediction model developed by the MEWS score combined with BMI, ESR and PCT can scientifically and effectively predict the 30-day outcome of emergency department patients.

OBJECTIVE To determine the genetic cause of a child with blepharophimosis, ptosis, and epicanthus inverses syndrome and tetralogy of Fallot, and to correlate the phenotype with the genotype. METHODS Routine G-banding has been previously performed on the patient and her parents. Chromosome microarray analysis (CMA) was performed for the three individuals and the fetus. RESULTS Chromosomal analysis has suggested normal karyotypes for the child and her parents. However, a de novo 8.9 Mb deletion on chromosome 3q22.1-q23 was detected by CMA. The deleted region has encompassed 74 genes including 41 disease-related genes, and this is also the most frequent region involved in interstitial 3q deletion. Patients with deletion of this region often have a common feature of dysplasia of eyelids, as well as a spectrum of other anomalies according to different breakpoints, including microcephaly, skeletal anomalies, congenital heart defects, cranial anomalies, intellectual disability and developmental delay. The patient's phenotype was in accordance with such spectrum. Her parents and sib did not show this variation by CMA. CONCLUSION The de novo interstitial deletion of 3q22.1-q23 probably underlies the main clinical manifestation in this child. CMA can provide more detailed information and allow further investigation of the genotype-phenotype correlation.
Blepharophimosis ; genetics ; Child, Preschool ; Chromosomes, Human, Pair 3 ; Female ; Humans ; Mitochondrial Proteins ; genetics ; Phenotype ; Ribosomal Proteins ; genetics ; Skin Abnormalities ; genetics ; Tetralogy of Fallot ; genetics ; Urogenital Abnormalities ; genetics

BACKGROUND/AIMS: To investigate the role of selected serum microRNA (miRNA) levels as potential noninvasive biomarkers for differentiating S0–S2 (early fibrosis) from S3–S4 (late fibrosis) in patients with a chronic hepatitis B virus (HBV) infection. METHODS: One hundred twenty-three treatment-naive patients with a chronic HBV infection who underwent a liver biopsy were enrolled in this study. The levels of selected miRNAs were measured using a real-time quantitative polymerase chain reaction assay. A logistic regression analysis was performed to assess factors associated with fibrosis progression. Receiver operating characteristic (ROC) curve and discriminant analyses validated these the ability of these predicted variables to discriminate S0–S2 from S3–S4. RESULTS: Serum miR-29, miR-143, miR-223, miR-21, and miR-374 levels were significantly downregulated as fibrosis progressed from S0–S2 to S3–S4 (p < 0.05), but not miR-16. The multivariate logistic regression analysis identified a panel of three miRNAs and platelets that were associated with a high diagnostic accuracy in discriminating S0–S2 from S3–S4, with an area under the curve of 0.936. CONCLUSIONS: The levels of the studied miRNAs, with the exception of miR-16, varied with fibrosis progression. A panel was identified that was capable of discriminating S0–S2 from S3–S4, indicating that serum miRNA levels could serve as a potential noninvasive biomarker of fibrosis progression.
Biomarkers ; Biopsy ; Early Diagnosis* ; Fibrosis ; Hepatitis B virus ; Hepatitis B* ; Hepatitis B, Chronic ; Hepatitis* ; Humans ; Liver Cirrhosis* ; Liver* ; Logistic Models ; MicroRNAs* ; Polymerase Chain Reaction ; ROC Curve