6-Shogaol and 10-Shogaol Synergize Curcumin in Ameliorating Proinflammatory Mediators via the Modulation of TLR4/TRAF6/ MAPK and NFκB Translocation

Extensive research supported the therapeutic potential of curcumin, a naturally occurring compound, as a promising cytokinesuppressive anti-inflammatory drug. This study aimed to investigate the synergistic anti-inflammatory and anti-cytokine activities by combining 6-shogaol and 10-shogaol to curcumin, and associated mechanisms in modulating lipopolysaccharides and interferon-ɣ-induced proinflammatory signaling pathways. Our results showed that the combination of 6-shogaol-10-shogaol-curcumin synergistically reduced the production of nitric oxide, inducible nitric oxide synthase, tumor necrosis factor and interlukin-6 in lipopolysaccharides and interferon-γ-induced RAW 264.7 and THP-1 cells assessed by the combination index model. 6-shogaol-10-shogaol-curcumin also showed greater inhibition of cytokine profiling compared to that of 6-shogaol-10-shogaol or curcumin alone. The synergistic anti-inflammatory activity was associated with supressed NFκB translocation and downregulated TLR4-TRAF6-MAPK signaling pathway. In addition, SC also inhibited microRNA-155 expression which may be relevant to the inhibited NFκB translocation. Although 6-shogaol-10-shogaol-curcumin synergistically increased Nrf2 activity, the anti-inflammatory mechanism appeared to be independent from the induction of Nrf2. 6-shogaol-10-shogaol-curcumin provides a more potent therapeutic agent than curcumin alone in synergistically inhibiting lipopolysaccharides and interferon-γ induced proinflammatory mediators and cytokine array in macrophages. The action was mediated by the downregulation of TLR4/TRAF6/MAPK pathway and NFκB translocation.

Application of two noninvasive scores in predicting the risk of respiratory failure in full-term neonates: a comparative analysis / 中国当代儿科杂志

OBJECTIVES@#To study the value of Silverman-Anderson score versus Downes score in predicting respiratory failure in full-term neonates.@*METHODS@#The convenience sampling method was used to select the full-term neonates with lung diseases who were hospitalized in the neonatal intensive care unit from July 2020 to July 2021. According to the diagnostic criteria for neonatal respiratory failure, they were divided into a respiratory failure group (65 neonates) and a non-respiratory failure group (363 neonates). Silverman-Anderson score and Downes score were used for evaluation. The receiver operating characteristic analysis was used to compare the value of the two noninvasive scores in predicting respiratory failure in full-term neonates.@*RESULTS@#Among the 428 full-term neonates, 65 (15.2%) had respiratory failure. The Silverman-Anderson score had a significantly shorter average time spent on evaluation than the Downes score [(90±8) seconds vs (150±13) seconds; P<0.001]. The respiratory failure group had significantly higher points in both the Silverman-Anderson and Downes scores than the non-respiratory failure group (P<0.001). The Silverman-Anderson score had an AUC of 0.876 for predicting respiratory failure, with a sensitivity of 0.908, a specificity of 0.694, and a Youden index of 0.602 at the optimal cut-off value of 4.50 points. The Downes score had an AUC of 0.918 for predicting respiratory failure, with a sensitivity of 0.723, a specificity of 0.953, and a Youden index of 0.676 at the optimal cut-off value of 6.00 points. The Downes score had significantly higher AUC for predicting respiratory failure than the Silverman-Anderson score (P=0.026).@*CONCLUSIONS@#Both Silverman-Anderson and Downes scores can predict the risk of respiratory failure in full-term neonates. The Silverman-Anderson score requires a shorter time for evaluation, while the Downes score has higher prediction efficiency. It is recommended to use Downes score with higher prediction efficiency in general evaluation, and the Silverman-Anderson score requiring a shorter time for evaluation can be used in emergency.

Clinical characteristics and prognosis of central nervous system tuberculosis in adults / 中华传染病杂志

Objective:To investigate the clinical characteristics of central nervous system tuberculosis in adults and the possible factors affecting the mortality and disability of the patients.Methods:The clinical data of patients diagnosed as "tuberculous meningitis" "tuberculous meningoencephalitis" "tuberculous cerebrospinal meningitis" or "tuberculous brain ubscess" in Huashan Hospital, Fudan University and Jing′an Branch, Huashan Hospital, Fudan University in Shanghai from January 1, 2010 to December 31, 2017 were collected, and a retrospective cohort was established. The clinical characteristics were analyzed, Medical Research Council (MRC) grade system was used to assess the severity of meningitis, and the modified Rankin Scale was used to assess the impairment of self-care. Survival rate and disability rate of the cohort were analyzed. Binary logistic regression was used for multivariate analysis. Kaplan-Meier survival curve was used for survival analysis.Results:A total of 161 patients with central nervous system tuberculosis were enrolled. Among the 161 patients, 55 cases (34.2%) were confirmed, 72 cases (44.7%) were highly suspected and 34 cases (21.1%) were suspected diagnosis. There were 56 cases (34.8%) with MRC grade Ⅰ, 76 cases (47.2%) with MRC grade Ⅱ and 29 cases (18.0%) patients with MRC grade Ⅲ before treatment. Up to January 1, 2019, ten (6.2%) patients died, 32 (19.9%) patients lost to follow-up, 119 (73.9%) patients survived. The five-year survival rate was 92.83%. There were 72 patients with no impact on life, 34 patients with moderate impact and 13 patients with severe impact. The disability rate was 39.5% (47/119). Binary logistic regression analysis showed that increasing age (odds ratio ( OR)=1.06, 95%confidence interval ( CI) 1.00 to 1.13, P=0.032) and deterioration of MRC grade during anti-tuberculosis treatment ( OR=89.00, 95% CI4.46 to 1 779.00, P=0.003) were independent risk factors for death. When severe disability and death were used as adverse outcomes, logistic regression analysis showed increasing age ( OR=1.07, 95% CI 1.01 to 1.13, P=0.035) and deterioration of MRC grade during anti-tuberculosis treatment ( OR=77.17, 95% CI4.45 to 1 337.00, P=0.003) were still independent risk factors for adverse outcomes. Conclusions:The mortality of central nervous system tuberculosis in adults in this cohort is relatively low, but the disability rate is still high. Increasing age and deterioration of MRC grade during anti-tuberculosis treatment are independent risk factors for death and disability.

Study on the effect of miR-223-3p regulating transcription factor Rbpj on the differentiation of Th1 and Th17 cells in rats with experimental autoimmune uveitis / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the regulatory effect of miR-223-3p on the expression of transcription factor Rbpj and on the differentiation of Th1 and Th17 cells in experimental autoimmune uveitis(EAU)rats.<p>METHODS: The regulatory role of miR-223-3p in Rbpj gene expression was investigated by a dual luciferase expression reporter system. In the present study, 24 female Lewis rats were randomly divided into EAU model group, normal control(NC)group and blank control(BC)group, and each group included 8 rats. The EAU model group was injected with interphotoreceptor retinoid-binding protein(IRBP)emulsion containing Mycobacterium tuberculin H37RA and complete Freund's adjuvant to induce uveitis, while the NC group was injected with an equal volume of emulsion without IRBP peptide. The rats in the BC group received the same volume of sterile saline solution. At 12d after immunization, the spleen, lymph node and eye tissues in both groups were aseptically isolated, and the expression levels of miR-223-3p and Rbpj RNAs were detected by real-time quantitative PCR(Q-PCR); Meanwhile, the expression levels of Rbpj, IFN-γ and IL-17 proteins were detected by ELISA, and the levels of Th1 and Th17 cell lineages in each tissue from each groups were detected by flow cytometry. <p>RESULTS: The results of dual fluorescein assay indicated that Rbpj was the target gene which regulated by miR-223-3p. At 12d after immunization, compared with NC group, the relative expression levels of miR-223-3p in spleen, lymph node and eye tissues from EAU model rats were 0.33±0.29, 0.11±0.12 and 0.18±0.11, respectively, accompanied by the down-regulated expression, and the differences were statistically significant(all <i>P</i><0.05); Rbpj mRNA levels were 3.00±0.06, 1.52±0.12 and 3.01±0.34, respectively, and were all up-regulated, while the differences were statistically significant(all <i>P</i><0.05). Moreover, the differences in miR-223-3p and Rbpj mRNA levels in spleen, lymph node and eye tissues of rats in the blank control group were not statistically significant compared with those in the NC group(<i>P</i>>0.05); ELISA results revealed that the expression levels of RBPJ, IFN-γ and IL-17 proteins in all tissues from EAU rats at 12d after immunization were significantly higher than those in the NC group( all <i>P</i><0.05), and there was no statistically significant difference in the expression levels of Rbpj, IFN-γ and IL-17 protein in all tissues of rats in the blank control group compared with the NC group(<i>P</i>>0.05); Meanwhile, flow cytometry results showed that the proportions of Th1 and Th17 cell lineages in all tissues from EAU model group were significantly higher than those from the NC group at 12d after immunization, and the differences were statistically significant(all <i>P</i><0.05). Furthermore,there was no significant change in the proportion of Th1 and Th17 cells in each tissue in the BC and NC groups(all <i>P</i> >0.05). <p>CONCLUSION: The miR-223-3p can negatively regulate the expression of the transcription factor Rbpj of Notch signaling pathway. The down-regulated miR-223-3p expression in EAU rats can increase the expression levels of Rbpj gene and protein, and aggravate the differentiation of Th1 and Th17 cells and the expression levels of related molecules IFN-γ and IL-17, which in turn affect the development of uveitis.

Effect of acupuncture on intestinal flora and TLR4 in brain and intestinal tissues in rats with stress gastric ulcer / 中国针灸

OBJECTIVE@#To observe the effect of acupuncture at "Guanyuan" (CV 4) and "Xiajuxu" (ST 39) on intestinal flora and Toll-like receptors-4 (TLR4) in brain and intestinal tissue in rats with stress gastric ulcer (SGU), and to explore the possible mechanism of acupuncture for SGU.@*METHODS@#Thirty-one male SD rats were randomly divided into a blank group (@*RESULTS@#Compared with the blank group, the gastric mucosal damage index was significantly increased in the model group (@*CONCLUSION@#Acupuncture at "Guanyuan" (CV 4) and "Xiajuxu" (ST 39) could alleviate SGU in rats, and its mechanism may be related to increasing the diversity of intestinal flora, promoting the disorder of intestinal flora to normal, and reducing the overexpression of TLR4 in brain and intestinal tissues.

Research advances in the regulatory role of Notch signaling pathway in the occurrence and development of eye diseases / 国际眼科杂志(Guoji Yanke Zazhi)

@#The Notch signaling pathway is an evolutionarily highly conserved signaling pathway that exists in a wide variety of tissue cells and plays an important role in embryonic development, cell proliferation, differentiation and apoptosis, as well as in the maintenance of multicellular biosynthesis, angiogenesis, and the development of various cancers. In recent years, it has been found that the Notch signaling pathway is abnormally activated in cataract, uveitis and other ophthalmic diseases, regulating the process of ocular disease development. This article provides a brief review of the regulatory role of the Notch signaling pathway in the development of ophthalmic diseases.

Influence of portal vein thrombosis on the prognosis of patients with liver cirrhosis based on propensity score matching / 临床肝胆病杂志

ObjectiveTo investigate the influence of portal vein thrombosis (PVT) on the short-term prognosis of patients with liver cirrhosis and the risk factors for the prognosis of patients with liver cirrhosis. MethodsA retrospective analysis was performed for the clinical data of the patients with liver cirrhosis who were hospitalized in our hospital from September 2018 to March 2020, among whom 58 patients with PVT were enrolled as PVT group and 116 patients without PVT were enrolled as non-PVT group, and 44 patients were selected from each group based on propensity score matching (PSM) at a ratio of 1∶1 to balance the covariates between groups. The t-test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test or the Fisher’s exact test was used for comparison of categorical data between two groups. The Kaplan-Meier method and the log-rank method were used to analyze survival status and bleeding before and after PSM, and the Cox risk model was used to analyze the risk factors for the prognosis of patients with liver cirrhosis. ResultsBefore PSM, the non-PVT group had a significantly higher overall survival rate than the PVT group (P=0.008), while after PSM, there was no significant difference in overall survival rate between the two groups (P=0.076). Before PSM, the non-PVT group had significantly lower incidence rates of upper gastrointestinal bleeding or rebleeding than the PVT group before and after PSM (P＜0.001), and the results after PSM were consistent with those before PSM (P=0.028). The multivariate analysis of the prognosis of the patients with liver cirrhosis before PSM showed that PVT (hazard ratio ［HR］=2.944, 95% confidence interval ［CI］: 1.364-6.441, P=0.007) and Model for End-Stage Liver Disease (MELD) score ≥15 (HR=3.531, 95% CI: 1.630-7.650, P=0.001) were risk factors for short-term death of the patients with liver cirrhosis, and the multivariate analysis after PSM showed that MELD score ≥15 (HR=3.312, 95% CI: 1.049-10457, P=0.041) was a risk factor for short-term death of the patients with liver cirrhosis. ConclusionLiver cirrhosis with PVT increases the risk of upper gastrointestinal bleeding or rebleeding, but it is not an independent risk factor for short-term death in patients with liver cirrhosis. MELD score ≥15 is an independent risk factor for short-term death in patients with liver cirrhosis.

Overexpression of mitofusion 2 inhibits acute respiratory distress syndrome pulmonary fibrosis and its mechanism / 中华危重病急救医学

Objective:To study the inhibitory effect of overexpression of mitofusion 2 (Mfn2) protein on acute respiratory distress syndrome (ARDS) pulmonary fibrosis and its mechanism.Methods:Human embryo lung fibroblasts (HELF) were cultured in vitro, and digested and passaged when the adherent rate of HELF reached 80%, and then the cells in good condition were selected for experiment. The ARDS cell model was reproduced by 5 mg/L of lipopolysaccharide (LPS, LPS group); 75 mol/L adenovirus vector carrying mitofusion 2 (Adv-Mfn2) was transfected into HELF (Adv-Mfn2+LPS group); at the same time, blank control group (complete medium culture) and Adv-vector+LPS group were set as controls. The cell proliferation was observed by sulforhodamine B (SRB) method at 0, 12, 24, 36 and 48 hours. After Hoechst 33342 staining, the morphological changes were observed under confocal microscope. Western blotting was used to detect the protein expressions of Bcl-2 and caspase-3. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect the gene expressions of Bcl-2 and caspase-3. Results:After LPS stimulation for 12-48 hours, the cell proliferation rates in the LPS group increased gradually, which were significantly higher than those in the blank control group [12 hours: (10.75±1.51)% vs. (0.73±1.22)%, 24 hours: (20.09±1.71)% vs. (1.15±1.12)%, 36 hours: (20.58±1.55)% vs. (1.20±1.12)%, 48 hours: (21.30±1.51)% vs. (1.23±1.10)%, all P < 0.01]. There was no statistically significant difference in the cell proliferation rate between the LPS group and the Adv-vector+LPS group. After overexpression of Mfn2, the cell proliferation rates at 12, 24, 36, 48 hours in the Adv-Mfn2+LPS group were (8.93±1.14)%, (10.52±1.24)%, (10.72±1.30)%, and (10.91±1.20)%, which were significantly lower than those in the LPS group (all P < 0.05). Confocal microscopy showed that some cells in the blank control group had nuclei of different sizes, and some nuclei fragmented or shrank to form apoptotic bodies. The nuclei of the cells in the LPS and Adv-vector+LPS groups were round or oval in size, and only a few apoptotic cells appeared. When Mfn2 was overexpressed, there were more apoptotic cells in the visual field in the Adv-Mfn2+LPS group than LPS group. Western blotting and RT-qPCR results showed that Bcl-2 expressions increased significantly after LPS stimulation in the LPS group as compared with the blank control group [Bcl-2 protein (Bcl-2/GAPDH): 0.68±0.01 vs. 0.29±0.01, Bcl-2 mRNA (2 -ΔΔCT): 2.23±0.34 vs. 1.00±0.00, both P < 0.01], and caspase-3 expressions decreased significantly [caspase-3 protein (caspase-3/GAPDH): 0.37±0.02 vs. 0.66±0.02, caspase-3 mRNA (2 -ΔΔCT): 0.31±0.05 vs. 1.00±0.00, both P < 0.01]. Compared with LPS group, the expressions of Bcl-2 after overexpression of Mfn2 in the Adv-Mfn2+LPS group were down-regulated [Bcl-2 protein (Bcl-2/GAPDH): 0.46±0.01 vs. 0.68±0.01, Bcl-2 mRNA (2 -ΔΔCT): 1.45±0.14 vs. 2.23±0.34, both P < 0.01], and the expressions of caspase-3 were up-regulated [caspase-3 protein (caspase-3/GAPDH): 0.54±0.02 vs. 0.37±0.02, caspase-3 mRNA (2 -ΔΔCT): 0.88±0.10 vs. 0.31±0.05, both P < 0.01]. Conclusion:Mfn2 protein is involved in ARDS pulmonary fibrosis, which may be related to mitochondrial mediated inhibition of cell proliferation.

Clinical and imaging features of middle ear hairy polyps / 中华耳鼻咽喉头颈外科杂志

Objective: To collect the clinical cases of middle ear hairy polyp, and to summarize the imaging features. Methods: We retrospectively analyzed the clinical data of four cases middle ear hairy polyp confirmed by surgical and pathologic between January 2007 and January 2020 at the Affiliated Eye & ENT Hospital of Fudan University. There were three females, one male, with two left ears and two right ears, aged from 1 to 59 years. The CT and MRI imaging of the patients, and the corresponding clinical manifestations were analyzed. Results: Hairy polyps originated from tympanum in one case, originated from Eustachian tube in two cases, exhibiting recurrent otorrhea without evident inducement. The other case, hairy polyps originated from the Eustachian tube pharyngeal orifice and protruded into the nasopharyngeal cavity, with pharynx discomfort and aural fullness, endoscope showed offwhite polypoid mass with a little hair. All the four cases presented polypoid soft tissue masses on CT and MRI imaging, containing soft tissue wall and a large amount of adipose tissue, with soft tissue in the center of the mass which liked the core, and enhanced. MRI showed stratified arrangement of fat and soft tissue in the wall of the mass. Four cases all had surgical treatment, postoperative pathology examination presented that hair follicles, mature sebaceous glands and other skin appendages were found under squamous epithelium. A large amount of adipose tissue, part of muscle tissue, cartilage tissue, and some fibro-collagenous tissue were proliferated in the mass, accompanied by collagen degeneration. Conclusion: The middle ear hairy polyps has imaging characteristics, the polypoid soft tissue mass usually looks smooth and contains a large amount of adipose tissue, with a soft tissue in the center, and can be suggestively diagnosed by CT and MRI.

Influence of metabolic syndrome on patients with recurrence of hypertriglyceridemic acute pancreatitis / 临床肝胆病杂志

ObjectiveTo investigate the features of patients with recurrence of hypertriglyceridemic acute pancreatitis (HTG-AP) and the influence of metabolic syndrome (MetS) on recurrence through a retrospective analysis. MethodsA retrospective analysis was performed for 132 patients with recurrent HTG-AP who were admitted to The Affiliated Hospital of Southwest Medical University from June 2013 to December(recurrence group), and 132 patients with non-recurrent HTG-AP who were treated during the same period of time were enrolled as control group. Related data were collected, including basic information (age, sex, diabetes, hypertension, and body mass index), clinical data (etiology, routine blood test results, and biochemical results), and complications ［systemic inflammatory response syndrome (SIRS), acute peripancreatic fluid collection (APFC), acute necrosis collection (ANC), acute respiratory distress syndrome (ARDS), acute renal failure (ARF), multiple organ dysfunction syndrome (MODS), and ICU admission rate］. The t-test was used for comparison of normally distributed continuous data between groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups; the chi-square test or the Fisher’s exact test was used for comparison of categorical data between groups. A logistic regression analysis was used for comparison of related variables, and the receiver operating characteristic (ROC) curve was used to validate the diagnostic value of variables. ResultsCompared with the control group, the recurrence group had a significantly higher proportion of patients with MetS, obesity, a high-density lipoprotein cholesterol (HDL-C) level of ＜1 mmol/L, or ANC and significantly higher levels of blood glucose and C-reactive protein (CRP) and white blood cell count (WBC) (χ2=9.138, χ2=6.823, χ2=13.251, χ2=9587, Z=-2.255, Z=-2.202, Z=-4.674, all P＜0.05). The 132 patients in the recurrence group were divided into MetS group with 92 patients and non-MetS group with 40 patients, and compared with the non-MetS group, the MetS group had significantly higher WBC and blood glucose and a significantly higher proportion of patients with SIRS, APFC, ANC, or progression to moderate-to-severe AP (MSAP) (t=2.434, t=4.881, χ2=4.513, χ2=7.936, χ2=7.031, χ2=7.150, all P＜0.05). MetS (odds ratio ［OR］=2.007, 95% confidence interval ［CI］: 1.152-3.497, P＜0.05), ANC (OR=2.270, 95%CI: 1.185-4.347, P＜0.05), and WBC (OR=1.135, 95%CI: 1.059-1.217, P＜0.05) were independent risk factors for recurrence in HTG-AP patients. The combination of MetS, ANC, and WBC had a relatively high value in predicting the recurrence of HTG-AP, with an area under the ROC curve of 0.692. In the recurrence group, the levels of triglyceride, blood glucose, and CRP and the proportion of patients with smoking, HDL-C ＜1 mmol/L, or obesity in 125 patients (7 patients were excluded due to incomplete case data) at the time of recurrence were significantly lower than those in 132 patients at initial admission (Z=-3.270,Z=-3.546,Z=-4.382,χ2=0547,χ2=8.259,χ2=5.143, all P＜0.05), with further reductions in the proportion of patients with MSAP, APFC, or ANC (χ2=4.086,χ2=11.930,χ2=4.967,all P＜005). ConclusionMetS aggravates disease severity in patients with recurrent HTG-AP and is an important factor for recurrence. Smoking cessation, weight loss, and oral lipid-lowering drugs can reduce disease severity at the time of readmission.

Effect of acupuncture on intestinal flora in rats with stress gastric ulcer / 中国针灸

OBJECTIVE@#To observe the effect of acupuncture at "Baihui" (GV 20), "Zhongwan" (CV 12) and "Zusanli" (ST 36) on intestinal flora in rats with stress gastric ulcer (SGU) , and to explore the mechanism of acupuncture promoting SGU recovery.@*METHODS@#Thirty-one SPF SD rats were randomly divided into a control group (7 rats), a model control group (8 rats), an acupuncture group (8 rats) and a medication group (8 rats). The rats in the model group, acupuncture group and medication group were selected to applied the improved restraint water-immersion stress method to establish the SGU model. After modeling, the rats in the control group and model group were fixed and restrained for 20 min every day for a total of 5 days; the rats in the acupuncture group were intervented with acupuncture at "Baihui" (GV 20), "Zhongwan" (CV 12) and "Zusanli" (ST 36), once a day, 20 min each time, and twisting needle for 30 s every 5 min for a total of 5 days; the rats in the medication group were gavaged by solution of omeprazole enteric-coated tablet (200 mg/mL), 2 mL for each rat, once a day. Guth method was used to calculate the gastric mucosal damage index (GMDI), HE staining was used to observe the pathological changes of gastric mucosa, and 16SrDNA identification was used to detect the structural abundance of intestinal flora.@*RESULTS@#Compared with the control group, the GMDI of rats in the model group was increased (<0.01), the gastric mucosal pathological changes were significant, and the intestinal flora richness index Chao1, Observed species and diversity index Shannon were all decreased (<0.05), the diversity index Simpson was increased (<0.05). Compared with the model group, the GMDI of rats in the acupuncture group and medication group was reduced (<0.01, <0.05), the gastric mucosal damage degree was reduced, and the intestinal flora richness index Chao1, Observed species and diversity index Shannon were all increased (<0.05) and the diversity index Simpson decreased (<0.05). Compared with the medication group, the GMDI of rats in the acupuncture group was reduced (<0.01), the recovery of gastric mucosal injury was better than that of the medication group.@*CONCLUSION@#Acupuncture can effectively improve gastric mucosal injury of SGU, and the mechanism may be related to increasing the diversity of intestinal flora and promoting the correction of the disordered intestinal flora.

An investigation into the mechanisms underlying the regulatory effect of the E2F6 transcription factor on proliferation and metastasis of malignant melanoma cells through β-catenin signaling pathway / 中华皮肤科杂志

Objective:To determine the expression of the E2F6 transcription factor in human malignant melanoma tissues and cell lines, and to evaluate the effect of E2F6 on proliferation, migration and invasion of a malignant melanoma cell line A375.Methods:Frozen tissues and paraffin-embedded tissue sections were collected from 50 cases of cutaneous malignant melanoma and 30 cases of pigmented nevus in Department of Dermatology, Chongqing Traditional Chinese Medicine Hospital from January 2012 to December 2017. Quantitative reverse transcription-PCR (qRT-PCR) was performed to determine the mRNA expression of E2F6 in the malignant melanoma and pigmented nevus tissues, as well as in 7 malignant melanoma cell lines (HM, A375, WM451, WM35, SK-MEL-1, Hs-695T and MDA-MB-435s) and pigmented nevus cells, and immunohistochemical study and Western blot analysis were conducted to determine the protein expression of E2F6 and β-catenin in the malignant melanoma tissues. An E2F6-inhibiting plasmid and a control plasmid were separately transfected into A375 cells by using a liposome-mediated transfection method, and the E2F6 gene-knockdown efficiency was verified by qRT-PCR and Western blot analysis. Cell counting kit-8 (CCK8) assay, soft-agar plate cloning assay, Transwell migration and invasion assays and 3D cell culture assay were conducted to evaluate the effect of E2F6 gene knockdown on the proliferation, migration and invasion of A375 cells, flow cytometry was performed to detect the cell cycle and apoptosis rate, and Western blot analysis was conducted to determine the protein expression of total β-catenin, activated β-catenin, c-Myc and cyclin D1. The comparison between two groups was carried out by t test, the comparison among several groups by one-way analysis of variance, and multiple comparisons by least significant difference t test; Pearson correlation coefficient was used to analyze the correlation between E2F6 and β-catenin expression in cutaneous malignant melanoma. Results:The E2F6 mRNA expression was significantly higher in the 7 malignant melanoma cell lines than in the pigmented nevus cells (all P < 0.001). qRT-PCR showed that the relative mRNA expression of E2F6 was significantly higher in the cutaneous malignant melanoma tissues (0.000 55 ± 0.000 17) than in the pigmented nevus tissues (0.000 18 ± 0.000 09, t = 3.22, P < 0.001). Both the immunohistochemical study and Western blot analysis showed significantly increased E2F6 protein expression, but decreased β-catenin protein expression in the cutaneous malignant melanoma tissues compared with the pigmented nevus tissues (all P < 0.001). Correlation analysis showed that E2F6 protein expression was negatively correlated with β-catenin expression in the malignant melanoma tissues (immunohistochemical study: r = -0.56, Western blot analysis: r = -0.63, both P < 0.01). After knockdown of the E2F6 gene in A375 cells, the mRNA and protein expression of E2F6 was significantly lower in the E2F6 inhibition group than in the control group ( t = 3.38, 2.76 respectively, both P < 0.001). CCK8 assay showed that the cellular proliferative ability was significantly lower in the E2F6 inhibition group than in the control group ( t = 4.58, P < 0.01) 48 hours after transfection; soft-agar plate cloning assay showed that the colony-formation ratio was significantly lower in the E2F6 inhibition group than in the control group ( t = 2.26, P < 0.001) ; Transwell migration and invasion assays showed that the number of cells crossing the chamber was significantly lower in the E2F6 inhibition group (165 ± 23, 96 ± 11 respectively) than in the control group (376 ± 22, 315 ± 31, t = 3.14, 2.12, respectively, both P < 0.01) ; 3D cell culture assay showed that the cell morphology markedly changed, and the invasive pseudopodia disappeared in the E2F6 inhibition group. Flow cytometry revealed that the proportion of cells at G0-G1 phase and apoptosis rate were significantly higher in the E2F6 inhibition group than in the control group (both P < 0.001). Western blot analysis showed significantly decreased protein expression of β-catenin, activated β-catenin and its downstream target proteins c-Myc and cyclin D1, but significantly increased protein expression of P21 in the E2F6 inhibition group compared with the control group (all P < 0.001) ; additionally, the E2F6 inhibition group showed significantly decreased protein expression of epithelial-mesenchymal transition-related molecules vimentin and N-cadherin, but significantly increased expression of E-cadherin compared with the control group (all P < 0.001) . Conclusions:The E2F6 transcription factor is highly expressed in malignant melanoma. Knockdown of the E2F6 gene in A375 cells can inhibit cell proliferation, migration and invasion by antagonizing the β-catenin signaling pathway.

Diagnostic approach and management of tuberculous meningitis: a nationwide cross-sectional survey in 49 hospitals in China / 中华传染病杂志

Objective:To take a broad overview of the current allocation of diagnosis and treatment resources and management for patients with tuberculous meningitis (TBM) in 49 hospitals in China.Methods:A cross-sectional survey about TBM was carried out in 49 hospitals from 27 provinces across China, by means of electronic questionnaire.The electronic questionnaire was filled by doctors in charge of the departments where TBM patients were routinely admitted from September to December 2018. The availability of medical resources, diagnosis, evaluation, treatment and surveillance in these hospitals were analyzed from the questionnaire. The count data were expressed as percentage.Results:Among the 49 participating hospitals, 37(75.5%) hospitals had less than 50 admissions of suspected TBM per year. Less than 20 TBM patients were confirmed by etiological diagnosis per year in 42(85.7%) participating hospitals.The availability of conventional medical imaging including computed tomography (CT), magnetic resonance imaging (MRI), enhanced MRI, cerebral angiography and magnetic resonance angiography (MRA) were 100.00%(49/49), 95.92%(47/49), 91.84%(45/49), 61.22%(30/49) and 67.35%(33/49), respectively. The rate of access to classic etiological diagnostic methods including acid-fast bacilli smear, mycobacterial culture and T cell spot test of tuberculosis infection were 77.55%(38/49), 95.92%(47/49) and 83.67%(41/49), respectively. Rifampin (100.0%, 49/49), isoniazid (100.0%, 49/49), pyrazinamide (98.0%, 48/49) and ethambutol (95.9%, 47/49) were most commonly used in initial anti-tuberculosis treatment of non-severe patients with TBM. The course of anti-tuberculosis treatment was 18 months in 25(51.0%) hospitals, and 12 months in 17(34.7%) hospitals. Intrathecal glucocorticoid and isoniazid were used in 39(79.6%) hospitals. Dexamethasone was used as part of treatment in 24(49.0%) hospitals, and the duration of glucocorticoid was about two months in 28(57.1%) hospitals. As for hyponatremia, 32(65.3%) hospitals didn′t investigate the cause, and hypertonic saline (83.7%, 41/49) and oral rehydration salts (71.4%, 35/49) were considered as the most common treatment strategy. Lumbar puncture was most commonly used for intracranial pressure surveillance in 48(98.0%) hospitals.Conclusions:The TBM cases admitted to the investigation hospitals are characterized by scattered sources and few confirmed cases of etiology. There are obvious heterogeneities in the diagnosis and treatment of TBM and the management of complications.The standardized plan for diagnosis and treatment of TBM are needed to improve the management.

PMN-CD11b Levels and Its Correlation with Left Atrial Diameter in Patients with Atrial Fibrillation / 中山大学学报(医学科学版)

@#【Objective】To observe the levels of CD11b expressions in the surface of neutrophils in serum，and its correlation with left atrial diameter ，and to study the inflammatory mechanisms in atrial fibrillation（AF） patients. 【Methods】Clinical characteristics and blood samples of AF group and sinal rhythm group were collected. CD11b levels in the surface of neutrophils were examined by flow cytometry. Left atrial diameter was examined by echocardiography. 【Results】The AF group included 85 patients ：36 with paroxysmal AF；26 with persistent AF；23 with permanent AF. The sinal rhythm group includes 57 patients. PMN- CD11b levels were significantly higher in Af group than in sinal rhythm group（P＜0.01）. The left atrial diameter was larger in AF group than in sinal rhythm group（P＜0.01）. Multivariate analysis revealed that PMN-CD11b，left atrial diameter and Valvular heart disease were independent predictors of atrial fibrillation.【Conclusions】PMN- CD11b levels were elevated in paroxysmal AF when AF was present and in persistent， permanent AF patients，implying atrial fibrillation was closely related to inflammation；PMN-CD11b were correlated with left atrial diameter，inflammation might participate in the atrial structural remodeling in AF patients ；PMN-CD11b levels were elevated in AF patients with high risk thrombosis，inflammation might have some value to embolic risk stratification according to CHA2DS2-VASc score.

The application value of afterload-related cardiac performance in patients with sepsis-induced cardiomyopathy / 中华急诊医学杂志

Objective To investigate the application value of afterload-related cardiac performance(ACP) in patients with sepsis-induced cardiomyopathy.Methods A total of 148 patients with septic shock admitted by the department of critical care of Wuhan fourth hospital from April 2013 to March 2018 were retrospectively included,all included patients were divided into LVEF ＜ 50％ group and LVEF ≥ 50％ group according to left ventricular ejection fraction (LVEF),the mortality rate at 28 days and ACP value were compared in the 2 groups.All included patients were divided into normal group (ACP ＞ 80％),mild heart function impairment group (60％ ＜ ACP ≤ 80％),moderate heart function impairment group (40％ ＜ ACP ≤ 60％),and severe heart function impairment group (ACP ≤ 40％) according to ACP value,the mortality rate at 28 days was compared in the 4 groups.The measurement data were compared by grouped t test,the rates were compared by chi-square test.Results The mortality rate was 58.2％ in the LVEF ＜ 50％ group,and 30.9％ in the LVEF ≥ 50％ group,with statistically significant differences (x2=11.171,P＜0.01).The values of ACP in the LVEF ＜ 50％ group were (39.3±16.4) ％,and those in the LVEF ≥ 50％ group were (69.1±14.9) ％,with statistically significant differences (t=l 1.571,P＜0.01).The mortality rate was 14.81％ in the normal group,44.00％ in the mild,58.82％ in the moderate and 90.00％ in the severe group.The differences between the normal group and the mild and moderate groups were statistically significant,while those between the severe group and the mild and moderate groups were statistically significant.Conclusion Aflerload-related cardiac performance is of great value for the diagnosis and prognosis in patients with sepsis-induced cardiomyopathy.

Observation on time-effect for the stimulating at Zusanli (ST 36) with minimally invasive embedding with PGLA in the healthy person / 中国针灸

OBJECTIVE@#To explore the objectivity and time-effect of stimulating effect at acupoint with PGLA in the healthy person, and to provide a basis for the rational interval of minimally invasive embedding of PGLA.@*METHODS@#Before embedding, 8 h, 3rd, 7th, 10th, 14th day after embedding, medical imaging magnetic resonance imaging (MRI) scanning technique was used to collect local T2WI pressure-lowering and T2-Mapping 8 echoes sequence image of left Zusanli (ST 36) in 8 cases of healthy person. The T2-Mapping 8 echoes sequence image was generated by the relevant software to the T2-Mapping image and the local T2 value was measured. The characteristics of local T2WI pressure-fat image signal intensity and the change of T2 value at left Zusanli (ST 36) with minimally invasive embedding with PGLA were observed and analyzed.@*RESULTS@#①There was no abnormal signal on the T2WI pressure-fat image on the left Zusanli (ST 36) point before the embedding. The high-signal was seen on the local T2WI pressure-fat image at each time point after embedding, there was no significant difference in local signal intensity between 8 h, 3rd and 7th day after embedding. The local signal intensity decreased on the 10th day after embedding, and the local signal intensity decreased significantly on the 14th day after embedding.②The T2 value at each time point after embedding increased significantly compared with that before embedding (all 0.05); there was no significant difference between the T2 value on the 7th and the 10th day after embedding (>0.05)，the T2 value on the 14th day after embedding was significantly lower than that on the 7th day after embedding (<0.01).@*CONCLUSION@#It has a stimulating effect on the local acupoints with minimally invasive embedding with PGLA in the healthy person, and the stimulating effect has certain time-effect. The effective stimulation time is about 2 weeks. The rational interval period for the minimally invasive embedding with the PGLA of the same specification type should be about 2 weeks.

The application value of afterload-related cardiac performance in patients with sepsis-induced cardiomyopathy / 中华急诊医学杂志

Objective@#To investigate the application value of afterload-related cardiac performance(ACP) in patients with sepsis-induced cardiomyopathy.@*Methods@#A total of 148 patients with septic shock admitted by the department of critical care of Wuhan fourth hospital from April 2013 to March 2018 were retrospectively included, all included patients were divided into LVEF < 50% group and LVEF≥50% group according to left ventricular ejection fraction (LVEF), the mortality rate at 28 days and ACP value were compared in the 2 groups. All included patients were divided into normal group (ACP > 80%), mild heart function impairment group (60% < ACP≤80%), moderate heart function impairment group (40% < ACP≤60%), and severe heart function impairment group (ACP≤40%) according to ACP value, the mortality rate at 28 days was compared in the 4 groups. The measurement data were compared by grouped t test, the rates were compared by chi-square test.@*Results@#The mortality rate was 58.2% in the LVEF < 50% group, and 30.9% in the LVEF≥50% group, with statistically significant differences (χ2=11.171, P<0.01). The values of ACP in the LVEF < 50% group were (39.3±16.4) %, and those in the LVEF≥50% group were (69.1±14.9) %, with statistically significant differences (t=11.571, P<0.01).The mortality rate was 14.81% in the normal group, 44.00% in the mild, 58.82% in the moderate and 90.00% in the severe group. The differences between the normal group and the mild and moderate groups were statistically significant, while those between the severe group and the mild and moderate groups were statistically significant.@*Conclusion@#Afterload-related cardiac performance is of great value for the diagnosis and prognosis in patients with sepsis-induced cardiomyopathy.

Genetic diversity analysis and germplasm identification of Paris species by CDDP markers / 中草药

Objective: To explore genetic diversity and phylogenic relationship among different types of Paris species and provide an effective method for the rapid identification of germplasm resources. Methods: CDDP marker was used to evaluate the genetic diversity and phylogenic relationship among 13 types of Paris species. And the coding of Paris genus plants was carried out based on method of CODE128 barcode. Results: Our results indicated that 73 polymorphic bands were amplified by 11 primers among 80 bands, and the ratio of polymorphic band was 91.25%. The observed number of alleles (Na), effective number of alleles (Ne), Nei’s gene diversity (H), and Shannon’s information index (I) was 1.912 5, 1.589 6, 0.342 3, and 0.507 0, respectively. The results of UPGMA analysis showed that there were great differences among 13 types of Paris species on genetic diversity. Based on CDDP markers, 13 barcode molecular identity cards were constructed for Paris species based on method of CODE128 barcode. Conclusion: There was abundant polymorphism among Paris species, and CDDP markers were effective to analyze the genetic diversity of Paris species, and the established barcode molecular identity for Paris species was sensitive and fast, which can be used for scientific research and industrial production of Paris genus.

Improving the dissolution rate of water-insoluble diflunisal by γ-cyclodextrin metal-organic framework / 药学学报

The aim of this study is to prepare porous γ-cyclodextrin metal-organic framework (CD-MOF) with good biocompatibility to improve the in vitro release properties of water-insoluble drugs. Different sizes of CD-MOF were obtained by controlling the self-assembly of γ-cyclodextrin and potassium ion and the rate of crystal growth. The poorly water-soluble diflunisal (DIF) was selected as the model drug and loaded into the interior of porous CD-MOF by the impregnation method. The DIF loaded CD-MOF (DIF-MOF) was characterized by scanning electron microscopy (SEM), powder X-ray diffraction (PXRD), nitrogen adsorption and desorption, Fourier infrared spectrometer and thermogravimetric analysis. In addition, in vitro cytotoxicity and solubilizing capability of CD-MOF were investigated. It revealed that the obtained CD-MOF was cubic-like with a narrow size distribution and high porosity. Negligible cytotoxicity was found after incubation with RAW264.7 cells. Compared with the pure CD-MOF carrier, the morphology and crystal form of DIF-MOF was not damaged during the drug loading process. Moreover, the solubility and release rate of water-insoluble DIF from the DIF-MOF were significantly increased.

MGST1 overexpression promotes hepatocellular carcinoma development / 基础医学与临床

Objective To investigate the expression of microsomal glutathione S-transferase 1 ( MGST1) in hepa-tocellular carcinoma ( HCC) and its significance in the development of HCC. Methods Western blot was used to measure MGST1 expression in human hepatocellular carcinoma and adjacent tissues and HCC cell lines. Further-more, shRNA targeting MGST1 was constructed and transected into MHCC97H and HCCLM3 cells to deplete MGST1 expression. MGST1 was over-expressed in SK-Hep-1 cells using pCDH lentivirus system. Cell proliferation and migration were analyzed by colony formation and Transwell migration assay, respectively. The subcutaneous xenograft model of MHCC97H cells in nude mice was established to check tumor development and mouse survival.Results MGST1 was higher in 71% (17/24) of HCC tissues compared with their adjacent liver tissues. Cell proliferation and migration were significantly decreased by MGST1 knockdown, while they were increased by MGST1 overexpression. Furthermore, mice implanted with shMGST1 MHCC97H cells exhibited retarded tumor formation and tumor progression compared with control group. Conclusions MGST1 overexpression promotes hepatocellular carcinoma development and this molecule targeted for HCC treatment.