Objective To investigate the therapeutic effect of Liangxue-jiedu decoction on acute-on-chronic liver failure(ACLF)model mice and the Toll-like receptor 4(TLR4)/c-Jun amino terminal kinase(JNK)/nuclear factor κB(NF-κB)signaling pathway.Methods An ACLF mouse model was established using combined treatment of carbon tetrachloride,lipopolysaccharide,and D-galactosamine.Biochemical analysis was performed to evaluate liver function indicators,including alanine aminotransferase,aspartate aminotransferase,and total bilirubin.Histopathological examination was conducted to assess liver tissue morphological changes.Quantitative PCR was used to detect the mRNA expression of tumor necrosis factor alpha(TNF-α),interleukin(IL)-6,IL-1 β,and TLR4 in liver tissues.A CCK8 assay was used to evaluate the optimal interventional concentration of Liangxue-jiedu decoction on Raw 264.7 cells.Enzyme-linked immunosorbent assay was used to detect the contents of TNF-α,IL-6,and IL-1βin the cell supernatant.Protein immunoblotting was performed to measure the expression of TLR4/JNK/NF-κB signaling pathway-related proteins,including TLR4,NF-κB,p-ERKl/2,ERK1/2,p-JNK,and JNK.Results Compared with the ACLF model group,the Liangxue-jiedu decoction-treated group showed reduced cell necrosis,fibrosis,and inflammatory cell infiltration in liver tissues;decreased serum levels of alanine aminotransferase,aspartate aminotransferase and total bilirubin;and lower expression of TNF-α,IL-6,IL-1β,and TLR4 mRNA.Liangxue-jiedu decoction reduced TLR4/JNK/NF-κB signaling pathway-related protein expression in liver tissues.The in vitro result also showed that Liangxue-jiedu decoction reduced TNF-α,IL-6,and IL-1 β secretion by macrophage cells and down-regulated the expression of TLR4/JNK/NF-κB signaling pathway proteins.Conclusions Liangxue-jiedu decoction effectively improved liver function in ACLF mice in a manner closely related to the downregulation of TLR4/JNK/NF-κB pathway proteins.

OBJECTIVE: To explore the genetic etiology for a child with profound intellectual disabilities and obvious behavioral abnormalities. METHODS: A male child who had presented at the Zhongnan Hospital of Wuhan University on December 2, 2020 was selected as the study subject. Peripheral blood samples of the child and his parents were collected and subjected to whole exome sequencing (WES). Candidate variant was verified by Sanger sequencing. Short tandem repeat (STR) analysis was carried out to determine its parental origin. The splicing variant was also validated in vitro with a minigene assay. RESULTS: WES results revealed that the child had harbored a novel splicing variant of c.176-2A>G in the PAK3 gene, which was inherited from his mother. The results of minigene assay have confirmed aberrant splicing of exon 2. According to the guidelines from the American College of Medical Genetics and Genomics, it was classified as a pathogenic variant (PVS1+PM2_Supporting+PP3). CONCLUSION The novel splicing variant c.176-2A>G of the PAK3 gene probably underlay the disorder in this child. Above finding has expanded the variation spectrum of the PAK3 gene and provided a basis for genetic counseling and prenatal diagnosis for this family.
Child ; Female ; Humans ; Male ; Pregnancy ; Exons ; Intellectual Disability/genetics* ; Mothers ; Mutation ; p21-Activated Kinases/genetics* ; Parents ; RNA Splicing

PURPOSE: Numerous studies have assessed the association of SP110 gene variants with tuberculosis (TB), but the results were inconsistent. Through a comprehensive review and meta-analysis, our study aimed to clarify the nature of genetic risks contributed by 11 polymorphisms for the development of TB. MATERIALS AND METHODS: Through searching PubMed, web of science, China National Knowledge Infrastructure (CNKI) databases, a total of 11 articles including 13 independent studies were selected. The pooled odd ratios (ORs) along with their corresponding 95% confidence interval (CI) were estimated for allelic comparisons, additive model (homozygote comparisons; heterozygote comparisons), dominant model and recessive model. We also assessed the heterogeneity across the studies and publication bias. RESULTS: The results of combined analysis revealed a significantly increased risk of TB for single nucleotide polymorphism (SNP) rs9061 in all five comparisons (allelic comparisons: OR=1.28, 95% CI=1.14–1.44, p<0.0001; homozygote comparisons: OR=2.84, 95% CI=1.84–4.38, p<0.00001; heterozygote comparisons: OR=1.23, 95% CI=1.05–1.43, p=0.009; dominant model: OR=1.32, 95% CI=1.14–1.53, p=0.0003; recessive model: OR=2.26, 95% CI=1.18–4.34, p=0.01). In subgroup analysis, the risk of TB associated with SNP rs9061 appeared to be increased. Moreover, increased risk of TB was also found in Asian subgroup of SNP rs11556887, while decreased risk of TB appeared in large sample size subgroup of SNP rs1135791. No significant association was observed between other SNPs and the risk of TB. CONCLUSION: Our meta-analysis suggested that the variant of SNP rs9061 might be a risk factor for TB.
Alleles ; Asian Continental Ancestry Group/genetics ; China ; Confidence Intervals ; Genetic Predisposition to Disease ; Heterozygote ; Homozygote ; Humans ; Minor Histocompatibility Antigens/*genetics ; Nuclear Proteins/*genetics ; Odds Ratio ; *Polymorphism, Single Nucleotide ; Risk Factors ; Tuberculosis, Pulmonary/*genetics

OBJECTIVETo analyze relationships between the tumor deposits (TD) and clinicopathologic features of gastric cancer and investigate the value of TD in staging and prognosis in gastric cancer patients.

METHODSRetrospective cohort study was conducted to evaluate the clinicopathologic data of 388 gastric cancer patients who underwent surgical procedures in Chinese PLA General Hospital between November 2011 and December 2012. Relationships between TD and clinicopathologic features were analyzed by χor Fisher exact tests. Survival curves were also generated by Kaplan-Meier method. The univariate and multivariate analysis were performed with Log-rank and COX proportional hazard model to examine the association between prognosis and TD.

RESULTSTD were observed in 67 (17.3%) of 388 gastric cancer patients, including 48 male patients (48/289, 16.6%) and 19 female patients (19/99, 19.2%). There were 40 patients (40/198, 20.2%) whose age was above 64 years old. TNM staging of positive TD patients was as follows: for pathology, there were 5 patients (5/64, 7.8%) in stage II(b, 6 patients (6/58, 10.3%) in stage III(a, 14 patients (14/75, 18.7%) in stage III(b, 30 patients (30/135, 22.2%) in stage III(c, 12 patients (12/39, 30.8%) in stage IIII( and no one in stage I(b or II(a; for T-staging, there were 2 patients (2/18, 11.1%) in stage T2, 2 patients (2/27, 7.4%) in stage T3, 36 patients (36/259, 13.9%) in stage T4a and 27 patients (27/84, 32.1%) in stage T4b; for N-stage, there were 5 patients (5/72, 6.9%) in stage N0, 6 patients (6/72, 8.3%) in stage N1, 19 patients (19/82, 23.2%) in stage N2, 27 patients (27/100, 27.0%) in stage N3a and 10 patients(10/62, 16.1%) in stage N3b; for M-stage, there were 12 patients (12/40, 30.0%) in distal metastases; for vascular invasion, there were 29 patients (29/129, 22.5%). Among positive TD patients, the number of TD >3 was found in 38 of 67 cases(56.7%). TD was associated with pTNM-stage (χ=16.898, P=0.010), T-stage (χ=17.382, P=0.001), N-stage (χ=18.080, P=0.001), M-stage (χ=5.060, P=0.036) and vascular invasion(χ=3.675, P=0.039). The median survival time of positive TD patients was significantly shorter as compared to negative TD patients (22 months vs. 32 months, χ=23.391, P=0.012). Among positive TD patients, the median survival time of patients with TD number >3 was significantly shorter as compared to those with TD number <3 (17 months vs. 25 months, χ=5.157, P=0.023). Multivariate survival analysis showed that TD number >3 was the independent risk factor of prognosis (RR=2.350, 95%CI:1.345 to 4.106, P=0.003).

CONCLUSIONSTD state is closely associated with the staging of gastric cancer and TD number >3 indicates a poor prognosis.

Aged ; China ; Cohort Studies ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Multivariate Analysis ; Neoplasm Invasiveness ; pathology ; Neoplasm Metastasis ; Neoplasm Staging ; methods ; statistics & numerical data ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Risk Factors ; Stomach Neoplasms ; classification ; diagnosis ; mortality ; pathology ; Survival Rate

OBJECTIVETo study the effect of adenosine preconditioning on cell apoptosis and expressions of glucose-regulated protein (GRP-78) and cysteinyl aspartate-specific protease 12 (caspase-12) in rats with spinal cord ischemia-reperfusion injury.

METHODSTwenty-seven rats were randomized into 3 equal groups and subjected to sham operation (group A), spinal cord ischemia-reperfusion injury (group B), or ischemia-reperfusion injury with adenosine treatment. Spinal cord ischemia-reperfusion injury was induced by cross-clamping of the abdominal aorta inferior to the left renal artery. The spinal cord function was assessed using the Modified Tarlov Scale at 6, 12, and 24 h after reperfusion. At 24 h after reperfusion, histological analysis was carried out with HE staining; cell apoptosis and viability were determined with TUNEL staining, and the expressions of GRP-78 and caspase-12 proteins were determined with Western blotting.

RESULTSHE staining of the spinal cord showed extensive spinal cord injury such as cell edema in group B as compared with group C. Compared with group A, group B showed a significantly increased number of apoptotic cells; the number of apoptotic cells in group B was greater than that in group C. Compared with group B, group C showed significantly increased GRP-78 expression (P<0.01) and decreased caspase-12 expression (P<0.01).

CONCLUSIONAdenosine can up-regulate GRP-78 expression and down-regulate caspase-12 expression, and protects the spinal cord against ischemia-reperfusion injury by inhibiting cell apoptosis.

Adenosine ; pharmacology ; Animals ; Apoptosis ; drug effects ; Caspase 12 ; metabolism ; Heat-Shock Proteins ; metabolism ; Ischemic Preconditioning ; methods ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; Spinal Cord Ischemia ; metabolism

Objective To study the effect of adenosine preconditioning on cell apoptosis and expressions of glucose-regulated protein (GRP-78) and cysteinylaspartate-specific protease 12 (caspase-12) in rats with spinal cord ischemia-reperfusion injury. Methods Twenty-seven rats were randomized into 3 equal groups and subjected to sham operation (group A), spinal cord ischemia-reperfusion injury (group B), or ischemia-reperfusion injury with adenosine treatment. Spinal cord ischemia-reperfusion injury was induced by cross-clamping of the abdominal aorta inferior to the left renal artery. The spinal cord function was assessed using the Modified Tarlov Scale at 6, 12, and 24 h after reperfusion. At 24 h after reperfusion, histological analysis was carried out with HE staining; cell apoptosis and viability were determined with TUNEL staining, and the expressions of GRP-78 and caspase-12 proteins were determined with Western blotting. Results HE staining of the spinal cord showed extensive spinal cord injury such as cell edema in group B as compared with group C. Compared with group A, group B showed a significantly increased number of apoptotic cells;the number of apoptotic cells in group B was greater than that in group C. Compared with group B, group C showed significantly increased GRP-78 expression (P<0.01) and decreased caspase-12 expression (P<0.01). Conclusion Adenosine can up-regulate GRP-78 expression and down-regulate caspase-12 expression, and protects the spinal cord against ischemia-reperfusion injury by inhibiting cell apoptosis.

Objective To study the effect of adenosine preconditioning on cell apoptosis and expressions of glucose-regulated protein (GRP-78) and cysteinylaspartate-specific protease 12 (caspase-12) in rats with spinal cord ischemia-reperfusion injury. Methods Twenty-seven rats were randomized into 3 equal groups and subjected to sham operation (group A), spinal cord ischemia-reperfusion injury (group B), or ischemia-reperfusion injury with adenosine treatment. Spinal cord ischemia-reperfusion injury was induced by cross-clamping of the abdominal aorta inferior to the left renal artery. The spinal cord function was assessed using the Modified Tarlov Scale at 6, 12, and 24 h after reperfusion. At 24 h after reperfusion, histological analysis was carried out with HE staining; cell apoptosis and viability were determined with TUNEL staining, and the expressions of GRP-78 and caspase-12 proteins were determined with Western blotting. Results HE staining of the spinal cord showed extensive spinal cord injury such as cell edema in group B as compared with group C. Compared with group A, group B showed a significantly increased number of apoptotic cells;the number of apoptotic cells in group B was greater than that in group C. Compared with group B, group C showed significantly increased GRP-78 expression (P<0.01) and decreased caspase-12 expression (P<0.01). Conclusion Adenosine can up-regulate GRP-78 expression and down-regulate caspase-12 expression, and protects the spinal cord against ischemia-reperfusion injury by inhibiting cell apoptosis.

Objective: To investigate the role of T cell in the antitumor immune responses induced by MIF gene-modified tumor vaccine. Methods: MIF gene was transferred into FBL3 erythroleukemia cel l by adenovirus carrier and a new type of tumor vaccine was prepared. The chang es of the number and the function of T cell in spleen and lymph node was observe d. Results: After the mice were immunized with MIF gene-m odified FBL3 vaccine, the number of lymphocyte in spleens and lymph nodes increa sed markedly and the specific CTL activities of splenocytes also increased great ly. FACS analysis showed that the CD3+,　CD4+,　CD8+ T cells and CD28 posi tive cells in draining lymph nodes of MIF-FBL3 group mice increased more marked ly than that of control groups. When the wild type FBL3 cells were injected into the mice immunized with MIF gene-modified FBL3 vaccine, the growth of tumors w ere obviously inhibited and the survival rate of the mice was increased. Conclusion: It is suggested that MIF gene-modified tumor vaccine can induce specific antitumor immune responses mediated by T cells and may be a candidate for gene therapy of tumor.

Objective:In order to explore the anti inflammatory mechanisms of ? melanocyte stimulating hormone (? MSH), the effects of ? MSH on the production of NO and proinflammatory cytokines in astrocytes induced by LPS were investigated Methods:Rat brain astrocytes cultured in vitro were stimulated with LPS or given ? MSH with LPS stimulation NO produced in astrocytes was tested with Griess reagent IL 1, IL 6 and TNF ? secreted from astrocytes were examined by MTT assay The expression of macrophage migration inhibitory factor (MIF) mRNA was examined with semiquantitative RT PCR analysis Results:The production of NO, IL 1, IL 6, TNF ? and the expression of MIF mRNA were significantly increased in astrocytes stimulated with LPS If giving ? MSH with LPS stimulation, the production of NO, IL 1, IL 6, TNF ? and the expression of MIF mRNA were markedly decreased Conclusion:[WT5”,6BZ]It is suggested that the inhibitory actions of ? MSH on the production of NO and proinflammatory cytokines in astrocytes are related to the inhibitory effects of ? MSH on inflammation in central nervous system

The in vivo tumorigenicity of murine B16 melanoma cells engineered to secret TNF-a was observed. The retrovirus containing mouse TNF-a cDNA was generated by the virus-packing cell PA317 transfected with plasmid pXT-TNF. The B16 cell clone secreting the highest TNF-a level was obtained after G418 resistance selection, limiting dilution and the assay of TNF-a activity. After the mice were inoculated subcutaneously with the cell clone, we found the tumor growth was inhibited and the survival period of the mice extended when compared with the mice inoculated with the wild-type B16 cells . We also found that the tuinorigenicity of B16-TNF-a+ cell was associated with the cell number inoculated. At or above the 1.25? 104 cells, the percentage of the mice with detectable tumor correlated negatively with the cell number inoculated: however, at the 6.25 ? 103 cells, the percentage was higher than that at 2.5?10~(4) cells. These results encourage us to do further experiments on the following tumor cell-targeted TNF-a gene therapy.