ObjectiveTo investigate the role and mechanism of hyodeoxycholic acid （HDCA） in the progression of metabolic associated fatty liver disease （MAFLD）， and to provide a new theoretical basis for further clarifying the pathogenesis of MAFLD. MethodsL02 hepatocytes were used as experimental cells， and palmitic acid was used to induce steatosis in L02 cells. The farnesoid X receptor （FXR） siRNA interference chain technique was used to construct a hepatocyte cell line with low FXR expression. CCK8 assay was used to observe the effect of HDCA on L02 steatosis hepatocytes at different concentrations （0， 100， 200， 300， and 400 μmol/L） and time points （12， 24， 36， and 48 hours）. The method of qRT-PCR was used to measure the mRNA expression levels of FXR， proliferating cell nuclear antigen （PCNA）， Cyclin D1， phosphatidylinositol 3-kinase （PI3K）， and protein kinase-B （AKT）， and Western blot was used to measure the protein expression levels of FXR， Cyclin D1， PCNA， PI3K， phosphorylated PI3K （p-PI3K）， AKT， and phosphorylated （p-AKT）. A one-way analysis of variance was used for comparison of normally distributed continuous data with homogeneity of variance between multiple groups， and the Tukey HSD test was used for further comparison between two groups； the Welch analysis of variance was used for comparison of normally distributed continuous data with heterogeneity of variance between multiple groups， and the Games-Howell test was used for further comparison between two groups. The independent-samples t test was used for comparison between two groups. ResultsCCK8 assay showed a significant reduction in the viability of L02 cells and steatosis hepatocytes treated by 300 μmol/L HDCA （P<0.05）， and qRT-PCR showed a significant increase in the mRNA expression level of FXR and significant reductions in the mRNA expression levels of PCNA， Cyclin D1， PI3K， and AKT （all P<0.05）. Western blot showed a significant increase in the protein expression level of FRX （P<0.05）， and after interference of FXR expression in L02 cells， there were significant increases in the protein expression levels of PCNA， PI3K， p-PI3K， AKT， and p-AKT （all P<0.05）. ConclusionHDCA inhibits the PI3K/AKT signaling pathway by upregulating FXR expression， thereby inducing a reduction in the viability of steatosis hepatocytes.

To preliminarily understand the pathogenic mechanism of Mycoplasma genitalium (Mg) GroEL protein, we used bioinformatics tools to predict the structure and function of Mg GroEL protein and then constructed the recombinant plasmid pET-28a-GroEL. The protein expression was induced by 0.2 mmol/L IPTG, and the expressed protein was purified by Ni-iminodicitic acid (IDA) column affinity. Tohoku Hospital Pediatrics-1 (THP-1) cells were exposed to 2 μg/mL Mg rGroEL. The levels of interleukin (IL)-1β and tumor necrosis factor (TNF)-α in the cell supernatant were measured by ELISA, and that of IL-6 was measured by an automatic chemiluminescence instrument. The activation of the nuclear factor-kappa B (NF-κB) signaling pathway was visualized by immunofluorescence and Western blotting. The results showed that Mg GroEL was a stable hydrophilic protein composed of 543 amino acid residues, with the relative molecular mass of 58.44 kDa, an isoelectric point of 5.68, and a molecular formula of C₂₅₆₈H₄₃₀₀N₇₀₀O₈₂₅S₈. The secondary structure was mainly composed of α-helices and random coils. Mg GroEL contained 12 B-cell dominant epitopes and 10 T-cell dominant epitopes. It exhibited high homology with the GroEL proteins from Mycoplasma pneumoniae, M. agalactiae, M. arthritidis, M. hyopneumoniae, and M. bovis. Mg rGroEL activated the NF-κB signaling pathway and promoted the secretion of IL-1β, IL-6, and TNF-α in THP-1 cells. These results suggest that Mg GroEL exhibits substantial antigenicity and possesses the capability of triggering inflammation in host cells. This study establishes a theoretical basis for future investigations pertaining to the role and pathogenic mechanisms of Mg GroEL.
Mycoplasma genitalium/metabolism* ; Chaperonin 60/metabolism* ; Computational Biology ; Bacterial Proteins/genetics* ; Humans ; NF-kappa B/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Interleukin-1beta/genetics* ; Inflammation ; Interleukin-6/genetics* ; Recombinant Proteins/genetics* ; THP-1 Cells ; Signal Transduction ; Escherichia coli/metabolism*

Metabolic associated fatty liver disease (MAFLD) has become a prevalent chronic liver disease worldwide because of lifestyle and dietary changes. Gut microbiota and its metabolites have been shown to play a critical role in the pathogenesis of MAFLD. Understanding of the function of gut microbiota and its metabolites in MAFLD may help to elucidate pathological mechanisms, identify diagnostic markers, and develop drugs or probiotics for the treatment of MAFLD. Here we review the pathogenesis of MAFLD by gut microbiota and its metabolites and discuss the feasibility of treating MAFLD from the perspective of gut microbes.
Gastrointestinal Microbiome ; Fatty Liver/microbiology* ; Humans

Objective To investigate the role of complement in radiation-induced lung injury in mice after chest irradiation with ⁶⁰Co γ-rays at a single dose of 20 Gy. Methods C57BL/6 mice underwent chest irradiation with ⁶⁰Co γ-rays at a single dose of 20 Gy, followed by observation for the inflammatory reaction of the lung tissue in the early stage (within 15 d) and pulmonary fibrosis in the later stage (30 and 180 d). Enzyme-linked immunosorbent assay was used to measure the levels of C2, C3a, C4, and C5b-9 in the lung tissues at 1, 3, 7, 15, 30, and 180 d after irradiation. The expression of complement mRNA in BEAS-2B cells after irradiation was determined using RT-PCR. Results Radiation-induced lung injury in micepresented as inflammatory response in the early stage and fibrosis in the late stage. Complement C2, C4, and C5b-9 complexes were increased in the early period (3 or 7 d) after irradiation (P < 0.05), which might be associated with the inflammatory response induced by irradiation. During 3 to 180 d, complement C3a was significantly higher in the irradiated mice than in the control mice, suggesting a close relationship between C3a and radiation-induced lung injury. The irradiated cells showed increased mRNA expression of C2 and C3, with no changes in the mRNA levels of C4 and C5. Conclusion Different complement proteins have varying responses to radiation-induced lung injury, among which C3a is closely related to radiation-induced lung injury, suggesting that regulating C3a and its receptors may be a new way to prevent and treat radiation-induced lung injury.

BACKGROUND: Prefabricated composite tissue flaps have been used by Bakamjjan for cardiac repair since 1973, but have not been widely used due to technique limitations. Domestic research on prefabricated composite tissue flaps to repair limb bones is rarely reported. OBJECTIVE: To analyze the effects of prefabricated composite tissue flaps in the repair of limb bone and corresponding soft tissue defects. METHODS: New Zealand big rabbit models of bone fracture and soft tissue defect of the extremities were constructed (first operation) and randomly divided into three groups. In group A, prefabricated composite tissue flap was used to treat bone and soft tissue defects of the extremities at 10 days after modeling (second operation). In group B, free femur was used to treat bone and soft tissue defects of the extremities at 10 days after modeling. In group C, the incision was open and sutured with no treatment. General condition, body weight, imaging finding and histological findings were compared between groups. RESULTS AND CONCLUSION: There was 100% survival in all the three groups. Graft displacement was observed in 2 rabbits in the group B, but the deformity healed, which had little effect on the rabbit's mobility. The weight recovery and gain were higher in the group A than in the group B (P < 0.05). The imaging findings showed that a large number of calluses were formed in the group A at 2 weeks after operation, which were bridged in gap at 4 weeks after operation, filled in the defect gap at 8 weeks, and remodeled at 12 weeks. In the group B, a small amount of calluses were formed at 2 weeks after operation, and began to increase at 4 weeks. The femoral cut was obvious. A large number of calluses were formed at 8 weeks after operation, and the defect gap was filled at 12 weeks after operation. In the group C, the callus began to form at 8 weeks after operation, and the defect gap was still present, with osteosclerosis at the two ends. The Lane-Sandhu score was statistically different between the three groups at 8 and 12 weeks after second operation (P < 0.05). Histological observation indicated that a large number of newly formed osteoblasts and bone cells were formed in the group A at 4 weeks after operation, and the tubular structure increased and irregular bone island formed at 8 weeks; new bone formed at 12 weeks, with the presence of the medullary cavity containing yellow bone marrow dominated by adipocytes. In the group B, most of the grafted bones were degraded and absorbed at 4 weeks after operation, and osteoblasts were ossified at 8 weeks. The remaining implanted bones were still visible. Most of the osteoblasts were ossified and became lamellar bones at 12 weeks. In the group C, the defect area was filled with a large amount of fibrous connective tissues at 12 weeks after operation, and there was no bone formation. To conclude, the prefabricated composite tissue flap can be used to repair the bone and soft tissue defects of the extremities, and it has a faster and better therapeutic effect than the traditional free bone repair.

Chloroquine is a quinine derivative which is synthesized by German scholars in 1934. In addition to its anti-malaria, treatment of systemic lupus erythematosus and immunomodulatory effects, chloroquine is also found valuable in broad-spectrum antiviral treatment. Clinical trials have confirmed that chloroquine has a good effect on acquired immunodeficiency syndrome. In 2019, there were many patients infected with novel coronavirus (severe acute respiratory syndrome coronavirus 2, SARS-CoV-2). Preliminary clinical trials showed that chloroquine had obvious curative effect on patients with SARS-CoV-2. We summarize the effects of chloroquine to different viruses, explain its mechanism, and compare the results of its experiments in vitro and in vivo. The antiviral effect of chloroquine in vivo and in vitro are not consistent, which may be related to the model of animal, dosage and distribution of chloroquine in vivo, and the design of clinical research.

ObjectiveTo study the mechanism of renal injury in urogenic sepsis and explore the effect of H2S on the expression of p38 mitogen-activated protein kinase (p38MAPK) and Cysteine protease 3(Caspase3) and apoptosis in renal tissue of urogenic sepsis. Hydrogen sulfide (H2S) has a wide range of biological effects and has a certain protective effect on the kidney. The main objective of this study is to investigate the effect of H2S on the expression of p38 mitogen-activated protein kinase (p38MAPK), cysteine proteinase 3 (Caspase3), and cell apoptosis in renal tissue of urogenic sepsis, and further to study the mechanism of urinary sepsis renal injury.MethodsNew Zealand rabbits (n=40) were divided into five groups Control, Sham, Sepsis, NaHS, and PAG, with eight rabbits in each group. The vital signs, blood routine white blood cell count, neutrophil count, and renal function (Cr, BUN) of five groups of New Zealand rabbits were recorded before the operation, 24 hrs after the operation, 48 hrs after the operation, and 72 hrs after the operation. 72 hrs after the operation, the left kidney tissue was taken for HE staining to observe the changes of cell morphology and structure of the kidney tissue. The apoptotic cells in renal tissue were labeled by Tunel assay (in situ terminal transferase labeling technique). The expression levels of p38MAPK and Caspase3 in renal tissue were tested by ELISA.Results The apoptosis indexes of renal tissue cells in group Control and group Sham were (5.65±2.43)% and (5.57±2.72)%, respectively, with no statistically significant difference (P>0.05). The apoptosis index of rabbit kidney tissue in group Sepsis was (25.26±2.70)%, which was significantly higher than that in group Control and group Sham (P<0.05). The apoptosis index (16.93±2.03)% in group NaHS was significantly lower than that in group Sepsis (P<0.05). The apoptosis index of group PAG (33.09±3.70)% was significantly higher than that of group Sepsis and group NaHS (P<0.05). There was no significant difference between group Control and group Sham (P>0.05). Pairwise comparison between groups Sepsis, NaHS, and PAG showed statistically significant differences (P<0.05). The expression levels of group Sepsis and group PAG were significantly higher than that of group NaHS (P<0.05). The expression level of group PAG was significantly higher than that of group Sepsis (P<0.05).Conclusion H2S can alleviate renal damage caused by urine-derived sepsis, and its mechanism combined with H2S to suppress the expression of p38MAPK and Caspase3 in the renal tissue of urogenous sepsis, thereby reducing renal cell apoptosis Death.

Objective: To construct 3β-HSD gene shRNA lentivirus interference vecto, then transfect into human MCF-7 cells, and construct cell line with 3β-HSD gene silencing, finally to study the effects of 3β-HSD on apoptosis induced by di- (2-ethylhexyl) phthalate (DEHP) . Methods: According to the mRNA sequence of 3β-HSD gene provided by GenBank, three interference sequences were designed and connected to PLVX-shRNA2-puro after annealing. The recombinant lentivirus vector was transfected into 293FT cells, the virus supernatants were collected and infected with MCF-7 cells. After puromycin screening, MCF-7 cells with 3β-HSD gene silencing were constructed. The cells with 3β-HSD gene silencing were identified by real-time quantitative PCR and western blot. Then the 3β-HSD gene silencing cells and MCF-7 cells were treated at various doses of DEHP for 24 hours to detect the gene expression and protein expression of apoptosis genes including Bax, Caspase-3 and Caspase-8. Results: The interference sequence of 3β-HSD gene inserted into lentivirus vector PLVX-shRNA2-puro is consistent with the designed sequence. 3β-HSD gene expression level in MCF-7 cells with 3β-HSD gene silencing was 77% lower than than that of control MCF-7 cells. 3β-HSD protein level in MCF-7 cells with 3β-HSD gene silencing was 74% lower than that of control MCF-7 cells. After DEHP treatment in MCF-7 cells with 3β-HSD gene silencing and control MCF-7 cells, qRT-PCR results showed that Bax gene expression levels increased by 28%-54%, Caspase-3 gene increased by 13%-49%, Caspase-8 gene increased by 21%-70% in MCF-7 cells when compared with the control group. Additionally, in the 3β-HSD gene silencing cells, Bax gene expression level decreased by 11%-28%, Caspase-3 gene expression decreased by 12%-23%, Caspase-8 gene expression decreased by 11%-34%, compared with the same treatment group of MCF-7 cells. Western blot results showed that Bax protein expression level increased by 28%-61%, Caspase-3 protein expression level increased by 40%-48%, Caspase-8 protein increased by 31%-84% in MCF-7 cells when compared with the control group. In 3β-HSD gene silencing cells, Bax protein expression level increased by 11%-27%, Caspase-3 protein increased by 21%-40%, Caspase-8 protein increased by 12%-25%, compared with the same treatment group of MCF-7 cells. Conclusion The stable 3β-HSD gene silencing cell line are successfully constructed in this study. DEHP can induce increased expression of apoptotic gene and protein. Silencing of 3β-HSD gene can inhibit the activation of apoptotic gene by DEHP in a certain degree.

Objective To analyze the differences in social psychological characteristics of patients with different subtypes of functional dyspepsia (FD).Methods From August 2011 to July 2015,210 FD patients met Rome Ⅲ criteria were enrolled and divided into pure postprandial distress syndrome (PDS)group (69 cases),pure epigastric pain syndrome (EPS) group (74 cases) and PDS overlap EPS (PDS+ EPS) group (67 cases).Hamilton depression scale (HAMD),Hamilton anxiety scale (HAMA),life event scale (LES) and Eysenck's personality questionnaire (EPQ) were used for evaluation.Chi-square test and least-significant difference were performed for statistical analysis.Results Incidence rates of depression and anxiety of PDS+EPS group were both 100.0％ (67/67),which were higher than those of pure PDS group (84.1％,58/69 and 91.3％,63/69),and the differences were statistically significant (x2 =11.62 and 16.34,both P＜0.01);which were also higher than those of pure EPS group (78.4％,58/74 and 90.5％,67/74),and the differences were statistically significant (x2 =6.10 and 6.67,both P＜0.05).Somatic anxiety score and mental anxiety score of PDS+ EPS group were 16.34±3.70 and 14.18±2.99,respectively;which were higher than those of pure PDS group (11.26±3.42 and 10.70±2.94) and pure EPS group (12.30 ± 4.29 and 10.36 ± 2.63),and the differences were statistically significant (t=8.33,5.97,6.85 and 8.06;all P＜0.01).The top two life events in three groups were sleeping habits alteration (67.6％,142/210) and severe disease or trauma (26.7％,56/210).The incidence rate of sleeping habits alteration in pure PDS group was 53.6 ％ (37/69),which was lower than that in pure EPSgroup (77.0％,57/74) and PDS+EPS group (71.6％,48/67),and the differences were statistically significant (x2 =8.68 and 4.71,both P＜0.05).Most of P scale scores of pure PDS group,pure EPS group and PDS+EPS group were normal,most of E and L scale scores were low;most of N scale scores of pure PDS group were low,and these of pure EPS group and PDS+EPS group were normal.Conclusions The incidence of depression and anxiety of PDS+EPS group is the highest.Somatic anxiety is more obvious than mental anxiety in PDS+EPS group and pure EPS group,most with sleep events.Slow emotional response is common in pure PDS group.

Objective To observe the clinical efficacy of acupuncture plus Chinese medicinal fumigation in treating chronic pelvic inflammatory disease (CPID).Method Totally 120 CPID patients were randomized into group A, group B and group C, 40 cases in each group. Group A was intervened by acupuncture plus Chinese medicinal fumigation, group B was by dry acupuncture treatment, while group C was by Chinese medicinal fumigation alone. After 3 treatment courses, the clinical efficacies were observed, and the relapse rates among the cured cases in 8 months after the whole treatment were compared among the 3 groups.Result The total effective rate and recovery rate were respectively 95.0% and 70.0% in group A, versus 82.5% and 45.0% in group B, and 57.5% and 32.5% in group C, and the total effective rate and recovery rate in group A were significantly different from that in group B and group C (P<0.05). The treatment duration for the recovered cases in group A was significantly different from that in group B and C (P<0.05). The relapse rate in the recovered cases in the 8-month follow-up was 10.7% in group A, versus 44.4% in group B and 53.8% in group C, and the relapse rate in group A was markedly lower than that in group B and C (P<0.05).Conclusion Acupuncture plus Chinese medicinal fumigation is an effective method in treating CPID, and its advantages include content efficacy, short treatment duration, and low relapse rate, etc.