AIM:To investigate the protective effect of hyperoside (Hyp) on Mycoplasma pneumoniae (MP) pneumonia (MPP) mice and the underlying regulatory mechanism.METHODS:BALB/c mice (n=40) were randomly divided into control group, model group, low-dose (12.5 mg/kg) Hyp group and high-dose (50 mg/kg) Hyp group.The MPP model was established using MP standard strains (MPFH) through nasal dripping.The treatment started on the second day, and the follow-up experiments were performed after 3 d.The lung histopathology were detected by HE staining.The levels of C-reactive protein (CRP), alanine aminotransferase (ALT), aspartate transaminase (AST), lactate dehydrogenase (LDH) and MB isoenzyme of creatine kinase (CK-MB) in the serum, and the levels of reactive oxygen species (ROS), superoxide dismutase (SOD) and glutathione (GSH) in the lung tissues were measured by automatic biochemical analyzer.ELISA and Western blot were used to detect interleukine-1 beta (IL-1β), IL-6, IL-18 and tumor necrosis factor-alpha (TNF-α) levels in the serum and lung tissues, respectively.The expression of NLRP3, ASC and caspase-1 at mRNA and protein levels was evaluated by RT-qPCR and Western blot.RESULTS:Compared with control group, the inflammation and alveolar interstitial widening were observed in the lung tissues of model mice, and the CRP, ALT, AST, LDH and CK-MB levels were significantly up-regulated (P<0.05).The ROS level was increased, while SOD and GSH levels were significantly decreased (P<0.05).The levels of IL-1β, IL-6, IL-18 and TNF-α were markedly up-regulated (P<0.05).The protein expression of NLRP3, ASC and caspase-1 was also obviously increased (P<0.05) in model group.Compared with model group, the symptoms above were significantly relieved in Hyp group, and the effect of high-dose Hyp was more apparent than that of the low-dose one.CONCLUSION:Hyperoside suppresses the injury in MPP mice, which may be related to the inhibition of oxidative stress and NLRP3 inflammasome activation.

Objective: To compare the efifcacy and safety of circumferential pulmonary vein antecourt isolation (CPVAI) ablation and stepwise linear (SL) ablation in treating the patients with atrial ifbrillation (AF) Methods: A total of 136 AF patients with catheter ablation under EnSite 3000 guidance in our hospital were retrospectively summarized. The patients included 93 paroxysmal AF and 43 persistent AF and divided into 4 groups. Paroxysmal AF with CPVAI ablation,n=45, Paroxysmal AF with SL ablation,n=48 and persistent AF with CPVAI ablation, n=18, persistent AF with SL ablation,n=25. The differences of left atrium diameter, ablation time, X-ray exposure time, the success rate and complication were compared among different groups. Results: For 12 months follow-up study, the success rate and complication were similar between 2 ablation methods for treating both Paroxysmal AF and persistent AF patients. For Paroxysmal AF patients, both ablation methods could effectively reduce left atrium diameter,P<0.01. The SL ablation had less procedural time than CPVAI ablation,P<0.01, while the X-ray exposure time was similar between 2 ablation methods. Conclusion: Both CPVAI and SL ablation methods were effective and safe for treating AF patients.

Objective To investigate the use of laparoscopic ultrasound to exclude cystic duct obstruction and its related risk factors in laparoscopic cholecystectomy.Methods The data of 28 patients who underwent laparoscopic cholecystectomy in our department for cystic duct obstruction from February 2008 to April 2010 were analyzed.Subtotal resection of gallbladder and exclusion of cysticduct were carried out when the gallbladder triangle anatomy was not clear.An abdominal drain was used.Results All the patients were cured and there was no bleeding,abdominal infection,or jaundice.On univariate analysis,risk factors for cystic duct obstruction were adhesions in Calot triangle,gallbladder atrophy,acute cholecystitis,cystic duct stone incarceration,gallbladder wall thickening and white bile.Adhesion in Calot triangle,acute cholecystitis and white bile were independent risk factors on multivariate analysis.Conclusion Excluding cystic duct obstruction by laparoscopic ultrasound for patients who underwent laparoscopic cholecystectomy for cystic duct obstruction is safe and effective.

Hepatic artery reconstruction is one of the key procedures in liver transplantation. Accidental dissection of the hepatic artery to be reconstructed caused by donor and recipient factors or surgical factors will disrupt the surgical plan, increase the difficulty of arterial reconstruction, significantly prolong the operation time, increase the risk of postoperative arterial stenosis and thrombosis and probably lead to acute allograft failure, which requires emergency surgical interventions or even secondary liver transplantation. Understanding of how to avoid dissection of the artery to be anastomosed during liver transplantation and corresponding treatment will contribute to preventing the incidence of artery-related complications during liver transplantation and improving clinical prognosis of liver transplant recipients. In this article, the causes, prevention and treatment of hepatic artery dissection and hepatic artery reconstruction in donors and recipients during liver transplantation were illustrated.

Objective:To investigate the factors influencing phytohemagglutinin (PHA) response in the detection of Mycobacterium tuberculosis infection by gamma interferon release assay (IGRA). Methods:A retrospective case-control study was conducted on 360 hospitalized patients who received IGRA in West China Hospital of Sichuan University from January 2019 to December 2021. According to PHA response (IFN-γ level), they were divided into three groups: negative mitogen response group (IFN-γ<2 pg/ml), weak positive mitogen response group (IFN-γ: 2-100 pg/ml), and normal mitogen response group (IFN-γ>400 pg/ml).Results:Immune diseases were independently associated with negative (OR=0.34, 95%CI: 0.17-0.72, P=0.004) and weak positive mitogen responses (OR=0.29, 95%CI: 0.16-0.55, P<0.001). Infections caused by pathogens other than Mycobacterium tuberculosis was independently associated with negative mitogen response (OR=0.266, 95%CI: 0.09-0.83, P=0.023), while immunodeficiency was independently associated with weak positive mitogen response (OR=0.280, 95%CI: 0.12-0.63, P=0.002). Mitogen response was significantly correlated with the levels of albumin and hemoglobin in serum and the counts of neutrophils and lymphocytes ( P<0.001). Conclusions:Immune diseases and immunodeficiency can affect mitogen response. Therefore, clinicians should give attention to mitogen response in the interpretation of IGRA test results to prevent misdiagnosis and underdiagnosis. Besides, to a certain extent, mitogen response can reflect the infection status of hospitalized patients.

Objective To investigate the application value of artery approach in the lower colon region combined with portal vein (PV) resection and allograft vascular grafts in radical pancreaticoduodenectomy for pancreatic cancer combined with vascular invasion.Methods The retrospective descriptive study was conducted.The clinicopathological data of 13 patients with pancreatic cancer involving in PV,splenic vein or junction who were admitted to the Beijing Chao Yang Hospital of Capital Medical University from March 2014 to June 2015 were collected.The superior mesenteric artery (SMA),tumors and soft tissues (including involved vessels) in the right of the celiac trunk were resected after exploring SMA and evaluating resectability of tumors.Patients underwent PV-splenic vein resection and reconstruction with allogenic vein.Observation indicators:(1) surgical situations;(2) postoperative situations;(3) follow-up situation.Follow-up using outpatient examination and telephone interview was performed to detect survival of patients and tumor recurrence and metastasis up to April 2016.Measurement data with normal distribution were represented as (x)±s.Results (1) Surgical situations:13 patients successfully underwent radical pancreaticoduodenectomy via artery approach in the lower colon region combined with PV,splenic vein resection and allograft vascular grafts.Operation time and volume of intraoperative blood loss were respectively (489 ± 31) minutes and (407 ± 96) mL,without intra-and post-operative deaths.(2)Postoperative situations:of 13 patients,3 and 1 patients were respectively complicated with pancreatic fistula (2 in grade A and 1 in grade B) and gastroplegia,and cured by conservative treatment.There was no occurrence of bleeding,intraperitoneal infection,diarrhea,anastomotic stenosis and thrombus.The median duration of postoperative hospital stay was 12 days.Results of postoperative pathological examination:of 13 patients,high-,moderate-and low-differentiated adenocarcinoma was detected in 2,7 and 4 patients respectively.Three patients had negative vascular margin,2 had tunica intima invasion and 8 had tumor cell invasion in vascular adventitia.One,2,6,4 patients were detected in Ⅰ B,Ⅱ A,Ⅱ B and Ⅲ staging,respectively.The negative margin rate by postoperative pathological examination was 11/13.(3) Follow-up situation:13 patients were followed up 10 months postoperatively,with good survival and without tumor recurrence or metastasis.Conclusion The radical pancreaticoduodenectomy via artery approach in the lower colon region combined with PV/SMV resection and allograft vascular grafts is safe and feasible for pancreatic cancer involving in PV,splenic vein or junction,it can also evaluate early resectability of tumors,with good operative efficacy.

Objective:To investigate the effects and mechanism of mitochondrial transcription factor A (TFAM) and cytochrome c oxidase (COX) pathway in the energy production of hypoxic cardiomyocytes of rats regulated by tumor necrosis factor receptor associated protein 1 (TRAP1).Methods:The cardiomyocytes were isolated from 135 neonatal Sprague-Dawley rats (aged 1-3 d) and cultured for the following experiments. (1) Cells were collected and divided into normoxia blank control (NBC) group, hypoxia blank control (HBC) group, hypoxia+ TRAP1 over-expression control (HTOC) group, and hypoxia+ TRAP1 over-expression (HTO) group according to the random number table (the same grouping method below), with 1 bottle in each group. Cells in NBC group were cultured routinely, cells in HBC group were cultured in hypoxic condition for 6 hours after routine culture, cells in HTOC and HTO groups were respectively added with TRAP1 over-expression empty virus vector and TRAP1 over-expression adenovirus vector virus suspension for transfection for 48 hours after routine culture and then cultured in hypoxic condition for 6 hours. The protein expression of TFAM of cells in each group was detected by Western blotting. (2) Cells were collected and divided into NBC, HBC, HTOC, HTO, HTO+ TFAM interference control (HTOTIC), and HTO+ TFAM interference (HTOTI) groups, with 1 well in each group. Cells in the former 4 groups were dealt with the same methods as the corresponding groups in experiment (1). Cells in HTOTIC and HTOTI groups were respectively added with TFAM interference empty virus vector and TFAM interference adenovirus vector virus suspension for transfection for 48 hours, and the other processing methods were the same as those in HTO group. The content of ATP of cells in each group was determined by ATP determination kit and microplate reader, and the COX activity of cells in each group was determined by COX activity assay kit and microplate reader. (3) Cells were collected and divided into NBC group, normoxia+ sodium azide (NSA) group, HBC group, and hypoxia+ sodium azide (HSA) group, with 1 well in each group. Cells in NBC and HBC groups were respectively dealt with the same methods as the corresponding groups in experiment (1). Cells in NSA and HSA groups were respectively added with 32 nmol sodium azide at 30 min before experiment or hypoxia, and then cells in HSA group were cultured in hypoxic condition for 6 hours. The content of ATP was determined by the same method as above. The above three experiments were repeated for three times. Data were statistically analyzed with one-way analysis of variance and least significant difference test.Results:(1) Compared with that in NBC group, the protein expression of TFAM of cells in HBC group was significantly decreased ( P<0.01). Compared with that in HBC group or HTOC group, the protein expression of TFAM of cells in HTO group was significantly increased ( P<0.01). (2) Compared with 0.552±0.041 and 1.99±0.15 in NBC group, the COX activity (0.270±0.044) and ATP content (1.09±0.11) of cells in HBC group were significantly decreased ( P<0.01). Compared with 0.269±0.042 and 1.17±0.12 in HBC group and those in HTOC group, the COX activity (0.412±0.032 and 0.404±0.016) and ATP content (1.75±0.06 and 1.69±0.07) of cells in HTO and HTOTIC groups were significantly increased ( P<0.01). Compared with those in HTO and HTOTIC groups, the COX activity (0.261±0.036) and ATP content (1.23±0.07) of cells in HTOTI group were significantly decreased ( P<0.01). (3) Compared with that in NBC group, the ATP content of cells in NSA and NBC groups was significantly decreased ( P<0.01). Compared with that in HBC group, the ATP content of cells in HSA group was significantly decreased ( P<0.01). Conclusions:TRAP1 can increase the COX activity of cardiomyocytes by raising the expression of TFAM, and finally alleviate the impairment in energy production of cardiomyocytes caused by hypoxia.

Objective:To investigate the effect and mechanism of periodontal ligament stem cell (PDLSC) from inflammatory environment on the secretion of interleukin-1β (IL-1β) by macrophages.Methods:PDLSCs were pretreated with lipopolysaccharide (LPS) in order to simulate the inflammatory environment. Human monocyte cell line (THP-1) cells were treated with conditioned media collected from healthy and inflammatory PDLSCs respectively and divided into conditioned medium of health PDLSC (CM-H) group and conditioned medium of LPS-PDLSC (CM-LPS) group. After 24 h of co-culture, the condition media were abandoned and THP-1 cells were then cultured for another 24 h. The expression of IL-1β in THP-1 cells supernatant was detected by enzyme-linked immunosorbent assay (ELISA). Quantitative real time-PCR (qRT-PCR) was used to detect the expression of glucose regulated protein 78 (GRP78), activating transcription factor-6 (ATF6), inositol requiring enzyme 1 (IRE1), protein kinase R-like endoplasmic reticulum kinase (PERK), CCAAT enhancer binding protein homologous protein (CHOP), activating transcription factor-4 (ATF4) and X box binding protein 1 spliced (XBP1s), which were all related with endoplasmic reticulum stress (ERS), in THP-1 cells. The expressions of proteins GRP78 and CHOP were detected by Western blotting. Furthermore, THP-1 cells, which pretreated with ER inhibitor 4-phenylbutyrate (4-PBA) for intervention experiments were grouped by various concentrations of 4-PBA including groups 0 (control group), 1, 10 and 20 mmol/L and treated with condition medium of inflammatory PDLSC. ELISA was used to detect IL-1β expression and qRT-PCR to detect expression of ERS related genes.Results:ELISA results showed that the expression of IL-1β in THP-1 cells of group CM-LPS [(31.35±2.11) ng/L] was significantly higher than group CM-H [(8.19±1.51) ng/L] ( t=12.60, P<0.01). qRT-PCR results showed that the relative expressions of GRP78, ATF6, IRE1, PERK, CHOP, ATF4 and XBP1s genes in THP-1 cells of group CM-LPS (1.782±0.070, 1.387±0.204, 1.404±0.119, 1.777±0.187, 1.325±0.156, 1.295±0.066 and 1.137±0.149, respectively) were significantly higher than those in group CM-H ( P<0.05). In the 4-PBA intervention experiment, compared with group 0 mmol/L, the expressions of GRP78, IRE-1, ATF-6, PERK and CHOP were significantly lower in group 1, 10 and 20 mmol/L ( P<0.05). Moreover, compared with control group [(31.23±1.98) ng/L], the expression of IL-1β in THP-1 cells were significantly lower in group 10 mmol/L [(21.20±0.37) ng/L] and group 20 mmol/L [(23.85±1.80) ng/L] ( P<0.05) with ERS inhibited. Conclusions:PDLSC from inflammatory environment could promote IL-1β secretion of macrophages through upregulating macrophages ERS.

Objective:To investigate the effect of low dose lipopolysaccharide (LPS)-treated human periodontal ligament stem cells (HPDLSC) on the expression of macrophage pro-inflammatory factors and the mechanism involved.Methods:The primary HPDLSCs were obtained from healthy third molar periodontal ligament tissue. Phosphate buffer saline (PBS), 100 μg/L or 10 mg/L of LPS were used to treat HPDLSCs for 48 h, and their conditioned media were respectively co-cultured with THP-1-derived macrophages for 48 h. The corresponding experimental groups were PBS-treated HPDLSC-derived conditioned medium (CM-C) group, low dose LPS-treated HPDLSC-derived conditioned medium (CM-L) group, and high dose LPS-treated HPDLSC-derived conditioned medium (CM-H) group. Quantitative real-time PCR was performed to explore the mRNA expressions of macrophage interleukin (IL)-6, IL-8, IL-12, tumor necrosis factor-α (TNF-α) in the CM-C, CM-L and CM-H groups, and the expressions of nuclear factor (erythroid-derived 2)-like 2 (NRF2), glutamate-cysteine ligase catalytic subunit (GCLC), NAD(P)H quinone dehydrogenase 1 (NQO1) and heme oxygenase 1 (HO-1) in the CM-C and CM-L groups. Meanwhile, Western blotting was used to detect the change of nuclear and cytoplasmic NRF2 and the levels of GCLC and HO-1 in the CM-C and CM-L groups. The 2′, 7′-dichlorofluorescein probe was adopted to detect the reactive oxygen species (ROS) levels of macrophages in the CM-C and CM-L groups and the data were characterized by the mean fluorescent intensity (MFI).Results:The mRNA expressions of macrophage pro-inflammatory factors IL-6, IL-8, IL-12 and TNF-α in the CM-H group (2.332±0.594, 3.601±0.639, 2.120±0.677 and 2.468±0.236) were significantly upregulated compared with those in the CM-C group (1.000±0.321, 1.000±0.151, 1.000±0.059 and 1.000±0.095) ( P<0.05); while the relative mRNA levels of IL-6, IL-12 and TNF-α in the CM-L group (0.056±0.002, 0.215±0.024 and 0.567±0.071) were much lower than those in the CM-C group (1.000±0.209, 1.000±0.220 and 1.000±0.220) ( P<0.05). At the mRNA level, the expression of NRF2 was significantly increased in the CM-L group (1.864±0.198) compared with that in the CM-C group (1.000±0.094) ( P<0.05). At the protein level, the cytoplasmic NRF2 and nuclear NRF2 were increased in CM-L group (1.175±0.104 and 1.308±0.082) compared with those in the CM-C group (1.000±0.025 and 1.000±0.049) ( P<0.05). Furthermore, the antioxidative genes, i.e. GCLC and NQO1, localized in NRF2 downstream, were significantly upregulated in the CM-L group (1.786±0.278 and 1.444±0.078) compared with the CM-C group (1.000±0.139 and 1.000±0.226) ( P<0.05). The protein levels of GCLC and HO-1 were augmented in the CM-L group (1.159±0.036 and 1.412±0.075) in contrast with those in the CM-C group (1.000±0.050 and 1.000±0.013) ( P<0.05). In addition, the MFI in the CM-L group (123 419±1 302) was significantly lower than that in the CM-C group (139 193±1 241) ( P<0.05). Conclusions:Low-dose LPS-treated HPDLSCs could regulate oxidative stress response through activating the NRF2 signaling pathway of macrophages and further downregulating the expressions of macrophage pro-inflammatory factors.

OBJECTIVES: Cough variant asthma (CVA) is the main cause of obstinate cough. This study aimed to observe the therapeutic effect of Xiaochuan pill on CVA in a rat model, and to explore the mechanisms. METHODS: The rats were sensitized and challenged with 4% ovaibumin (OA) and 2% Al(OH) to establish the CVA models. They were treated with Xiaochuan pill (at the dose of 0.9, 1.8, 3.6 g/kg) or montelukast sodium once a day for 14 days. After 7 and 14 days of intervention, 5 and 10 rats were randomly selected from each group to collect bronchoalveolar lavage fluid (BALF), trachea, and lungs. The number of white blood cells (WBC) and eosinophils (EOS), and the levels of IL-1β, TNF- α, and IFN-γ in BALF were detected. Histopathological examination of lung tissue was performed to observe the histomorphological changes. The expressions of TLR4, MyD88, NF-κBp65, and p-p65 in lung tissue were detected by Western blotting. RESULTS: The numbers of WBC and EOS in BALF of CVA rats were significantly decreased by Xiaochuan pill (<0.05 or <0.01). The hyperplasia of tracheal, bronchial mucosa and the infiltration of inflammatory cells in lung were alleviated obviously. After 14 d of intervention, high dose of Xiaochuan pill significantly increased the level of IFN- γ (<0.01), reduced the levels of IL-1β (<0.05) and TNF-α (<0.05), and decreased the expressions of TLR4, MyD88, p65, and p-p65 (<0.05 or <0.01). CONCLUSIONS Xiaochuan pill exerts the significant therapeutic effect on obstinate cough in rats. The mechanism of action may be related to the regulation of TLR4-MyD88-NF-κBp65 signaling pathway as well as the inflammation and immune response.
Animals ; Cough ; Myeloid Differentiation Factor 88 ; NF-kappa B ; Rats ; Signal Transduction ; Toll-Like Receptor 4 ; Tumor Necrosis Factor-alpha