AIM:To investigate the distribution of protein Z intron FG79A polymorphisms in Chinese and the associations with coronary artery disease(CAD).METHODS:148 patients were performed selected coronary angiography and more than one major coronary vessel with at least 50% stenosis was defined as CAD.The control group consisted of 147 subjects.The protein Z intron FG79A polymorphisms were studied by polymerase chain reaction and restriction fragment length polymorphism and parts of PCR products were sequenced.RESULTS:Protein Z intron FG79A polymorphisms were first recovered in Chinese,and the frequencies of G and A alleles were 44.24% and 55.76%,respectively.The frequencies of two alleles were not significantly different between patients and controls.There was no significant difference in protein Z intron FG79A genotype distribution among patients with one,two or three stenosed vessels.No significant difference was found among the frequencies of the three genotypes between both acute coronary syndrome(ACS)and non-ACS.CONCLUSION:Protein Z intron FG79A polymorphisms are present in Chinese.Protein Z intron FG79A polymorphisms were not associated with CAD and ACS.

Aim To define the effect of genistein (GST) on ceruloplasmin (CP) expression in cerebral cortex of focal cerebral ischemia male rats.Methods The effect of GST on CP expression were investigated by means of immunohistochemistry and Western blot in cerebral cortex of focal cerebral ischemia male rats.Results The expression of CP was time coursed during brain ischemia. According to the sham group,CP expression decreased in 6 h-,12 h-and 24 h-ischemia groups,and then increased gradually in 48 h-and 72 h-ischemia rats. GST weakened the ischemia-induced CP change.Pretreatment with an antiestrogen tamoxifen (TAM) could completely abolish the effect of GST.Conclusion The present results indicate that GST may inhibit the time coursed expression changes of CP and then provide neuroprotective effects by inhibiting iron related oxidative stress during brain ischemia via estrogen receptor.

[ ABSTRACT] AIM:To investigate the effects of tanshinone IIA ( Tan IIA) on proliferation, apoptosis and its mo-lecular mechanism in human hepatoma HepG2 cells under hypoxic condition.METHODS:Hypoxia model was established by treatment with cobalt chloride ( CoCl2 ) .The cells were divided into normoxia control group, hypoxia control group and hypoxia combined at different concentrations of Tan IIA groups.After HepG2 cells were incubated with different concentra-tions of Tan IIA (0.5, 1.0, 2.0, 5.0 and 10.0 mg/L) for 24 h, 48 h and 72 h under hypoxic condition, the cell prolifer-ation was determined by MTT assay.After Tan IIA was added to the media at different concentrations for 24 h and 48 h, the apoptotic cells were observed by Hoechst 33258 staining.The protein levels of hypoxia-inducible factor 1 alpha (HIF-1α) , vascular endothelial growth factor ( VEGF) and wild-type P53 were detected by Western blotting after cultured with different concentrations of Tan IIA for 48 h.RESULTS:Tan IIA inhibited the proliferation of HepG2 cells in a dose-and time-dependent manner.Tan IIA induced the typical morphology of apoptotic cells and increased the apoptotic rate in a dose-and time-dependent manner after treatment with 1.0 mg/L~5.0 mg/L for 24 h and 48 h under hypoxic condition. The protein levels of HIF-1αand VEGF were weakly expressed in HepG2 cells under normoxia but up-regulated after incu-bated under hypoxia for 48 h.The protein expression of HIF-1αand VEGF were decreased with the increase in the concen-tration of Tan IIA under hypoxia.The protein expression of wild-type P53 was increased with the increase in the concentra-tions of Tan IIA under hypoxia.CONCLUSION:Tan IIA significantly inhibits the proliferation and induces the apoptosis of human hepatoma HepG2 cells under hypoxia, which may be related to the down-regulation of HIF-1αand VEGF and up-regulation of wild-type P53.

Aim To investigate the mechanism of demethylation on adenosine (ADO )and homocysteine (HCY)-induced apoptosis in human hepatoma HepG2 cells .Methods HepG 2 cells were treated with differ-ent concentrations of ADO (1.0、2.0、4.0 mol · L-1 ) alone or in combination with HCY for 6h,12h and 24h,5-aza-2-deoxycytidine (5-Aza-CdR)as a positive control.Cell viabilities were assessed by CCK8 assay. Cell apoptosis was observed by AnnexinV-FITC/PI staining.The mitochondrial membrane potentials(ΔΨ) were measured by flow cytometry.The mRNA and pro-tein expressions of caspase-3,caspase-8,caspase-9, MDM-2,p53,Cytochrome C,DNMT1,DNMT3a,DN-MT3 b were detected by RT-qPCR and Western blot re-spectively.Results ADO alone or in combination with HCY significantly reduced the viability of HepG2 cells in a dose and time-dependent manner.The apoptotic rates of HepG2 cells after combination treatment with ADO and HCY at 1 .0,2.0,4.0 mol · L-1 for 24 h were (1 8.63 ± 1.25 )%,(29.42 ±2.37 )% and (42.47 ±3.09 )%,compared with the control group (1.30 ±0.82 )%,P <0.01;and the mitochondrial membrane potentials were decreased from 674.15 ± 82.8%(black control group)to (428.38 ±54.5)%, (297.78 ±30.5)%,(74.45 ±5.73)%,P<0.01, respectively.The expressions of caspase-3,caspase-8, caspase-9,MDM-2,p53,Cytochrome C were up-regula-ted and MDM-2 were down-regulated after combination treatment of ADO and HCY.The mRNA expressions of DNMT1 ,DNMT3 a and DNMT3 b were down-regulated after combination treatment with ADO and HCY or 5-Aza-CdR alone.Conclusion Combination treatment of ADO and HCY can cause cellular methylation chan-ges.The effects of demethylation of ADO and HCY may activate p53 gene and mitochondrial pathway, which at last leads to HepG2 cell apoptosis.

Objective To investigate the relationship between inflammation and blood coagulation function in the patients with acute exacerbation of chronic cor pulmonale (AECCP) and discuss the potential mechanism and influence on the patients. Methods The present study was based on 30 healthy controls and 141 cases of AECCP in our hospital from January 2011 to June 2014.Levels of white blood cell (WBC), neutrophil (NEUT), high-sensitivity C-reactive protein (hs-CRP, Complement 3 (C3), prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB) and thrombin time (TT) in the patients were determined. Results Compared with the healthy controls, the patients had higher levels of WBC, NEUT, hs-CRP, PT, APTT, FIB, TT (all P < 0.001) and lower level of C3 (P < 0.001). Significant positive correlations were found between the levels of WBC, NEUT and FIB (r = 0.196 and r = 0.199, both P < 0.05); hs-CRP and APTT, FIB(r = 0.234, P < 0.01 and r = 0.466, P < 0.001); C3 and FIB(r = 0.466, P < 0.001), and significant negative correlations were observed between the levels of C3 and PT, APTT, TT (r=-0.258, P<0.01;r=-0.279, P < 0.01 and r = -0.168, P < 0.05, respectively). Compared with the survival patients, the cases of death had higher levels of WBC and NEUT (both P < 0.01). The area under receiver operating characteristic curve of WBC and NEUT, predicting the prognosis, was 0.666 (95% CI 0.552, 0.780; P < 0.01) and 0.695 (95% CI 0.558, 0.801; P = 0.001) respectively. Conclusions Inflammation and blood coagulation function disorder usually coexist in the patients with AECCP, and are closely associated with the severity. Levels of both WBC and NEUT can be used as prognosis predictors for the patients.

Objective:To investigate the clinical efficacy of microsurgical resection in primary jugular foramen schwannomas (JFSs).Methods:A retrospective analysis was performed; the clinical data of 58 patients with JFSs treated by microsurgery in Department of Neurosurgery, First Affiliated Hospital of Sun Yat-sen University from May 2012 to June 2021 were collected. Seven patients accepted microsurgery via suboccipital retrosigmoid approach, and 51 patients accepted microsurgery via jugular foramen approach. Fifty-three patients were followed up for 4.5 years (ranged from 0.5-8.5 years); follow-ups included Karnofsky performance status (KPS) scores, postoperative complications, and imaging reexaminations. Results:Fifty patients (86.2%) achieved total tumor resection and 8 (13.8%) subtotal resection. The KPS scores at discharge were 68.6±14.9, which were significantly lower than the preoperative KPS scores (77.6±13.5, t=2.452, P=0.017). During the follow-up, 5 patients(9.4%) had tumor recurrence, and 39 patients (73.6%) had improved symptoms after surgery. One patient (1.9%, modified Samii D type) died of cerebellar hemorrhage and swelling after surgery. The main complications included new/aggravated hoarseness (11/53), cerebrospinal fluid leakage (7/53), new/aggravated dysphagia (5/53), and new facial paralysis (4/53). Conclusion:In microsurgical resection of JFSs, short-term symptoms of the lower cranial nerves may be exacerbated, but long-term results are good.

Objective:To investigate the effect of LRRK2G2019S mutation on activation of microglia after iron deprivation and its mechanism.Methods:(1) Microglia were differentiated from human induced pluripotent stem cells (IPSC) with the help of hematopoietic progenitor cells (HPC) and identified by immunofluorescent staining, and α-synuclein (α-syn) A53T mutant protein was obtained by protein purification technology. (2) Microglia were divided into control group, α-syn group, α-syn+ deferoxamine (DFO) group; phosphate buffer solution (PBS), 1 μmol/L purified α-syn A53T mutant protein, 1 μmol/L purified α-syn A53T mutant protein+30 mmol/L DFO were given respectively for 24 h. Fe 2+ concentration was detected by colorimetry, Rab35 protein expression was detected by Western blotting, intracellular reactive oxygen species (ROS) level was detected by flow cytometry, and interleukin-6 ( IL-6), tumor necrosis factor-α ( TNF-α) and transforming growth factor-β ( TGF-β) mRNA expressions were detected by real time-PCR (RT-PCR); microglia culture supernatant (MCS) in the 3 groups were transfered to SH-SY5Y cells, and SH-SY5Y cell apoptosis was detected by flow cytometry. (3) Bidirectional DNA sequencing was used to detect leucine rich repeat kinase 2 ( LRRK2) gene mutations in microglia treated with 1 μmol/L purified α-syn A53T mutant protein. Microglia were divided into control group, α-syn group and α-syn+GSK3357679A group, and treated with corresponding drugs for 24 h, respectively (LRRK2 inhibitor GSK3357679A concentration: 10 nmol/L), and LRRK2 protein expression was detected by Western blotting; microglia were divided into control group, α-syn group, α-syn+GSK3357679A, and α-syn+GSK3357679A+DFO group, and treated with corresponding drugs for 24 h, Rab35 protein expression was detected by Western blotting, intracellular ROS level was detected by flow cytometry, and IL-6, TNF-α and TGF-β mRNA expressions were detected by RT-PCR. (4) Microglia were divided into control group, α-syn group, α-syn+rapamycin (RAPA) group, and treated with corresponding drugs for 24 h (concentration of autophagy inducer RAPA: 50 nmol/L); protein expressions of Rab35, P62 and microtubule-associated protein light chain 3 II (LC3II) were detected by Western blotting; intracellular ROS level was detected by flow cytometry, and IL-6, TNF-α and TGF-β mRNA expressions were detected by RT-PCR. (5) Microglia were divided into control group, α-syn group, and α-syn+Rab35 group, and treated with corresponding drugs for 24 h (concentration of Rab35 overexpressed plasmids: 1 μg/mL); Rab35, P62, and LC3II protein expressions were detected by Western blotting; ROS level was detected by flow cytometry, and IL-6, TNF-α and TGF-β mRNA expressions were detected by RT-PCR. Results:(1) Immunofluorescent staining showed negative neuronal nuclei (NeuN) expression and positive ionized calcium-binding adapter molecule 1 (Iba1) expression in microglia, and high LRRK2 expression; PcDNA3.1-SNCA-A53T expression plasmid was constructed and α-syn A53T mutant protein was purified. (2) The Fe 2+ concentration in α-syn group was significantly higher than that in control group, and the Fe 2+ concentration in α-syn+DFO group was significantly lower than that in α-syn group ( P<0.05); the Rab35 protein and TGF-β mRNA expressions in control group, α-syn group and α-syn+DFO group were decreased successively, while the IL-6 and TNF-α mRNA expressions were increased successively, with significant differences ( P<0.05); ROS level and SH-SY5Y cell apoptosis rate in control group, α-syn group, α-syn+DFO group were increased successively. (3) Bidirectional DNA sequencing showed that the LRRK2G2019S mutation in microglia was the most obvious after α-syn A53T mutant protein stimulation; compared with the control group, the α-syn group had significantly increased LRRK2 protein expression, while the α-syn+GSK3357679A group had significantly decreased LRRK2 protein expression compared with α-syn group ( P<0.05); compared with the control group, the α-syn group had significantly decreased Rab35 protein and TGF-β mRNA expressions, and statistically increased IL-6 and TNF-α mRNA expressions ( P<0.05); compared with α-syn group, the α-syn+GSK3357679A group had significantly increased Rab35 protein and TGF-β mRNA expressions, and statistically decreased IL-6 and TNF-α mRNA expressions ( P<0.05); compared with α-syn+GSK3357679A group, α-syn+GSK3357679A+DFO group had significantly increased IL-6 and TNF-α mRNA expressions, and significantly decreased Rab35 protein and TGF-β mRNA expressions ( P<0.05). The α-syn group had higher ROS level than the control group, the α-syn+GSK3357679A group had lower ROS level than the α-syn group, and the α-syn+GSK3357679A+DFO group had higher ROS level than the α-syn+GSK3357679A group. (4) Compared with the control group, the α-syn group had significantly decreased Rab35 and LC3II protein, and TGF-β mRNA expressions, and significantly increased P62 protein, IL-6 and TNF-α mRNA expressions ( P<0.05); compared with α-syn group, the α-syn+RAPA group had significantly increased Rab35 and LC3II protein, and TGF-β mRNA expressions, and significantly decreased P62 protein, and IL-6 and TNF-α mRNA expressions ( P<0.05); the α-syn group had higher ROS level than the control group and α-syn+RAPA group. (5) Compared with the control group, the α-syn group had significantly decreased Rab35 and LC3II protein, and TGF-β mRNA expressions, and statistically increased P62 protein, and IL-6 and TNF-α mRNA expressions ( P<0.05); compared with the α-syn group, the α-syn+Rab35 group had significantly increased Rab35 and LC3II protein, and TGF-β mRNA expressions, and significantly decreased P62 protein, and IL-6 and TNF-α mRNA expressions ( P<0.05). The α-syn group had higher ROS level than the control group and α-syn+Rab35 group. Conclusion:LRRK2G2019S can induce neuroinflammation by inhibiting Rab35-related autophagy under iron deprivation, and Rab35 is expected to be a key factor in intervening neuroinflammation.