Ischemic stroke is a common neurological disorder that can lead to neuronal death and neurological dysfunction. Microglia is the main immune cells in the central nervous system, involved in post-stroke inflammation and tissue repair. Triggering receptor expressed on myeloid cells 2 (TREM2), a receptor expressed on the surface of microglia, plays a multifaceted role in neuronal survival and nerve repair after ischemic stroke, including promoting the phagocytosis of microglia, inhibiting excessive inflammatory response, maintaining the proliferation and survival of microglia, protecting neurons from damage, and promoting the recovery of nerve function. Therefore, elucidating the immunoregulatory mechanism of TREM2 on microglia after cerebral ischemia is of great significance for exploring new therapeutic directions for ischemic stroke.

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.

Objective To investigate the feasibility of urethral prefabrication with vaginal mucosa in female-to-male transsexuals and to compare the urologic complications after penile reconstruction in female-to-male transsexuals between prefabrication group and forearm group.Methods Prefabrication of the neourethra with tubed vaginal mucosal graft was performed for 22 female-to-male transsexual patients from 2007 to 2016,while radial forearm flap,known as the traditional tube-within-tube method,was used to construct the neourethra for other 31 cases.Results All of the reconstructed penises survived completely and allowed the patients to urinate while standing in the prefabrication group.Phalloplasty by using the vaginal mucosal graft for urethroplasty significantly decreased the donor scar,the duration of the second operation and the incidence rates of urologic strictures,comparing with the forearm group (P＜0.05).Conclusions It is reliable to use the prefabrication of the neourethra with tubed vaginal mucosal graft in phalloplasty female-to-male transsexual patients.

Purpose To investigate the relationship between maximum standardized uptake value (SUVmax) of 18F-FDG PET/CT and clinical features of tongue squamous cell carcinoma (TSCC),in order to provide better PET/CT results for clinical guide.Materials and Methods Fifty-two patients with pathologically confirmed TSCC who accepting PET/CT examination before surgery were retrospectively analyzed.Single-factor analysis and multiple regression analysis were conducted on possible factors influencing primary tumor SUVmax,including gender,age,smoking history,tumor location,tumor size,TNM stage,T stage and N stage.Results Single-analysis showed that SUVmax was correlated with gender,tumor location,tumor size,TNM stage,T stage and N stage (P＜0.05),and was not correlated with tumor differentiation,smoking history and age (P＞0.05).Multivariate linear regression analysis showed that gender,tumor location,tumor size,T stage and N stage were independent influencing factors of primary tumor SUVmax (P＜0.05).Primary tumor SUVmax had predictive value for lymph node metastasis.When the cutoff value was 6.57,the diagnostic efficiency was the highest,the sensitivity was 79.2％ and the specificity was 85.7％.Conclusion TSCC 18F-FDG PET/CT SUVmax is higher among male patients with tongue base tumor location,larger tumor size and lymph node metastasis.Primary tumor SUVmax is of important significance in predicting lymph node metastasis.

Objective To investigate the characteristics of 18F-FDG PET/CT imaging in pulmonary sclerosis pneumocytoma (PSP).Methods The clinical and PET/CT data of 16 patients with pathologically proved PSP were retrospectively analyzed.The location,shape,size,internal and external edge of the lesion,as well as the metabolism of the lesions were observed.The mean retention index (RI) was calculated in 6 patients with 18F-FDG PET/CT dual phase imaging.The difference of SUVmax between early and delayed phase were compared.And the correlation between the diameter of lesions and SUVmax were analyzed.Results There were 16 lesions in all 16 patients,including 7 cases located at right lung and 9 located at left lung.The lesions were round with the diameter of (1.97-4-0.61)cm.The uniform density were observed with the CT value of (29.87±4.71)HU.And there was no cystic degeneration and necrosis.Calcification was found in 5 lesions.The edge of 14 lesions was smooth,and the edge of another 2 lesions showed short spicular sign.Two lesions showed visible edges of ground glass opacity.There were 12 lesions with vascular welt sign and 3 lesions with air crescent sign.The SUVmax value of PSP was 2.71 ± 2.13.There was no significant difference between the early SUVmax (2.44±1.57) and delayed SUVmax (2.74±1.83) in patients with dual phase imaging (t=2.09,P＞0.05).RI was (7.23±10.29)％.There was no correlation between PSH diameter and SUVmax(r=0.188,P＞0.05).Conclusion Most of PSP showed solitary pulmonary nodules in PET/CT imaging.The radioactive distribution was mild and moderate increase.The vascular welt sign,air crescent sign and the surrounding ground glass opacity are the references findings of PSP.

Objective To reduce the incidence rate of hospital-wide indwelling needle catheter blockage through a nursing project.Methods We set up a nursing project team,conducted a cross-sectional survey to know the using status of hospital-wide catheter,and analyzed the reasons of catheter blockage by root cause analysis method. "Reducing the incidence rate of hospital-wide indwelling needle catheter blockage" was established as the theme;improvement activities of nursing project were planned and developed for six months;and improvement methods were developed and implemented.Results The incidence rate of hospital-wide indwelling needle catheter blockage reduced to 3.6% from 8.1% (χ2=752.177,P<0.01),which reaches the goal of controlling the incidence rate of catheter blockage below 4%,and continuously decreases month by month. After implementation,the score of knowledge questionnaire about indwelling needle catheter blockage of nurses in the wards increased from (78.23±7.14) to (96.4±1.48) (t=54.735,P<0.01).Conclusions The nursing project obviously improves the degree of mastering the knowledge about preventing blockage,and reduces the incidence rate of hospital-wide indwelling needle catheter blockage.

Panax ginseng is a precious medicinal herb both in China and abroad with high medicinal and economic value.Ginseng disease has been recognized as the main factor restricting its application.Modern molecular biology for the disease resistance of ginseng promoted the development of new methods.In this study,it was found that the antimicrobial proteins of ginseng involved lipid transfer protein,cyclophilin,defensins,PR-4 and PR-10,showing inhibitory effects on various pathogens.These findings provided a reference for the control of ginseng diseases.

[ 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 study the microstructural characteristics of white matter fiber tracts of visual pathways using diffusion tensor imaging (DTI)in the patients with primary glaucoma.Methods Twenty patients with primary bilateral glaucoma and 30 nor-mal adults were recruited and scanned with visual pathways magnetic resonance imaging (MRI)and DTI,obtaining T1 FLAIR ima-ges and directionally encoded color (DEC)images.On DEC images,ROIs were put in the anterior,middle and posterior part of bi-lateral optic nerve,optic tract and optic radiation,to calculate the average FA and MD value of each region.Results T1 FLAIR and DEC images could clearly show the optic nerve,optic tract and optic radiation.In patients with glaucoma,the FA values of optic nerve,optic tract and optic radiation were:0.372±0.040,0.340±0.036,0.31 5±0.026,while the MD values were:1.760±0.1 1, 1.831±0.09,1.927±0.10.The FA values of optic nerve,optic tract and optic radiation of normal adults were:0.538 ±0.073, 0.460±0.082,0.455±0.083,whilst the MD values were:0.995 ±0.27,1.312 ±0.40,1.347 ±0.37.Compared with the normal controls,FA values of all parts are deceased whereas the MD values of all parts are increased in patients.Conclusion Glaucoma is a central nervous system disease involving the whole visual pathways.Use DTI can evaluate the microstructural characteristics of white matter fiber tracts of visual pathways in the patients with glaucoma.