Objective To study the effect of caspase-3 on the primary cultured mice cortical neuron w ith herpes simplex virus type 1 (HSV-1) infection. Methods A spectrophotometer was used to de tect the absorbance units at 405 nm of the caspase-3 substrate,the chrom o phore p-nitroanilide(pNA ).Comparing with the normal group, virus group, s orbital group,and sorbital group with virus in the absorbance units of chromophore substrate.The changes in inner mitochondrial membrane p otential of the different group cells were analyzed with flow cytometry. Results The released chromphore was measured by determining the absorbance at 405 nm. N group is 0.394 2, V group is 0.240 8, NS group is 0.539 2, VS gr oup is 0.465 0,the control C1 group is 0.290 0, C2 group is 0.235 0 and all groups were analyzed by statistic (P

Objective To investigate the effects of Rhizoma Alismatis extracts on oxidative stress induced by cerebral ischemia-reperfusion injury in rats,and to explore its protective mechanism in cerebral ischemia-reperfusion injury.Methods A total of 60 male SD rats were randomly divided into sham operation group,model group,Alisma orientalis group and Nimodipine positive control group (n=15,each).Cerebral ischemia-reperfusion injury model was prepared by suture method after 14 days of intragastric administration.After 24 hours,scores of neurological dysfunction,the infarct size,the water content of the brain,the malondialdehyde (MDA),superoxide dismutase (SOD),nitric oxide (NO) levels in serum and brain tissues,and the activity of inducible NO synthase (iNOS)were detected.Results As compared with the model group,Alisma orientalis group showed that the scores of neurological dysfunction,cerebral water content,cerebral infarction size,contents of MDA and NO,and the activity of iNOS were significantly reduced,and the activity of SOD was significantly increased in respectively [(2.21 ± 0.38) vs.(2.78 ± 0.43),(81.18 ± 2.09)％ vs.(88.33±4.15)％,(0.26±0.07) ％ vs.(0.35±0.04)％,(5.92±1.64) μmol/L vs.(8.21±1.47)μmol/L,(115.48±18.65) mU/L vs.(75.52±20.78) mU/L,(28.23±4.32) μmol/L vs.(41.73±3.85) μmol/L,(15.31±1.68) mU/L vs.(23.49±3.53) mU/L,(5.41±0.68) μmol/L vs.(7.58±1.49) μmol/L,(168.57±10.65) mU/L vs.(150.11±13.62) mU/L,(14.37±0.77) μmol/L vs.(22.08±1.57) μmol/L,(9.83±0.75) mU/L vs.(13.28±1.84) mU/L,respectively,all P＜0.05]Conclusions Alisma orientalis extract has the protective effect on focal cerebral ischemia reperfusion injury,and the mechanism may be related to antioxidant and scavenging free radicals.

OBJECTIVETo observe the effect of HSV-1 infection on the viability of the cultured chicken embryo telencephalon neurons in vitro.

METHODSPrimary culture model of chicken embryo of telencephalon neurons was established. Herpes simplex virus type 1 (HSV-1) was propagated in Vero cells and viral titer was measured by plaque forming method. Inverted microscope, transmission electron microscope, MTT cell viability and DNA agarose gels electrophoresis were used to evaluate the effect of HSV-1 infection on the neurons.

RESULTSAt 24, 48, 72 hours post infection the viability of the neurons decreased by 25%, 56%, 97% as compared with the control group. Morphological changes and DNA agarose gel electrophoresis showed a necrotic effect.

CONCLUSIONSHSV-1 infection induced a remarkable decrease on the viability of neurons by necrosis rather than apoptosis.

Animals ; Cercopithecus aethiops ; Chick Embryo ; Herpes Simplex ; pathology ; virology ; Herpesvirus 1, Human ; In Vitro Techniques ; Necrosis ; Neurons ; pathology ; Telencephalon ; cytology ; Vero Cells ; Virus Cultivation

Objective:To investigate the role of mitofusion 2 (Mfn2) in high glucose (HG)-induced endoplasmic reticulum stress (ERS) and apoptosis of podocytes.Methods:(1) Streptozocin was used to induce a diabetes mellitus (DM) rat model. Renal histopathological changes in rats were observed by HE staining. Expression of Mfn2 and CCAAT/enhancer-binding protein homologous protein (CHOP) in glomeruli was observed by immunohistochemistry. Protein levels of Mfn2, protein kinase RNA-like ER kinase (PERK), phospho(p)-PERK, and CHOP in glomeruli were analyzed by Western blotting. (2) Conditionally immortalized human podocytes (HPC) cultured in vitro were divided into control, mannitol (MA) and HG groups. Expression of Mfn2 was observed by immunofluorescence. Protein levels of Mfn2, p-PERK, PERK and CHOP in HPC were analyzed by Western blotting. Podocyte apoptosis in each group was evaluated by flow cytometry with AnnexinⅤ-PE/7AAD double staining method. (3) HPC were divided into control, HG, HG+Mfn2-Myc plasmid-transfected and HG+control plasmid-transfected groups. Protein levels of Mfn2, p-PERK, PERK and CHOP in HPC were analyzed by Western blotting. Expression of CHOP was observed by immunofluorescence. Mitochondrial membrane potential in each group was observed by mitochondrial membrane potential assay kit with JC-1. Podocyte apoptosis in each group was evaluated by flow cytometry with AnnexinⅤ-PE/7AAD double staining method. Results:(1) Compared with the control group, the glomerular mesangial matrix of the DM group rats was significantly proliferated, and the expression of Mfn2 was down-regulated with the expression of ERS-related proteins p-PERK/PERK and CHOP up-regulated (all P<0.05). (2) Compared with the control group, Mfn2 was down-regulated and p-PERK/PERK and CHOP were up-regulated in HPC of HG group (all P<0.05). Apoptosis of HPC was also increased in HG group. There was no significant difference in the above indicators between the control group and the mannitol group (all P>0.05). (3) Compared with the HG group, mitochondrial membrane potential of HPC was alleviated and apoptosis of HPC was decreased in HG+Mfn2-Myc plasmid-transfected group ( P<0.05). P-PERK/PERK and CHOP were down-regulated in HG+Mfn2-Myc plasmid-transfected group (both P<0.05). There was no significant difference in the above indicators between the HG group and the HG+control plasmid-transfected group (all P>0.05). Conclusions:Mfn2 down-regulation in HG-stimulated podocytes may induce ERS to increase apoptosis of podocytes. Up-regulation of Mfn2 can alleviate the HG-induced ERS and apoptosis in podocytes.

Objective To explore the distribution characteristics and function of peripheral regulatory T cells (CD4+CD25+Foxp3+T cells) in patients with systemic lupus erythematosus (SLE).In addition,we analyzed the relationship between peripheral regulatory T cells and organ damage and the influence of different treatment regimens on them.Methods Two hundred and six SLE patients and 38 healthy volunteers were enrolled,which included 12 patients with untreated new-onset lupus,11 patients with drug withdrawal more than six months and 183 patients with treatments.Phenotypic and functional analysis of peripheral blood CD4+CD25+Foxp3+T cells were performed by flow cytometry.The correlations of CD4+CD25+Foxp3+ T cells with disease activity,organ involvement were analyzed.Thealtered frequency of CD4+CD25 +Foxp3+T cells under different treatment regimens was compared.Statistical Package form Soci-science (SPSS) 21.0 software was used for data analysis,Student's t test,one-way ANOVA,Mann-Whitney T test,Kruskal-Wallis test,Chi-square test,Simple linear correlation analysis was used.Results CD4 +CD25 +Foxp3 + T cells were significantly increased inactive SLE patients [1 1.9％ (9.3％,16.0％),mean difference =104.71,P＜0.01] and inactive SLE patients [11.0％(7.7％,14.7％),mean difference=86.10,P＜0.01] compared with healthy controls [6.1％(5.3％,7.4％)].CD4+CD25+Foxp3+T cellsshowed sign-ificantly positive correlations with SLEDAI-2K (r=0.191,P＜0.05),dsDNA (r=0.262,P＜0.05),ESR (r=0.208,P＜0.05) and lgG (r=0.163,P＜0.05),and significantly negatively correlated with complementC3 (r=-0.201,P＜0.05) and C4 (r=-0.227,P＜0.05).Compared with patients without organ damage (Occult lupus),the CD4+CD25+Foxp3+T cells were increased in SLE patients with organ damage,especially those with skin involvement [10.9％(7.8％,13.1％),mean difference=56.93,P＜0.05] and renal involvement [12.1％(9.1％,16.0％),mean difference=77.26,P＜0.05].The proportion of CD4+CD25+Foxp3+T cells had no significant difference between SLE patients with treatments and patients with untreated new-onset lupus.The expressions of CTLA-4 [(53±15)％,t=7.04,P＜0.01],GITR [(42±19)％,t=2.64,P＜0.01] and ICOS [(28±9)％,t=4.27,P＜0.01] on CD4+CD25+Foxp3+T cells were significantly lower in SLE patients than in healthy controls [CTLA-4 (71±4)％,GITR (53±10)％ and ICOS (41±6)％].IL-17 synthesized by CD4+CD25+Foxp3+T cells in SLE patients [3.0％(1.8％,3.9％)] was significantly higher than that in healthy controls [1.0％(0.7％,1.2％),Z=-4.40,P＜0.01].Conclusion The peripheral regulatory T cells are significantly increased in SLE patients and correlate with disease activity and organ damage.However,their inhibitory function is defective and they have more pro-inflammatory character-istics.

Objective:To evaluate the effect of 2-aminoethoxydiphenyl-borate (2-APB) on cardiac dysfunction in arsenic poisoning rats by stratified strain technique.Methods:Thirty-two 12-week-old SD rats were randomly divided into control group ( n=8) and arsenic exposure group ( n=24). After 12 weeks of arsenic exposure, arsenic exposure group was divided into three groups: natural recovery group ( n=8), low-dose intervention group (n=8) and high-dose intervention group ( n=8). The rats were treated with 2-APB for 21 days. After the last administration, the routine parameters and the layered strain parameters were measured by ultrasonic instrument. Then the rats were killed and their blood and myocardial samples were obtained. The levels of serum creatine kinase isoenzyme (CK-MB) and lactate dehydrogenase (LDH) were tested, and the morphological changes of cardiomyocytes were observed by HE staining. Results:Left ventricular ejection fraction (LVEF), left ventricular short axis shortening (LVFS), global circumferential strain of subendocardial myocardium (GCS-endo), global circumferential strain of middle myocardium (GCS-mid), global circumferential strain of subepicardial myocardium (GCS-epi) and circumferential strain rate of left ventricular segments (SrC) in the natural recovery group were lower than those in the control group ( P<0.05), while serum CK-MB and LDH were higher than those in the control group ( P<0.05). Some ultrasonic parameters and biochemical indexes in the low and high dose intervention groups were improved in varying degrees compared with the natural recovery group ( P<0.05). The correlation between GCS-endo and CK-MB was the highest ( r=-0.931, P<0.05). Myocardial HE staining showed that the degree of myocardial cell swelling and necrosis were reduced, and erythrocyte exudation was reduced in the low and high dose intervention groups compared with the natural recovery group. Conclusions:The stratified strain technique can be used to evaluate the protective effect of 2-APB on cardiac dysfunction in arsenic poisoning rats, and GCS-endo may be one of its sensitive indexes.

Objective:To establish silicone cartilage models of donor-sites for the microtia patients by using digital technology, and to explore the application of surgical simulation in auricular reconstruction.Methods:From June 2018 to October 2019, 19 congenital microtia patients underwent thoracic CT scans and following three-dimensional costal cartilage imaging with Mimics software at the Nanfang Hospital, Southern Medical University. Among these patients, 16 were males and 3 were females. The mean age of patients was 16 years (range 8 to 35 years). Silicon cartilage models were produced by 3D printing and used for surgical planning and preoperative simulation in ear framework fabrication. Cartilaginous framework was sculptured according to the simulation during operation. Patients were followed up for a minimum of six months to evaluate the size, outline, height and auriculocephalic angle of the reconstructed ear. The satisfactory outcomes of the patients were scored according to a 5-point Likert scale.Results:All the patients received the surgical simulation and sculpture training with silicone cartilage models before operation. Auricular reconstruction was completed successfully according to the simulation. The duration of sculpture was shortened to 1-1.5 hours. There were no serious complications, such as hematoma, inflammation, skin necrosis and framework exposure. The contour of reconstructed ear was natural and clear over a 6 months follow-up, and all the patients were satisfied with their surgical outcomes.Conclusions:With the application of digital technology and silicone cartilage models by 3D printing to the surgical planning and training in microtia patients, patient-specific framework is fabricated with precisely assembling, which not only shortens the operation time, but also provides the unexperienced surgeons with a safe and effective training of ear framework fabrication.

Objective:To construct a set of weighted trigger index system for dynamic adjustment of medical service price,so as to provide references and basis for price adjustment of complex medical service projects in Tangshan.Methods:The Delphi expert consultation method was used to demonstrate and determine the index system,and the difference comparison method and product method were used to determine the weight coefficient for each index.Results:The trigger index system of dynamic adjustment of medical service price including 4 first-level indicators,10 second-level indicators and 29 third-level indicators,and the weight coefficient of each indicator was given.Conclusion:The trigger index system of dynamic adjustment of medical service price in Tangshan has high scientificity and reliability,which is helpful for Tangshan to establish a sensitive and moderate price dynamic adjustment mechanism.Considering the affordability of all parties in the price adjustment of medical services in Tangshan,it is conducive to strengthening the total price control and structural adjustment of medical services in Tangshan.

Objective: To investigate the role of peroxisome proliferator-activated-receptor-γ coactivator-1α (PGC-1α) in carotid atherosclerosis due to human cytomegalovirus (HCMV) infection. Methods: Thirty-three samples of carotid arterial sclerosis plaques after carotid endarterectomy (CEA) were collected in the experiment group, and 12 pieces of normal intracranial arteries were collected in the control group. The plaques of carotid artery were studied using in situ hybridization (ISH) and immunohistochemistry (IHC) for the role of PGC-1α in HCMV related atherosclerosis. HCMV immediate early (IE) gene of two groups was detected using ISH, and the expression of PGC-1α using IHC. Results: HCMV IE gene was positive 72.7% in the atherosclerotic plague group, while 16.6% in the control group, there was significant difference between the two groups (P<0.05). As to PGC-1α expression, in the atherosclerosis group, 8/33(24.2%) were strongly positive (+ + + ), 9/33(27.2%) were positive (+ + ), 6/33(18.2%) were weakly positive (+ ), and 10/33(30.3%) were negative (-). While in the control group, 1/12(8.3%) was strongly positive (+ + + ), 2/12(16.7%) were positive (+ + ), and 1/12 (8.3%) was weakly positive (+ ), and 8/12 (66.7%) were negative (-). The differences between the two groups were statistically significant (P<0.05). Conclusions HCMV immediate early gene and PGC-1α were both related to athersclerotic plaque formation, suggesting that PGC-1α may play an important role in as the plaque formation due to HCMV infection.

Objective: To investigate the role of the activation and oxidative stress of cultured human umbilical vein endothelial cells (HUVEC) after HCMV infection. Methods: HUVECs were divided into four groups: control, HCMV(+ ), after HCMV AD169 infection, and the supernatant of the culture was extracted, and the cells were lysed. The levels of vascular cellular adhesion molecule-1 (VCAM-1) in HUVEC were measured by real-time PCR. And the content of nitrogen monoxide (NO) of the supernatant was detected by nitrate reductasemethod accordingly. Results: Twenty-four hours after infection, the mRNA expression of VCAM-1 in HUVECs of HCMV infected group increased obviously compared to control, and NO quantity increased accordingly and time-dependently. There was significant difference between groups(P<0.01). Conclusions HCMV increases the mRNA expression of VCAM-1 and quantity of NO, which may contribute to the formation of AS.