Objective To explore the effect and mechanism of taking sibelium long term for treatment and preventing the attack again of cerebral infartcion.Methods 339 patients with cerebral infarction were divided into 2 groups randomly,namely preventing group(120 cases) and control group(119 cases).The basic treatment in the two groups are the same.The preventing group was taken siblium 10 mg once every evening,at least 6 months.To observe the clinical effect in 1 month and the rate of attack again in 1 year.The two groups were measured on hemorrheology,blood lipid and platelet aggregation rate before and after treatment one year.Results After treatment one month,the significant effective rate in sibelium preventing group is 80%,the total effective rate is 98.3%,but the control group is 54.6% and 74.8%(all P

AIM: To explore the role of plasma endothelin (ET) in patients with acute cerebral infarction (ACI) and its change after nimodipine treatment.METHODS:Sixty-six patients with ACI were randomly divided into 2 groups according to blood pressure:hypertension ACI group 35 patients［M 20,F 15;age (65±　s　11) a］ and pure ACI group 31 patients［M 17,F 14; age (62±10) a］.Plasma ET was measured by radioimmunoassay and compared with 27 healthy individuals.Among them,42 patients（hypertension ACI group 25 patients，pure ACI group 17 patients） with ACI were given nimodipine 4 mg, iv,gtt, qd, for 2 wk and measured ET again.　RESULTS:Plasma ET in 2 groups were (144±42) ng*L-1 and (72±35) ng*L-1 respectively, and that were (94±55) ng*L-1 and (60±37) ng*L-1 respectively after treatment with nimodipine, but was still higher than that in healthy individuals (P<0.01).　CONCLUSION: The level of plasma ET was associated with blood pressure and the severity of disease. ET is closely related with ACI. Nimodipine reduces the plasma ET while nerve function default degree is effectively improved.

Objective To study the clinical efficacy of computer-assisted stereotactic brain transplantation of human retinal pigment epithelium (hRPE) cells into the patients with Parkinson disease (PD). Methods Under the guidance of computed X-ray tomography and magnetic resonance imaging image mergence, 4 × 106 hRPE cells were transplanted into the putamen and ventriculus laterlis of 17 cases of PD by stereotactic surgery. The transplantation sites were contralateral to the side of main symptoms and signs. The curative efficacy were observed at the 7th day, 1st month, and 3rd month after the transplantation. Results The contralateral symptoms were ameliorated continuously after the transplantation. Three months after the surgery, the total effective rate of cell transplantation was 88. 2 %, and 82. 4 % of the cases got significant improvement. The cases that got ipsilateral improvement soon after the surgery gave a total effective rate as high as 88. 2 % at the 3rd month during follow-up period, and 64. 7% among these cases improved significantly. Only a minority of cases had transient dizziness and hemiparesis, but the duration was short. Conclusion The therapy, computer-assisted stereotactic transplantation of hRPE ceils in the treatment of PD, is safe and efficient.

ObjectiveTo explore effect of interleukin-10 (IL-10) on histopathologic scores of severe acute pancreatitis (SAP) and the level of serum amylase (AMY) in rats.Methods48 female and male adult Sprague Dawley rats (200—300g) were randomly allocated into three groups: OPgroup, SAP group and IL-10 group with 16 rats in each group. SAP was made with retrograde ductal infusion of 5% sodium taurocholate solution.ResultsIn SAP and IL-10 group, there were interstitial edema, necrosis, neutrophil infiltration and interstitial hemorrhage of pancreas, more or less. At 6h and 12h after models were made, the pancreatic histopathologic score in IL-10 group (4.00±0.33 and 6.25±0.25) were significantly lower than that in SAP group (6.13±0.35 and 9.50±0.50)(P<0.01). At 6 h after models were made, the serum AMY in IL-10 group was lower than that in SAP group (P<0.05), but at 12 h there were no differences.ConclusionIn earlier period of SAP in rats, IL-10 can lower the serum AMY level, and significantly reduced pancreatic histologic score (edema, inflammation, hemorrhage, and necrosis).

Objective:To observe the inhibition of SARS-CoV-2 spike protein (S-protein) on the proliferation of human retinal pigment epithelium (RPE) cells.Methods:SARS-CoV-2 S-protein gene fragment expression plasmid (p3xflag-S) was constructed and transfected into human RPE, HEK293 cells. DNA sequencing was used for identification, and the expression of Flag-S was detected by Western blot. HEK293 cells were divided into the cells 1, 2, 3 and 4 and transfected with GFP11 plasmid and vector, GFP1-10 plasmid and vector, transfected with GFP11 and pCMV-HA-ACE2 plasmid, GFP1-10 and p3xflag-S plasmid. Cell 1 was co-cultured with cell 2 (control group 1), cell 2 with cell 3 (control group 2), cell 3 with cell 4 (observation group), and cell 1 mixed with cells 2, 3 and 4 (control group 3). Bright-field microscopy and fluorescence microscopy were used to observe cell fusion. RPE cells were divided into control group and overexpression S-protein group. The cell cycle was detected by flow cytometry; the cell proliferation level was detected by Counting Kit 8 (CCK-8); and the S-protein expression level in RPE cells was detected by Western blot. The Student’s t-test was performed for comparison between groups. Results:DNA sequence assay showed that S-protein cDNA was fused with flag-tagged protein. Western blot assay showed that S-protein-related expression was elevated in transfected HEK293 cells compared with untransfected p3xflag-S cells. Large, multinucleated fused cell clusters were visible under bright-field microscopy; multiple nuclear with distinct green fluorescence were visible in the fused cells under fluorescence microscopy. Western blot assay showed elevated S-protein-related expression in transfected p3xflag-S plasmid RPE cells compared to untransfected p3xflag-S plasmid RPE cells. CCK-8 results showed that the proliferative capacity of RPE cells in the S-protein overexpression group was significantly reduced compared with the control group, with statistically significant differences ( t=22.70, 16.75, 23.38; P<0.000 1). The results of flow cytometry showed that the G1 phase cells in the control and overexpression S-protein groups were 41.1 % and 67.0%, respectively; compared with the control group, the G1 phase cells in the overexpression S-protein group were significantly higher, and the difference was statistically significant ( t=4.76, P=0.018). The apoptosis rate was significantly increased in the S-protein overexpression group compared with the control group, and the difference was statistically significant ( t=4.91, P=0.008). Conclusion:Overexpression of the SARS-CoV-2 spike protein reduced the proliferation of human RPE cells.

Objective:To investigate the anti-aging and antioxidant effect of the heat shock transcription factor 1 (HSF1) on human retinal pigment epithelial cells.Methods:Two HSF1-deficient ARPE cells (ARPE/Hsf1 -/-) were constructed by using the clustered regularly interspaced short palindromic repeat and associated protein 9 (CRISPR/Cas9) gene editing system and named H8, H9 konckout cell strains.Experiments were operated on the 3 cell strains: wild-type, H8 and H9 cells.The content of reactive oxygen species in ARPE-19 cell was measured by DHE probe staining combined with flow cytometry technology, and the cell cycle was measured by flow cytometry technology.The cell viability at different time points was measured using cell counting kit-8 (CCK-8).Crystal violet staining assay was used to measure the relative ratio of cell survival.SA-β-gal staining assay was used to detect the ratio of ARPE-19 senescent cells.The expressions of HSP70, HSP27, clusterin (CLU), p53, p21 and interleukin (IL)-1β proteins were measured by Western blot technology.The expressions of p53, p21, IL-6, IL-8, IL-1β and monocyte chemoattractant protein 1 (MCP1) mRNA were measured by quantitative real-time PCR technology.Relative expression of heat shock response protein under different heat shock treatment conditions and HSP90 inhibitor IPI504, relative survival with different concentrations of H 2O 2, relative expression of p21 protein after treatment with or without ROS scavenger N-acetylcysteine (NAC) were compared in each cell strain. Results:Gene sequencing showed that H8 and H9 cell strains successfully carried mutated genes.Western blot experiment results showed that H8 and H9 cell strains did not express HSF1 protein, and HSF1 was successfully knocked out in ARPE-19 cells.Compared with wild-type cell, the expression levels of HSP70, HSP27 and CLU proteins in H8 and H9 cell strains significantly decreased, with statistically significant differences (all at P<0.05), and no significant difference was found in the relative HSP90 protein expression level ( F=0.29, P>0.05).Under different heat shock stimulation and IPI504 induction, the HSP70, HSP27, and CLU protein levels significantly increased in wild-type cells compared with before treatment, and the HSP70, HSP27, and CLU protein levels were significantly lower in H8 and H9 cell strains than in corresponding treated wild-type cells (all at P<0.05).Compared with wild-type cell strains, cell viability significantly decreased in H8 and H9 cell strains at 24, 48, 72, and 96 hours (all at P<0.05).Compared with wild-type cell strains, the percentage of cells in G1 phase was significantly higher and the mRNA and protein levels of the cell cycle inhibitors p53 and p21 significantly increased in H8 and H9 strains, showing statistically significant differences (all at P<0.05), and the ratio of positive cells for SA-β-gal staining significantly increased, showing statistically significant differences (all at P<0.001).The relative expression of aging-related inflammatory factors IL-6, IL-8, IL-1β, and MCP1 mRNA decreased, and the differences were statistically significant (all at P<0.001).In addition, compared with wild-type cell strains, the content of reactive oxygen species (ROS) was higher in H8 and H9 cell strains, and the differences were statistically significant (all at P<0.001).The expression of p21 protein in H8 and H9 cell strains wtih NAC treatment decreased significantly compared with non-NAC treatment cells (both at P<0.05).Compared with wild-type cell strains, H8 and H9 cell viability decreased at 200, 400, 600, and 800 μmol/L H 2O 2 treatment conditions, and the differences were statistically significant (all at P<0.05). Conclusions:Knockdown of HSF1 can downregulate the expression of heat shock proteins, activate the ROS/P53/P21 pathway, induce senescence in RPE cells, and increase the sensitivity of RPE to oxidative stress stimuli.HSF1 may have anti-senescence and anti-oxidant regulatory effects in RPE cells.