Objective To observe whether the increase of oxidative stress in PC12 cells could influence the levels of protein carbonyls and nitrotyrosine and alter the cytoskeleton and cell morphology under clinostat condition,and whether the increase of content of nitrate/nitrite in cell culture medium could influence the cell proliferation and differentiation.Method Cell morphology,carbonylated actin and nitrotyrosinated tubulin,and mRNA and protein express of nNOS and iNOS were observed and determined with immunofluorescence and RT-PCR technology in clinostat rotated and control static groups.At the same time,cell density was measured and cell cycles were detected with flow cytometry.The relationship between all these changes and NOS were also analyzed.Result The levels of carbonylated and nitrotyrosinated cytoskeleton protein were altered,no obvious changes in cell morphology but neurite outgrowth after on a clinostat rotation.Cell density also increased significantly,DNA synthesis in cell cycles was shortened.Conclusion All of these results indicate that simulated weightlessness do not alter cell morphology and is beneficial to the growth of PC12 cells.The mechanism involved may be associated with the increase of NOS activity.

Objective To establish an HPLC-ELSD method for the determination of pinitol in Fufang Kushen Injection. Methods Chromatographic condition was as follows:Cosmosil Sugar-D column (250 mm×4.6 mm, 5 μm), the mobile phase consisting of acetonitrile-water in gradient mode, the temperature of the drift tube of 77 ℃, and the gas flow rate of 2.0 L/min. Results The standard curve of pinitol was rectilinear within the range of 0.5-10.0 μg, r=0.999 6 (n=5). The average recovery was 97.30%(RSD=1.29%, n=5). Conclusion The method is simple and rapid, the result is accurate, reliable and reproducible.

Objective:To analyze the clinical phenotype of adult patients with epidemic encephalitis B (encephalitis B) in Ningxia Hui Autonomous Region, and to explore the influence of related factors of the development of encephalitis B.Methods:The medical records of confirmed patients with encephalitis B admitted to the General Hospital of Ningxia Medical University from August to November 2018 were collected, and the general data of patients and the results of laboratory indexes such as blood routine examination and cerebrospinal fluid routine examination were analyzed. Logistic regression analysis and survival curve were used to evaluate the risk factors of the development of encephalitis B.Results:Totally 97 patients with encephalitis B were included, 32 of them died, with a case fatality rate of 32.99%. There were 63 males and 34 females, and the age of onset was (59.13 ± 14.70) years old. There were statistically significant differences in case distribution rate between different sexes and ages (χ 2 = 97.00, 291.00, P < 0.001). The most common clinical type was extremely severe (43 cases), followed by mild (27 cases), severe (15 cases) and ordinary (12 cases). The results of laboratory tests showed that the number of neutrophils, lymphocytes and monocytes in the blood of patients increased; and the white blood cells number in cerebrospinal fluid increased significantly, while neutrophils ratio increased slightly. There were significant differences in cerebrospinal fluid glucose level and neutrophil ratio among patients with different clinical types of encephalitis B ( H = 4.21, 2.74, P < 0.05). There were statistically significant differences in death, hypertension, cerebrovascular diseases, and pulmonary infection among patients with different clinical types of encephalitis B (χ 2 = 34.22, 16.97, 9.91, 15.59, P < 0.05). Logistic regression analysis showed that hypertension [ OR (95% CI) = 5.544 (1.450-21.191)] and pulmonary infection [ OR (95% CI) = 6.490 (1.887-22.325)] were risk factors for the development of encephalitis B patients ( P = 0.012, 0.003). Pulmonary infection was the influencing factor for the death of encephalitis B patients (χ 2 = 18.88, P < 0.001). The survival curve showed that the survival status of encephalitis B patients with cerebrovascular disease and pulmonary infection was significantly worse than that of patients without comorbidity or complications (χ 2 = 6.45, 20.33 , P < 0.05). Conclusions:The majority of encephalitis B patients in this outbreak are the elderly people, and the patient's nervous system has inflammatory reaction. Complicated pulmonary infection is an important factor for the aggravation and death of encephalitis B patients.

Objective To study the genotyping distribution of the Yersinia pestis(Y.pestis)strains by characterizing the diversity of the insertion sequence IS100 within the Y.pestis genome.Methods Derived fromthe known sequence of oriental strain CO92,5 pairs of locus-specific primers originating from both sides of the adjacent region of IS100 copies were designed,and two other complementary primers inside the IS100 sequence were designed to correspond with the outer primers.Then,91 Y.pestis strains and l pseudotubebculosis strain were tested by the specific PCR method using the primers described above and the PCR products were conformed by the sequence analysis,then further analysis WaS performed after the IS100 status was marked on the map of the plague focus type of china.Results The 91 Y.pestis strains had different IS100 status in their genome on tested loci.some possessed IS100 insertion,some didn't,and others changed their genome constitution.The IS100 possession on the 5 loci also suggested a distribution of regionality.Conclusion The analysis of some IS100 insertion element loci reveals that the IS100 genotyping distribution is consistent with the plague focus of type of China.And IS100genotyping pattern of the Y.pestis stains well reflects its genome constitution and the high flowability in its natural evolution.

Objective:To compare the clinical efficacy and safety of conventional transcatheter arterial chemoembolization (TACE) with drug-eluting bead transcatheter arterial chemoembolization (DEB-TACE) in treatment of patients with unresectable hepatocellular carcinoma.Methods:The data of patients with unresectable hepatocellular carcinoma who underwent hepatic artery chemoembolization at General Hospital of Ningxia Medical University from July 2019 to April 2020 were retrospectively analyzed. Of 282 patients who were enrolled, there were 233 males and 49 females, aged (55.9±10.0) years. The groups were divided into the conventional TACE group ( n=179) and the DEB-TACE group ( n=103) based on the treatments. The efficacy of the two groups was compared according to the modified response evaluation criteria in solid tumors. Postoperative adverse effects and liver function between the two groups were compared. Results:The differences in comparing the preoperative and postoperative liver function indexes between the two groups were not statistically significant. Patients who died and were lost to follow-up at 6 months after surgery were excluded and 240 patients were excluded in the efficacy analysis, with 148 patients in the conventional TACE group and 92 patients in the DEB-TACE group. At 6 months after treatment in the conventional TACE group, there were 64 patients (43.2%) with complete remission, 18 patients (12.2%) with partial remission, 27 patients (18.2%) with stable disease, and 39 patients (26.4%) with disease progression. In the DEB-TACE group, the corresponding figures were 38 patients (41.3%), 17 patients (18.5%), 26 patients (28.3%), and 11 patients (12.0%), respectively. The efficacy of DEB-TACE was better than conventional TACE with statistically significant differences between the 2 groups (χ 2=8.96, P=0.030). The incidence of postoperative embolic syndrome was 53.1% (95/179) in the conventional TACE group, which was significantly higher than the 34.0% (35/103) in the DEB-TACE group (χ 2=7.34, P=0.007). Conclusion:The efficacy and safety of DEB-TACE for unresectable hepatocellular carcinoma were superior to those of the conventional TACE group.

Objective To investigate the learning curve of contrast-enhanced ultrasonography ( CEUS) in sentinel lymph node( SLN ) of breast cancer and provide a theoretical basis for leaners to learn SLN CEUS . Methods The multi-center study of SLN CEUS in breast cancer" was planned by Sichuan Cancer Hospital . According to the uniform inclusion and exclusion criteria , 511 patients with complete clinical data and follow-up results from 9 hospitals in Multi-center were included in this study . According to the inspection time ,the patients were divided into 3 groups named as group A ( 170 patients) ,group B ( 170 patients) and group C ( 171 patients ) ,respectively . The basic clinical data ,ultrasound imaging data , intraoperative and postoperative pathological findings of all patients were recorded . With the accumulation of cases examined ,analysis was performed to find the learning curve of the SLN CEUS examination time , SLN CEUS detection rate ,SLN CEUS surface marking accuracy rate and SLN CEUS diagnosis rate ,the learning curve was analyzed . Results ① There was no statistical significant difference in patients ages , tumors sizes ,tumors locations ,SLNs numbers and LCs numbers among the three groups( all P > 0 .05) . ②As the number of cases examined increases ,the examination time was reduced gradually ,but SLN detection rate ,surface marking accuracy and SLN diagnostic coincidence rate were increased gradually( F = 151 .75 , 1 .96 ,7 .49 ,5 .50 ; P = 0 .000 ,0 .143 ,0 .001 ,0 .005 ) . Conclusions The skill of the doctor is improved gradually when learning SLN CEUS . With the number of the cases increase ,the operating time of SLN CEUS is shorted ,and the SLN detection rate ,surface marking accuracy and SLN diagnostic coincidence rate of SLN-CEUS are gradually increased . It has an important clinical significance for beginners to learn the SLN CEUS technology .

ObjectiveTo investigate the regulatory effect of miRNA-933 on the apoptosis and proliferation of human hepatic stellate cell line LX-2 and its mechanism. MethodsFirstly， with human liver tissue for research， gene microarray technology was used to detect the differentially expressed genes in liver tissue between liver cirrhosis/chronic hepatitis B tissue and normal liver tissue， among which the significantly differentially expressed miRNAs were identified， and thus miRNA-933 was determined as the research object. Then， with the human hepatic stellate cell line LX-2 for research， miRNA-933 mimic and inhibitor （miRNA-933 siRNA） were used to construct the LX-2 models of overexpression and knockdown， and the cells transfected with mimic-NC （overexpression） or siRNA-NC （knockdown） were established as the negative control group. Quantitative real-time PCR and Western blot were used to measure the expression levels of miRNA-933 and activation biomarkers； techniques such as cell proliferation assay and flow cytometry were used to investigate the effect and mechanism of miRNA-933 on cell apoptosis， proliferation， and activation. The independent-samples t test was used for comparison of continuous data between two groups； a one-way analysis of variance was used for comparison between multiple groups， and Bonferroni correction was also performed. ResultsA total of 18 significantly differentially expressed miRNAs were obtained based on the results of gene microarray， among which miRNA-933 was significantly downregulated （P<0.05）. After LX-2 cells were transfected with miRNA-933 mimic or siRNA， compared with the negative control group， miRNA-933 siRNA significantly downregulated the expression of miRNA-933 （P=0.000 7）， while miRNA-933 mimic significantly upregulated the expression of miRNA-933 （P=0.000 3）. Western blot and quantitative real-time PCR showed that miRNA-933 siRNA significantly upregulated the expression of collagen I and α-SMA （P<0.001）， while miRNA-933 mimic significantly inhibited the expression of collagen I and α-SMA （P<0.05）. Flow cytometry showed that compared with the negative control group， miRNA-933 siRNA significantly downregulated the apoptosis rate of LX-2 cells （P=0.031 9）， and miRNA-933 mimic significantly upregulated the apoptosis rate of LX-2 cells （P=0.005 5）. Western blot showed that compared with the negative control group， miRNA-933 siRNA could inhibit the expression of Caspase-3 （P=0.006 7） and poly（ADP-ribose） polymerase-1 （PARP-1） （P=0.003 0） and upregulate the expression of B-cell lymphoma-2 （Bcl-2） in LX-2 cells （P=0.002 0）， while miRNA-933 mimic could significantly upregulate the expression of Caspase-3 （P=0.011 8） and PARP-1 （P=0.049 5） and downregulated the expression of Bcl-2 （P=0.002 1）. Cell proliferation assay showed that compared with the negative control group， miRNA-933 siRNA could promote the proliferation of LX-2 cells （P=0.011 5）， while on the contrary， miRNA-933 mimic could inhibit the proliferation of LX-2 cells （P=0.001 2）. Western blot and quantitative real-time PCR showed that miRNA-933 siRNA significantly inhibited the expression of Kruppel-like factor 6 （KLF6） and downregulated the expression of activating transcription factor 4 （ATF4）， activating transcription factor 3 （ATF3）， and C/EBP homologous protein （CHOP）， while miRNA-933 mimic promoted the expression of the above proteins （all P<0.05）. ConclusionThis study shows that miRNA-933 may promote cell apoptosis and inhibit cell activation and proliferation by promoting the activation of the KLF6/ATF4/ATF3/CHOP/Bcl-2 signal axis in LX-2 cells.

AIM: To explore the mechanism of action of alkaloid components of compound kushen Injection (CKI) in the treatment of lung cancer based on serum metabolomics, network pharmacology, and molecular docking techniques. METHODS: A lung cancer model was established in C57 mice by inoculation of Lewis mouse lung cancer tumor strain. Thirty male mice were randomly divided into normal group, model group and CKI group. The drug was administered by tail vein injection once daily for 17 consecutive days. Mouse serum was examined by ultrahigh performance liquid chromatography tandem mass spectrometry (LC-MS) metabolomics, and several multivariate statistical analyses including principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA), combined with databases such as the human metabolic database (HMDB) and related literature to identify and identify differential metabolites, the relevant metabolic pathways were searched for by the metaboanalyst online tool. Using network pharmacology, construct the“component-target-disease”network of CKI in the treatment of lung cancer. Molecular docking method was used to verify the interaction between potential active ingredients and core targets. Serum metabolomics was jointly analyzed with network pharmacology to construct a“metabolite-germinal-enzyme-gene” network. RESULTS: Through metabolomics technology, 16 differential metabolites associated with lung cancer were screened from serum, and CKI addback these differential metabolite levels compared with the model group. Metabolic pathways mainly involve retinol metabolism, tryptophan metabolism, glycerophospholipid metabolism and other metabolic pathways. Network pharmacology analysis indicated that CKI treatment of lung cancer mainly targets STAT3, MAPK3, and MAPK1, which are closely related to proteoglycans, cellular senescence, and HIF − 1 signaling pathways in cancer. CONCLUSION: This article explains the mechanism of CKI in treating lung cancer from the perspective of metabonomics and network pharmacology, and provides basis for further study of CKI.

Objective: To investigate the diagnostic value of serum α-enolase (ENO1) in the primary hepatocellular carcinoma. Methods: From May 2012 to March 2017, 163 cases with liver diseases who met the inclusion and exclusion criteria were admitted to the Infectious Diseases Department of the General Hospital of Ningxia Medical University. Among them, 28 cases were of chronic hepatitis B (CHB), 31 cases with liver cirrhosis (LC), 104 cases with hepatocellular carcinoma (HCC), and 18 healthy volunteers (NC). Patient data and serum samples were collected and liver disease related indicators were measured to detect ENO1 levels with enzyme-linked immunosorbent assay (ELISA). The measured indicators were expressed in median. Mann-Whitney U nonparametric test was used to analyze the differences between the data. A Spearman’s correlation analysis was used for bivariate correlation analysis. The sensitivity and specificity of ENO1 and alpha-fetoprotein in the diagnosis of liver cancer were analyzed by ROC curve. Results: Serum level of ENO1 in CHB group, LC group and HCC group was significantly higher than normal group. Serum level of ENO1 in HCC group was higher than CHB group (P = 0.001) and LC group (P < 0.01). Area under the curve (AUC) for serum ENO1 and alpha-fetoprotein were 0.782 (cut-off value 75.96, P = 0.000 1) and 0.800 (cut-off value 27.02, P = 0.000 1), respectively. There was a positive correlation between ENO1 and AFP (P = 0.001). The combined detection had significantly improved the detection efficiency (AUC = 0.835). Serum ENO1 was statistically significant (P < 0.05) in HCC tumor size (AUC = 0.663), tumor metastasis (AUC = 0.681), TNM stage (AUC = 0.710, stage I vs. II), and Edmondson grade (AUC = 0.685) (P < 0.05) and the elevated levels of ENO1 had significantly reduced (P < 0.05) the survival time. Conclusion ENO1 can be a new candidate marker for the diagnosis of early stage HCC and its progression.

Background: and Purpose Recent studies suggested an increased incidence of cerebral venous thrombosis (CVT) during the coronavirus disease 2019 (COVID-19) pandemic. We evaluated the volume of CVT hospitalization and in-hospital mortality during the 1st year of the COVID-19 pandemic compared to the preceding year. Methods: We conducted a cross-sectional retrospective study of 171 stroke centers from 49 countries. We recorded COVID-19 admission volumes, CVT hospitalization, and CVT in-hospital mortality from January 1, 2019, to May 31, 2021. CVT diagnoses were identified by International Classification of Disease-10 (ICD-10) codes or stroke databases. We additionally sought to compare the same metrics in the first 5 months of 2021 compared to the corresponding months in 2019 and 2020 (ClinicalTrials.gov Identifier: NCT04934020). Results: There were 2,313 CVT admissions across the 1-year pre-pandemic (2019) and pandemic year (2020); no differences in CVT volume or CVT mortality were observed. During the first 5 months of 2021, there was an increase in CVT volumes compared to 2019 (27.5%; 95% confidence interval [CI], 24.2 to 32.0; P<0.0001) and 2020 (41.4%; 95% CI, 37.0 to 46.0; P<0.0001). A COVID-19 diagnosis was present in 7.6% (132/1,738) of CVT hospitalizations. CVT was present in 0.04% (103/292,080) of COVID-19 hospitalizations. During the first pandemic year, CVT mortality was higher in patients who were COVID positive compared to COVID negative patients (8/53 [15.0%] vs. 41/910 [4.5%], P=0.004). There was an increase in CVT mortality during the first 5 months of pandemic years 2020 and 2021 compared to the first 5 months of the pre-pandemic year 2019 (2019 vs. 2020: 2.26% vs. 4.74%, P=0.05; 2019 vs. 2021: 2.26% vs. 4.99%, P=0.03). In the first 5 months of 2021, there were 26 cases of vaccine-induced immune thrombotic thrombocytopenia (VITT), resulting in six deaths. Conclusions During the 1st year of the COVID-19 pandemic, CVT hospitalization volume and CVT in-hospital mortality did not change compared to the prior year. COVID-19 diagnosis was associated with higher CVT in-hospital mortality. During the first 5 months of 2021, there was an increase in CVT hospitalization volume and increase in CVT-related mortality, partially attributable to VITT.