BACKGROUND:Neural stem cells (NSCs) hold self-renewal and multi-directional differentiation potential. NSCs differentiation into neurons in high proportion under induction conditions exhibits broad application prospect. OBJECTIVE:To explore the effect of basic fibroblast growth factor (bFGF)-chitosan carrier on the NSCs differentiation into neurons in vitro, and whether the differentiated neurons could form synaptic-like connection with myocytes. METHODS:After purification, the NSCs were co-cultured with chitosan, soluble bFGF or bFGF-chitosan carrier. After 7-day induction, the NSCs differentiation into neurons was observed by immunofluorescence staining of beta tubulin Ⅲ. The NSCs differentiation into cholinergic neurons was observed through double immunofluorescence staining of ChaT and beta tubulin Ⅲ. The synaptic-like connection between the neurons and myocytes was observed by triple staining of beta tubulin Ⅲ and MHC. RESULTS AND CONCLUSION:The percentage of differentiated neurons in the bFGF-chitosan carrier group was 74%, which was significantly higher than that in the other two groups. Additionally, the synaptic-like connection formed between the differentiated neurons and myocytes. To conclude, the bFGF-chitosan carrier promotes the NSCs differentiation into neurons to form synaptic-like connection with the co-cultured myocytes.

Objective:To establish two differential gene expression profiles of qi-deficiency and blood stasis syndrome before or after safflower injection treatment by using gene chip technology;compared and analyzed to ensure the effective genes that are responsible for the therapeutic effects of safflower injection against qi-deficiency and blood stasis syndrome in rats.Furthermore,speculated the effect mechanism of the therapeutic genes.Methods:Fifteen SD rats were randomly divided into 3 groups (n=5):control group,model group,and medication group.Qi-deficiency and blood stasis model was established by subjecting the rats to hunger and fatigue for two weeks.After a week of the modeling,safflower injection (100 mg/kg/d) was administered daily via the tail vein for 7 days in medication group,and the rats in model group were injected with saline of the same volume.Control group received normal feeding.At the end of the experiment,rats were killed and whole blood was collected to evaluate the blood stream change and extract mRNAs in blood samples.Qualified mRNAs were reverse transcribed into cDNA which was then used in gene chip hybridization.The genes regulated by safflower injection were determined by the fluorescence signal and the functional mechanisms of safflower injection were confirmed by further querying genealogy databases and reviewing literatures.Results:After two weeks of the modeling,the whole blood viscosity under various shear rates was significantly increased in the model rats which showed faint,blood stasis and weight loss,indicating that the model is made successfully.The increased whole blood viscosity and qi-deficiency and blood stasis syndrome were obviously reversed by safflower injection treatment.Compared with the control group,252 genes up-regulated while 54 genes down-regulated in model group;compared with the model group,196 genes up-regulated while 32 genes down-regulated.Among these,16 differentially expressed genes were involved in inflammation and immune response.Conclusions:Safflower injection was effective in treating qi deficiency and blood stasis syndrome,which was achieved by regulating inflammation related genes.

Objective:In order to develop an oral live vaccine vector of swine that can stably carry exogenous genes.Methods:Mutant ΔcrpΔcyaΔasdC78-1 was constructed by the method of suicide plasmid pREasd-mediated bacteria homologous recombination on the basis of attenuated Salmonella choleraesuisΔcrpΔcyaC78-1.Complementary plasmid pYA3493 with asd was electrotransformed into the mutant,and thenΔcrpΔcyaΔasdC78-1(pYA3493) host-vector balanced lethal system was constructed.Its biological characteristics were analyzed further.Results:The results of PCR and sequencing showed thatΔcrpΔcyaΔasdC78-1(pYA3493) was constructed suc-cessfully.Biological characteristics showed that the serotype of ΔcrpΔcyaΔasdC78-1(pYA3493) was identical to ΔcyaΔasdC78-1 and vaccine strain C500 and it can stably carry theΔasd gene in vitro;its growth speed was a little slower than ΔcrpΔcyaC78-1 strain,but both of their growth speeds were significantly slower than vaccine strain C500;the biochemical characteristics of ΔcrpΔcyaΔasdC78-1 ( pYA3493 ) were basically the same with ΔcrpΔcyaC78-1 strain.Oral virulence test in mice showed that the virulence ofΔcrpΔcyaΔasdC78-1 ( pYA3493 ) was similar with ΔcrpΔcyaC78-1, but its median lethal dose is 412 times of vaccine strain C500.Conclusion:These results demonstrated that attenuated Salmonella choleraesuisΔcrpΔcyaΔasdC78-1(pYA3493) strain had the potential to be used as an oral live vaccine vector for expressing foreign genes efficiently.

Objective To evaluate the outcome of ST-segment elevation myocardial infarction (STEMI) patients received different reperfusion therapies. Methods The 238 consecutive STEMI patients were enrolled from February 2012 to December 2012. According to the current guideline of PCI and the choice of patients, the patients were divided into the groups of percutaneous coronary intervention (PCI), ifbrinolysis, and conservative medication. The major adverse cardiac events (MACE) was analyzed in a follow up of 6 months. Results (1) The enrolled patients included the 210 patients received PCI (88.2%), 14 patients received fibrinolysis (5.9%) and 14 patients received conservative medication (5.9%).The Median time of D2B was 110minutes.(2) The rate of late stent thrombosis was signiifcant higher in BMS than DES (n=2, 2.8%vs 0, P < 0.05) . (3) The PCI group had a signiifcantly higher incidence of stroke than the ifbrinolysis group and the conservative medication group (1.0%vs 0, P < 0.05;1.0%vs 0, P<0.05). (4) The PCI group had a signiifcantly higher incidence of bleeding compared to the thrombolysis group and the medication group (1.0% vs 0, P < 0.05; 1.0% vs 0%, P < 0.05). Conclusions The majority of STEMI patients received PCI;The D2B time, which was required<90 minutes in guideline of PCI, was found delayed in our study;Compared to ifbrinolysis and conservative medication, PCI showed better clinical outcomes of STEMI patients.

Objective To explore the relationship between pulse blood pressure and cognitive function for the elders in Yuhuan County,Zhejiang Province.Methods Multi -stage cluster method was used to select 1 700 residents aged over 60 years in Yuhuan County,Zhejiang Province.Mini -Mental State Examination (MMSE)was used to screen cognitive impairment.Locally weighted scatter plot smoothing was used to explore the local linear trend between MMSE score and diastolic.The association between pulse pressure or blood pressure and cognitive function was analyzed by multivariate logistic regression analysis.Results The prevalence of cognitive impairments for the elders aged over 60 years in Yuhuan County was 15.74%.There was significant difference between the pulse blood pressure of cognitive impairment group (64.01 ±16.31)mmHg and none -cognitive impairment group (58.97 ±14.67)mmHg.The t value was 4.97.Pulse blood pressure and the elderly MMSE score showed a significant positive correlation.Multivariate logistic regression analysis showed that cognitive function related to diastolic blood pressure (OR =1.010,95% CI:1.001 -1.019,P =0.034)after adjusting several covariate.Conclusion Normal cognitive function was associated with decreased pulse blood pressure among the elders aged over 60 years in Yunhuan County,Zhejiang Province.

Objective To investigate the characteristics of blood routine in 129 patients with COVID-19, and analyze the correlation between blood routine parameter changes and clinical classification. Methods A total of 129 COVID-19 patients were recruited and their blood samples were collected at the beginning and the end of treatment.The neutrophil-to-lymphocyte ratio(NLR), platelet-to-lymphocyte ratio(PLR), lymphocyte-to-monocyte ratio(LMR) and C-reactive protein(CRP) were determined and used to calculate Δ NLR, Δ PLR, Δ LMR.The differences in these parameters were compared between the non-severe group (93 cases) and the severe group (36 cases).In addition, the relationship between the changes in blood routine test result and the prognosis of patients was determined. Results The average age of 129 patients was 46.9±17.4 years old, and the ratio of male to female was 1.2 ∶ 1.Thirty-five (35) cases (27.1%) had leucopenia (< 4×10⁹/L) and 59 cases (45.7%) had lymphopenia (< 1.1×10⁹/L).There were statistically significant differences in age, treatment days, blood routine indexes between these two groups.In all the patients, the differences between before and after treatment were statistically significant in the following: leukocyte number, neutrophil cell percentage, lymphocyte number, lymphocyte percentage, monocyte number, monocyte percentage.The differences in RBC, HGB, CRP, and NLR between the two groups before and after treatment were also statistically significant (P < 0.05). Conclusion The older COVID-19 patients have lower number of lymphocytes, higher NLR and PLR, lower LMR and higher CRP.They have a higher risk of progressing to severe disease.After treatment, there is an increase in the number of granulocytes, especially the number of lymphocytes, while a decrease in CRP and NLR.The change of lymphocyte count, NLR and CRP levels can predict the risk of severe COVID-19 and evaluate the therapeutic effect.

OBJECTIVETo investigate, at the DNA level, the polymorphism of HLA-A, -B, -Cw genes in the Chinese of Han ethnicity in Shenyang.

METHODSHybridization with polymerase chain reaction-sequence specific oligonucleotide probes (PCR-SSOP) was used to determine HLA-A, -B and -Cw genotypes of 108 unrelated healthy individuals from a Chinese Han population. These Hans were born and living in the Shenyang area.

RESULTSThe numbers of alleles identified were 21 for HLA-A, 43 for HLA-B, and 23 for HLA-Cw. All the allele frequency distributions were consistent with the Hardy-Weinberg equilibrium.

CONCLUSIONUsing molecular method, the present authors have analyzed the characteristic of HLA I distribution in a group of indigenous Hans in Shenyang and thus have provided more accurate gene data for use in related researches.

Adolescent ; Adult ; Alleles ; China ; DNA ; analysis ; genetics ; Female ; Gene Frequency ; Genotype ; HLA-A Antigens ; genetics ; HLA-B Antigens ; genetics ; HLA-C Antigens ; genetics ; Humans ; Linkage Disequilibrium ; Male ; Middle Aged ; Oligonucleotide Probes ; Polymerase Chain Reaction ; methods ; Young Adult

Objective:To study the medication rules of treating cervical spondylosis by National TCM master Liu Bailing based on data mining and network pharmacology, and explore the potential action mechanism of its core compounds.Methods:By collecting the prescriptions of National TCM master Liu Bailing treating cervical spondylosis in the past 8 years, this paper analyses the frequency, nature, flavor, meridian, hierarchical clustering and association rules of those prescriptions by RStudio to obtain the core prescription. Then, the effective components of the core prescription were collected by using TCMSP, and the network of "medicine-component-target" was constructed by using Cytoscape 3.8.0; by searching for databases like GEO, DisGeNET, TTD HPO and Genecards were retrieved to obtain the target data set of cervical spondylosis; by using STRING 11.0 platform to construct protein interaction network; by using DAVID platform to cary out gene ontology (GO) and KEGG pathway enrichment analysis; by using Auto Dock software for molecular docking.Results:In the 844 prescriptions, there are 199 Chinese medicines and the properties are mainly warm, plain and cold; the flavors were mainly sweet, pungent and bitter; mainly belong to the liver, spleen, and kidney meridians. The Association Rule shows that the core compound is made up of Salvia miltiorrhiza, Gastrodia elata, Rhizoma corydalis, Alisma rhizoma, centipede, Astragalus membranaceus and Rhizome of Pueraria. Besides, 140 effective constituents and 247 targets of the core prescription were screened, and the main constituents were quercetin, kamanol, luteolin, tanshinone ⅡA, β-sitosterol, etc. 13 core targets among the core prescription treating cervical spondylosis were obtained, which were enriched into 30 pathways including toll-like receptor signaling pathway, TNF signaling pathway and HIF-1 signaling pathway. Conclusion:National TCM master Liu Bailing treatment of cervical spondylosis mainly focuses on expelling wind and relieving pain, dredging meridians and soothing tendons, and the mechanism of action of the core prescription may focus on inhibiting inflammatory response and relieving oxidative stress, providing guidance and reference for the clinical treatment of cervical spondylosis.

Objective:To explore the impact of serum carbamoyl phosphate synthase 1 (CPS1) level on prognosis of patients with hepatitis E-related acute liver failure (HEV-ALF).Methods:This retrospective analysis included 100 HEV-ALF patients, 100 patients with acute hepatitis E (AHE) and 100 healthy controls who admitted or underwent health checkup from December 2018 to June 2019 in six hospitals, including the First Affiliated Hospital, Zhejiang University School of Medicine. HEV-ALF patients were divided into non-survial ( n=21) and survival ( n=79) subgroups according to results of 30-day follow-up results. HEV-ALF patients were also divided into the high ( n=50) and low ( n=50) serum CPS1 level groups. HEV-ALF patients were further divided into the improvement ( n=55), fluctuation ( n=32) and deterioration ( n=13) subgroups. The general clinical data from all participants were collected. Serum CPS1 levels were detected by enzyme linked immunosorbent assay. The survival time in the high and low serum CPS1 level groups were presented in the Kaplan-Meier curve. The correlation between serum CPS1 level and HEV-ALF related conventional parameters was also analyzed by linear regression. The efficacy of serum CPS1 level on predicting the 30-day mortality of HEV-ALF patients was estimated by the receiver operating characteristic curve and area under curve (AUC). Results:Serum CPS1 level was significantly higher in HEV-ALF patients than in AHE patients [958.59 (665.52, 1 105.83) pg/ml vs 549.38 (495.02, 649.08) pg/ml, P<0.001], and serum CPS1 level was significantly higher in AHE patients than in healthy controls [549.38 (495.02, 649.08) pg/ml vs 469.89 (373.32, 564.53) pg/ml, P<0.001]. The level of serum CPS1 was significantly lower in the HEV-ALF survival group than in the HEV-ALF non-survival group [922.6 (652.7, 1, 042.3) pg/ml vs 1 252.8 (933.3, 1 555.8) pg/ml, P<0.001]. In addition, the survival time was shorter in the high serum CPS1 level group than in the low serum CPS1 level group [24.59 (22.11, 27.06) d vs 28.16 (26.25, 30.07) d, P=0.045]. Serum CPS1 levels were increased in the fluctuation and deterioration groups [Fluctuation: 1 328.3 (1 184.3, 1 964.0) pg/ml vs 1 245.7 (1 102.0, 1 937.6) pg/ml, P<0.01; Deterioration: 1 483.6 (1 275.9, 1 656.8) pg/ml vs 1 332.2 (1 197.4, 1 509.8) pg/ml, P<0.01], while decreased in the improvement group [810.3 (599.7, 904.5) pg/ml vs 922.6 (679.5, 1 039.6) pg/ml, P<0.01] over time. Besides, a linear positive correlation was found between serum CPS1 level and alanine aminotransferase (ALT) and total bilirubin (TBIL) (ALT: r=0.339, P<0.001; TBIL: r=0.304, P=0.002). The AUC of serum CPS1 level to predict the 30-day mortality of HEV-ALF patients was 0.803 (95% CI 0.666-0.941), the sensitivity and specificity were 66.67% and 97.47%, respectively. Conclusion:Serum CPS1 level was significantly increased in HEV-ALF patients, and closely related to the prognosis of patients with HEV-ALF.

OBJECTIVETo investigate the levels of blood pressure and serum lipids, and examine the relationship between hypertension and hyperlipidemia in Hei Yi Zhuang Chinese living in Guangxi.

METHODSA total of 1056 people of Hei Yi Zhuang ethnicity were studied. Blood pressure, body height, body weight, and serum levels of lipids and apolipoprotein were measured. The data were compared with those in 925 people of Han ethnicity, who live in the same region.

RESULTSSystolic blood pressure and pulse pressure were significantly higher in Hei Yi Zhuang than Han Chinese (P < 0.001). The prevalence of isolated systolic hypertension and hypertension was also significantly higher in Hei Yi Zhuang than Han Chinese (P < 0.001). Serum concentrations of total cholesterol, triglycerides, low-density lipoprotein cholesterol, and apolipoprotein (Apo) B, and the prevalence of hypercholesterolemia and hyperlipidemia were significantly lower in Hei Yi Zhuang than Han Chinese (P < 0.05). Serum concentrations of high-density lipoprotein cholesterol and the Apo A1 to Apo B ratio were significantly higher in Hei Yi Zhuang than Han Chinese (P < 0.001). The prevalence of hypertension in Hei Yi Zhuang Chinese was positively associated with triglycerides (r = 0.425, P < 0.05), whereas the prevalence of hypertension in Han Chinese was positively correlated with total cholesterol (r = 0.623, P < 0.001).

CONCLUSIONThe present study revealed a significant difference in blood pressure and serum lipids between Hei Yi Zhuang and Han ethnic groups, and an association between hypertension and hyperlipidemia.

Adult ; Aged ; Asian Continental Ancestry Group ; ethnology ; Blood Pressure ; China ; epidemiology ; Female ; Humans ; Hyperlipidemias ; epidemiology ; ethnology ; Hypertension ; epidemiology ; ethnology ; Lipids ; blood ; Male ; Middle Aged ; Prevalence ; Sampling Studies ; Young Adult