The current official quality control approaches meet the challenges from the complexity of herbal medicines.In fact,any herbal medicines containing numerous unknown components,its curative effect usually depends on the whole of herbal medicines,so it is impossible and unnecessary to qualitatively and quantitatively study every component.By investigating the limitations of current quality control approaches for herbal medicines and the difference and similarity in the chemical substantial style as well as quality control pattern of herbal medicines,a new quality control approach for Chinese herbal medicines should be explored and designed.The combination approach of chemical analysis with bioassay is promising to be developed and employed in order to ensure the safety and efficacy of Chinese herbal medicines.

To explore the new pattern of Chinese medicine solid preparations (CMSP) in vitro dissolution, a method testing the bio-thermal activity in combination with UPLC was used. Microcalorimetry was used to obtain the characteristic metabolic growth power-time curves and a series of biothermodynamic parameters of the inhibition of Staphylococcus aureus by Yinhuang tablet dissolving solutions at the pH 6.8 (phosphate buffer) dissolution medium at different times. From these results, the cumulative dissolution of Yinhuang tablet based on bio-thermal activity was obtained. The dissolution rates of two components of chlorogenic acid and baicalin were determined by UPLC method. Then f2 similar factor method was used to evaluate the relevance of these two methods. The result showed that f2 values all were more than 50, indicating that there is a good correlation between the two methods of measuring the dissolution rate. It is feasible to determine CMSP in vitro dissolution by using bio-thermal activity, and to provide new evaluation methods for controlling the quality of CMSP.

To compare the microcalorimetric fingerprint profiles of Lonicerae japonicae Flos (Lj.F) and Lonicerae Flos (L.F), microcalormietry was applied to find the heat change regularity of Bacillus shigae (B. shigae) metabolism affected by Lj.F and L.F (we choose Lonicera macranthoides Hand.-Mazz in this paper) with different concentrations. The thermogenic curves and thermodynamics parameters were investigated as evaluation index, and then the date of experiment was studied by similarity analysis. All the results indicated that the Lj.F and Lonicera macranthoides Hand.-Mazz (L.m.H-M) significantly impacted the microbial growth and had good similarity in its inhibitory activities. The combination approach of chemical analysis with bioassay was developed and employed to ensure the safety and efficacy of Chinese herbal medicines.

Objective To investigate the effects of levodopa/benserazide-loaded poly-lactide-coglycolide (PLGA) microspheres on motor deficits and levodopa-induced dyskinesia in a rat model of Parkinson' s disease (PD) and to explore the mechanisms underlying this effects.Methods The content of levodopa/benserazide released from the microspheres was determined by high-performance liquid chromatography.The rat model of PD was induced by 6-OHDA injections.Then the valid PD rats were treated with levodopa ( 12 mg/kg,s.c.)/benserazide ( 15 mg/kg,s.c.) or microspheres.Forepaw adjusting steps were measured on 1,4,7,10 and 14 days after treatment.After 2 weeks of treatment,the abnormal involuntary movements (AIM) were measured.Phosphorylated dopamine,cAMP-regulated phosphoprotein of 32 kDa (DARPP-32) at threonine 34 levels were determined by immunohistochemistry and Western blot respectively.In addition,the levels of △FosB were measured by Western blot.Results In vivo release test showed that 76.2％ of levodopa and 83.6％ benserazide were released from the microspheres on day 7.Forepaw adjusting steps showed that the scores of forepaw adjusting steps in microspheres-treated PD rats were ( 5.8 ± 1.6 ) and ( 5.2 ± 1.5 ) respectively on 10 and 14 days of treatment,which were increased compared to levodopa-treaded PD rats (2.4 ± 1.1 and 1.2 ± 0.5 ; t =4.12,5.43,all P ＜0.01 ).The AIM scores of microspheres-treated rats ( 16.0 ±2.1 ) were decreased significantly compared to levodopa-treated rats ( 26.0 ± 3.2 ) on day 14 ( t =6.59,P ＜ 0.01 ).Immunohistochemistry indicated that the phosphorylated levels of DARPP-32 in microspheres-treated rats ( (3.7 ± 1.3 ) × 104 )were decreased significantly compared to levodopa-treated rats ( (7.9 ± 2.2) × 104 ; t =2.95,P ＜ 0.05 ).In addition,Western blot showed that the levels of phosphorylated DARPP-32 and △FosB were 119.4％ ± 11.3％ and 149.3％ ± 12.3％ respectively,which were decreased significantly compared to levodopatreated rats ( 184.8％ ± 13.7％ and 300.4％ ± 14.2％ ; t =4.12,2.91,all P ＜ 0.05 ).Conclusions Microspheres can be used to improve the motor deficits and reduce the expression of dyskinesia in PD rats.This may be due to the continuous release of levodopa/beserazide from the microspheres,which leads to continuous stimulation of PD rats and reduces the levels of phosphorylated DARPP-32 and △FosB in the striata of these rats.

Objective To observe the biological characteristics of db/db mice, and to provide the basis for application of db/db mice in experimental research.Methods Spontaneous type 2 diabetes BKS.Cg-Dock7m +/+ Leprdb/JNju mice and wild type mice of the same age were used in this study.Their fasting blood glucose was determined at 8,12,16, 20 and 24 weeks of age, the body weight was recorded at 10, 12,16, 20 and 24 weeks of age, the levels of serum insulin, total cholesterol and triglyceride were measured, the organ weight and liver coefficient were determined, and the liver and pancreas were taken for pathological examination at 24 weeks of age.Results The db/db mice maintained a high level of fasting blood glucose and body weight.Levels of serum insulin, total cholesterol, and triglyceride were significantly higher than the wild type mice.Compared with wild type mice, liver weight and kidney weight were also significantly increased.Obvious pathological changes of liver and pancreas were observed in 24-week old db/db mice.Conclusions db/db mouse has obvious characteristics of type 2 diabetes, such as hyperglycemia, hyperlipidemia, and hyperinsulinemia, can maintain stable levels of high blood glucose,and is an ideal animal model for experimental study of type 2 diabetes mellitus.

[ ABSTRACT] AIM:To investigate the different dose of perindopril on cardiac function in the rabbits with ische-mic cardiac dysfunction .METHODS:Male rabbits weighing 2.5~3.0 kg ( n=30) were randomly divided into 3 groups (n=10):high dose perindopril group (HD group), low dose perindopril group (LD group) and cardiac dysfunction group (CD group).The Left anterior descending coronary artery of the rabbits was ligatured for model preparation .In HD group, the rabbits were treated with perindopril split normal saline solution (1 g/L)2 mL· kg-1 · d-1 .In LD group, the rabbits were treated with perindopril split normal saline solution (0.33 g/L)2 mL· kg -1 · d-1.In CD group, the rabbits were treated with normal saline solution 2 mL· kg-1 · d-1 .Four weeks after treatment , the cardiac function was measured via echocardiography , the mRNA expression of angiotensin-converting enzyme 2 ( ACE2 ) and angiotensin type 2 receptor (AT2R) was analyzed by real-time PCR, serum angiotensin (Ang)-(1-9) and Ang-(1-7) levels were detected by ELISA. RESULTS:Compared with CD group , the cardiac function of the 2 groups treated with perindopril was significantly im-proved (P<0.01), and more improvement in HD group was observed than LD group (P<0.05).The serum angiotensin ( Ang)-(1-9) and Ang-(1-7) level and the mRNA expression of ACE 2 and AT2R in the 2 groups treated with perindopril were significantly improved (P<0.01).Compared with LD group, the mRNA expression of ACE2 and AT2R and the ser-um levels of Ang-(1-9) in HD group were significant improved (P<0.05), while no difference of serum Ang-(1-7) level was observed.Correlation analysis revealed that the improvement of the cardiac function was associated with serum Ang -(1-9) level, mRNA expression of ACE2 and AT2R (P<0.01), but has no significant correlation with serum Ang-(1-7) lev-el.CONCLUSION:High dose of perindopril may improve more cardiac function in ischemic cardiac dysfunction model in rabbits.The mechanism may relate to increasing serum Ang-(1-7) level to activate AT2R.

Objective:In this study, the clinical data of biliary atresia(BA) and infant intrahepatic cholestasis(IHC) was reviewed, and the utility of gamma-glutamyl transpeptidase(GGT) and liver Young′s modulus in the differential diagnosis of BA and IHC in infants was discussed.Methods:Based on the clinical data of 120 infants with cholestasis treated in the Children′s Hospital Affiliated to Xi′an Jiaotong University, from September 2017 to December 2019, the infants were divided into two groups according to the results of intraoperative cholangiography and follow-up: BA group( n=50); IHC group( n=70). The age, clinical manifestations, laboratory examination results, gallbladder contraction rate, hepatobiliary scintigraphy, liver Young′s modulus, and medical treatment effects were compared between the two groups.The utility of GGT and liver Young′s modulus in the differential diagnosis of BA and IHC was analyzed. Results:The age, alanine aminotransferase(ALT), aspartate aminotransferase(AST), total bile acid, fasting blood glucose, blood ammonia and splenomegaly between the two groups were compared and the results showed no statistical significance( P>0.05). In contrast, there were statistically significant differences( P<0.001) in stool color, liver size, total bilirubin(TB), direct bilirubin(DB), GGT, liver Young′s modulus, positive hepatobiliary scintigraphy, gallbladder contraction rate at 1 hour after meal, and medical treatment effect between the two groups.TB, DB, GGT, liver Young′s modulus and GGT combined with liver Young′s modulus were analyzed using ROC curves, and the area under the curve(AUC) were 0.820, 0.809, 0.906, 0.876 and 0.926, respectively.When GGT exceeded the cut-off value of 198.85 U/L, the sensitivity, specificity, positive predictive value, negative predictive value and accuracy of BA were 82.0%, 84.3%, 78.8%, 86.8% and 83.3%, respectively.When liver Young′s modulus exceeded the cut-off value of 8.6 kPa, the sensitivity, specificity, positive predictive value, negative predictive value and accuracy of BA were 88.0%, 80.0%, 75.9%, 90.3% and 83.3%, respectively.The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of GGT combined with liver Young′s modulus in diagnosing BA were 98.0%, 68.6%, 69.0%, 98.0% and 80.8%, respectively.Multivariate logistic regression analysis found that DB>115.55 μmol/L, GGT>198.85 U/L, and liver Young′s modulus>8.6 kPa were risk factors for BA( OR=9.510, P=0.001; OR=24.634, P<0.001; OR=21.469, P<0.001). Conclusion:GGT and liver Young′s modulus are useful in the differential diagnosis of BA and IHC.If GGT and liver Young′s modulu sexceed the threshold values of 198.85 U/L and 8.6 kPa respectively, it can effectively indicate that the child is BA.

Objective:To investigate the association of single nucleotide polymorphisms(SNPs)in the vitamin D receptor(VDR)gene with influenza susceptibility and severity of disease in children.Methods:Peripheral venous blood was collected from 172 children with influenza A (study group) and 88 healthy children (healthy control group) admitted to Xi ′an Children′s Hospital and Xi ′an Central Hospital from February 2019 to February 2021.Serum 25-hydroxyvitamin D(25-OH-D) level was detected by using 25-OH-D kit.The study group was divided into three groups according to clinical syndrome: mild group, severe group, and critical group.Four candidate loci in the VDR gene(ApaI, TaqI, FokI, and BSMI)were selected, and polymorphisms in the VDR gene of each group were determined by polymerase chain reaction restriction fragment length polymorphism and analyzed in relation to children with influenza.Results:Compared with the healthy control group[(109.65±4.35) nmol/L], the serum 25-OH-D levels in the study groups were lower[(73.55±2.46)nmol/L in the mild group, (45.59±4.62) nmol/L in the severe group, and (33.65±3.87) nmol/L in the critical group]( P<0.05); Genotypes AA, Aa and allele A of the ApaI locus(51.74%, 22.67%, and 63.08%, respectively)and genotypes FF, Ff and allele F of the FokI locus(41.86%, 34.88%, and 59.30%, respectively)accounted for a significantly higher proportion of cases in the study group than those in healthy control group(11.36%, 14.77%, 18.75%, 10.23%, 13.64%, and 17.05%, respectively)( P<0.05). The proportion of allele A at the ApaI locus and genotypes AA and Aa in severe group(63.70%, 43.84%, and 28.76%) were significantly higher than those in mild group(47.37%, 35.09%, and 24.56%)( P<0.05); The proportion of allele A and genotype AA and at the ApaI locus in critical group(92.86%, 88.10%, and 49.52%) were significantly higher than those in severe group( P<0.05). Serum 25-OH-D<50 nmol/L( OR=5.087, 95% CI 3.114-5.648), ApaI site genotypes AA ( OR=4.011, 95% CI 1.217-18.624)and Aa( OR=3.839, 95% CI 2.483-1.456), FokI site genotypes FF( OR=4.112, 95% CI 3.215-20.775)and Ff( OR=4.591, 95% CI 0.032-10.936)were risk factors for the onset of influenza in children. Conclusion:Serum 25-OH-D deficiency is associated with childhood influenza, and VDR gene genotype AA and Aa of ApaI locus, and FokI site genotype FF, Ff may increase the risk of childhood influenza susceptibility, and allele A of ApaI locus and genotypes AA and Aa are associated with the severity of influenza.

Objective:To explore the expression of signal sequence receptor subunit 1 (SSR1) and its prognostic value in hepatocellular carcinoma.Methods:Search the expression data and relevant clinical data of SSR1 in hepatocellular carcinoma patients from the Cancer Genome Atlas (TCGA) database to June 20, 2021, and download relevant public data. The expression levels of SSR1 in 334 cases of hepatocellular carcinoma with complete information and data were analyzed retrospectively. The expression difference of SSR1 gene between hepatocellular carcinoma and adjacent tissues was analyzed by Wilcoxon signed rank test. Patients with hepatocellular carcinoma were divided into high expression group and low expression group based on the median value of SSR1 expression level (14.660). χ 2 test was conducted to analyze the relationship between SSR1 expression and clinicopathological features. Cox regression and Log-rank survival test were used to analyze the relationship between SSR1 gene expression, clinicopathological features and overall survival rate in patients with hepatocellular carcinoma. Univariate and multivariate Cox regression analysis were used to determine the factors affecting prognosis. Gene set enrichment analysis (GSEA) was used to predict the possible regulatory pathways. Result:Bioinformatics analysis based on TCGA database showed that the expression level of SSR1 in hepatocellular carcinoma (16.320±7.231) was significantly higher than that in normal liver tissue (7.473±1.410). The difference between groups was statistically significant ( t=8.621, P<0.001).The overall survival rate of patients with high SSR1 gene expression group was lower than that of patients with high SSR1 gene expression group (χ 2=10.1, P<0.001). The high expression of SSR1 gene was related to sex (χ 2=4.392, P=0.036), Stage (χ 2=6.264, P=0.012), T stage (χ 2=4.561, P=0.033) and Grade classification (χ 2=14.015, P<0.001). Multivariate Cox regression analysis showed that patients with high expression of SSR1 gene got worse risk of death ( HR=1.030, 95% CI:1.002-1.060, P=0.036), and SSR1 gene expression was an independent predictor of hepatocellular carcinoma. Gene set enrichment analysis showed that the high expression of SSR1 was related to ubiquitination, cell cycle, RNA degradation, mTOR signal pathway, Wnt signal pathway and MAPK signal pathway. Conclusion:SSR1 gene is significantly up-regulated in hepatocellular carcinoma, which is related to gender, Stage, T stage and Grade classification. Ubiquitination, cell cycle, RNA degradation, mTOR signal pathway, Wnt signal pathway and MAPK signal pathway may be the key pathways for SSR1 to promote the occurrence and development of hepatocellular carcinoma.

Objective:To explore the relationship of Vitamin A and Vitamin D with the incidence and severity of hand, foot and mouth disease(HFMD) as well as with the anti-viral immune index interferon-α(INF-α), and to investigate the role of Vitamin A and Vitamin D in HFMD.Methods:A total of 305 children with Coxsackie virus A6(CA6) HFMD admitted at Xi′an Children′s Hospital from January 2017 to December 2018 were enrolled in the study.One hundred healthy children whose gender and age matched with those of children in the case group were selected as the healthy control group.Serum Vitamin A levels were detected by high performance liquid chromatography.Enzyme-linked immunosorbent assay was used to detect the levels of Vitamin D and IFN-α, and the correlation of the levels of Vitamin A and Vitamin D with the severity of HFMD was analyzed.Results:The levels of serum Vitamin A[(0.96±0.39) mg/mol] and Vitamin D [(42.14±15.13) μg/L] in patients with CA6 HFMD were lower than those of the healthy control group[(1.26±0.29) mg/mol, (49.63±8.86) μg/L], and the differences were statistically significant(all P<0.05). Logistic multivariate regression analysis showed that WBC>15×10 9/L, blood sugar>8.3 mmol/L, the deficiency of Vitamin A level and Vitamin D level were all risk factors for severe CA6 HFMD in children( OR=2.303, 4.622, 7.346, 5.211; all P<0.05). According to the receiver operating characteristic curve analysis, the Youden index was the largest at a Vitamin A level of 0.725 mg/mol, and the corresponding sensitivity and specificity were 82.0% and 64.8%, respectively.When Vitamin D level was 32.88 μg/L, the Youden index was the highest, and the sensitivity and specificity were 84.5% and 61.9%, respectively.The serum IFN-α concentration of patients with CA6 HFMD [(84.44±26.28) ng/L] was higher than that of the healthy control group [(36.58±14.39) ng/L], and the difference was statistically significant( P<0.05). In addition, the serum IFN-α concentration in severe HFMD children [(71.48±18.34) ng/L] was significantly lower than that in the common HFMD children [(91.25±27.27) ng/L], and the difference was statistically significant( P<0.05). The results of correlation analysis showed that serum IFN-α concentration is positively correlated with Vitamin A and Vitamin D levels ( r=0.783, 0.239; all P<0.001). Conclusions:The levels of serum Vitamin A and Vitamin D decreased in children with HFMD.WBC>15×10 9/L, blood sugar>8.3 mmol/L, the deficiency of Vitamin A level and Vitamin D level are related to severe HFMD.The se-rum IFN-α concentration is positively correlated with the levels of Vitamin A and Vitamin D. The deficiency of Vitamin A and Vitamin D is one of the early warning factors of severe HFMD.