ObjectiveTo conduct a systematic comparative study on wild and cultivated Codonopsis pilosula（CP） from three aspects， including characters， microscopy， and contents of primary and secondary metabolites. MethodWild and cultivated CP samples were collected， their characters were measured using vernier caliper， tape measure and balance， the paraffin sections were stained with safranin-fixed green dyeing， and their microstructure were observed under the optical microscope. The content of alcohol-soluble extracts in wild and cultivated CP was determined according to the method for determination of extract under CP in the 2020 edition of Chinese Pharmacopoeia， the starch content was determined by anthrone colorimetry， the content of total polysaccharides was determined by kit method， Fiber analyzer was used to determine the content of fiber components， and ultra performance liquid chromatography（UPLC） was used to determine the content of monosaccharides， disaccharides and some secondary metabolites. Multivariate statistical analysis methods such as principal component analysis（PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA） were employed to screen key differential components between wild and cultivated CP on the basis of variable importance in the projection（VIP） value>1 and P<0.05. ResultIn terms of morphological characteristics， the "lion's head-like" shape， longitudinal wrinkles， and circumferential wrinkles below the root cap of wild CP were more pronounced in wild CP compared to the cultivated ones. Regarding transverse sectional features， wild CP had more fissures on the outer side of the cortex and a larger duramen. Under microscopic examination， wild CP had more stone cells， a larger proportion of xylem， and the presence of cork cells arranged in rings in the xylem， while cultivated CP has a larger proportion of phloem， smaller vessel diameters， and a more loosely arranged vascular system. In terms of primary metabolites， the contents of 45% ethanol-soluble extract and total polysaccharides in cultivated CP were significantly higher than those in the wild ones（P<0.05）， the contents of lignin， hemicellulose， cellulose， fructose and glucose in wild CP were significantly higher than those in the cultivated ones（P<0.05）， while sucrose content in the cultivated CP was significantly higher than that in the wild ones（P<0.05）. Concerning secondary metabolites， the contents of tryptophan and tangshenoside Ⅰ in cultivated CP were significantly higher than those in the wild ones（P<0.05）， whereas the contents of lobetyolinin， lobetyol and atractylenolide Ⅲ in wild CP were significantly higher than those in the cultivated ones（P<0.05）. ConclusionThere are significant differences between wild and cultivated CP in terms of morphological characteristics， microscopic features and chemical composition. Glucose， fructose， sucrose， tangshenoside Ⅰ， tryptophan and cellulose components are the key differential components between wild and cultivated CP. Wild CP contains more polyacetylenes and fructose， whereas cultivated CP has higher levels of tangshenoside Ⅰ and sucrose， with noticeably lower cellulose content. These distinctions may be related to their growth conditions， growth years and cultivation techniques. Based on the results of this study， it is recommended to increase polyacetylenes and the content ratio of fructose to sucrose as an indicators to characterize different production methods of CP， in order to guide the high-quality production of CP.

Objective The objective of this study was to investigate the effect of acidic conditions on epithelial mesenchymal transition(EMT) of HepG2 cells.Methods HepG2 cells were cultured under acidic and alkaline conditions to observe the difference of cell morphology.Wound healing assays were performed to detect the migration ability of the two groups.Matrigel invasion assays were used to detect the invasive ability of the cells.The expression of EMT-related genes at mRNA level was detected by RT-PCR.The expression level of EMT-related gene protein was detected by Western blot.Results The morphology of HepG2 cells was changed from epithelium to interstitial type under acid conditions.The migration ability of HepG2 cells under acidic condition was stronger than that of alkaline conditions.The number of HepG2 cells crossing Matrigel was higher than that of alkaline condition.The mRNA expression of Vimentin,Slug,Snail and Zeb1 related to EMT and the protein expression of Vimentin and MMP9 related to EMT in HepG2 cells were significantly higher than those under alkaline condition.Conclusion The acid conditions can induce the occurrence of EMT in HepG2 cells.

Objective To investigate the clinical manifestations,pathological features,treatment and prognosis of nongesta-tional choriocarcinomas.Methods The clinicopathological data in 4 cases of nongestational choriocarcinoma were retrospectively analyzed and the relevant literatures were reviewed.Results All 4 patients had no confirmed diagnosis at first visit to hospital.All patients received the operative therapy and postoperative bleomycin,etoposide and cis-platinum (BEP)regimen chemotherapy.The followed up lasted for 3-32 months.One case died at postoperative 26 months due to pulmonary metastasis;one case was loss of follow up;two cases had relatively favorable condition.Conclusion Nongestational choriocarcinoma is a rare and highly invasive trophoblastic neoplasm,and is prone for early distant metastasis.The treatment is recommended to adopt operation combined with postoperative combined chemotherapy of multiple drugs.

OBJECTIVETo investigate the association of SNP of CD40 gene and its serum levels with ischemic stroke (IS).

METHODSA total of 202 IS patients from a hospital of Baise city were enrolled in case group from May 2013 to November 2014. At the same time, 109 healthy people who had physical check-ups in the outpatient department at the same hospital were enrolled in the control group. All participants were from Guangxi Zhuang Autonomous Region and unrelated to each other. 3 ml venous blood were collected on the premise of informed consent. The single nucleotide polymorphisms of CD40 gene rs1883832 C/T, rs13040307 C/T, rs752118 C/T and rs3765459 G/A were analyzed using a Snapshot SNP genotyping assays, and the serum levels of CD40 were tested by ELISA. t-test was used to compare the serum levels of CD40 between the case and control group, and the genotypes at different locuses in case group; χ(2) test was used to compare the distribution differences of the CD40 gene locuses in different genotypes and allele between the case group and the control group; alleles was established as independent variables, the occurrence of the IS as dependent variable, and expressed relative risk with OR (95%CI) value.

RESULTSIn the case group, the frequency of CC, CT and TT genotypes in CD40 gene rs1883832 C/T were 21.78% (44/202), 49.51% (100/202) and 28.71% (58/202), respectively, and 33.17% (66/199), 48.74% (97/199), 18.09% (36/199) in the control group, respectively, the differences between the two groups was significant (χ(2)=9.57, P=0.008). The CD40 serum levels were (62.7 ± 24.5) pg/ml in the case group, which was higher than that in the control group (45.3 ± 17.2) pg/ml (t=8.97, P<0.001). The serum levels of TT and CT genotypes in CD40 gene were (65.9 ± 26.3) and (64.3 ± 25.9) pg/ml, respectively, and the differences were significant when comparing with CC genotype (t equaled 5.34 and 5.03, respectively, P<0.001). The risk of developing IS was 1.56 times higher in 1883832 T allele carriers than that in rs1883832 C allele carriers (OR=1.56, 95% CI: 1.18-2.06); Combined genotype analysis displayed that CD40 gene rs1883832 C/T, rs13040307 C/T, rs752118 C/T and rs3765459 G/A polymorphisms showed strong linkage disequilibrium, the case group TCCA haplotype was tested to be associated with a significantly increased risk of IS as compared with that in the control group(OR=2.49; 95%CI: 1.13-5.48).

CONCLUSIONCD40 gene rs1883832 C/T polymorphism and its TCCA haplotype were possibly associated with ischemic stroke, and the susceptibility gene for ischemic stroke may be rs1883832 T allele.

Alleles ; CD40 Antigens ; blood ; genetics ; Case-Control Studies ; Cell Differentiation ; China ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Haplotypes ; Humans ; Polymorphism, Single Nucleotide ; Stroke ; blood ; genetics

Objective To explore the association of CD40 gene single nucleotide polymorphisms (SNPs) and haplotypes with the susceptibility to systemic lupus erythematosus (SLE),as well as the association of serum levels and genotypes of CD40 with the occurrence of SLE.Methods A multiplex PCR single-base extension assay (PCR-SBE) and DNA sequencing were performed to analyze 4 SNPs of the CD40 gene,including rs1883832 C/T,rs13040307 C/T,rs752118 C/T and rs3765459 G/A,in 205 patients with SLE (SLE group) and 220 healthy human controls (control group).Enzyme-linked immunosorbent assay (ELISA) was conducted to measure serum levels of CD40 in these subjects.Results Compared with the control group,the SLE group showed significantly increased serum levels of CD40 (P ＜ 0.05).There were significant differences in genotype and allele frequencies of the SNP rs1883832 C/T in the CD40 gene between the SLE group and control group (all P＜ 0.01).Relative risk analysis showed that the risk of developing SLE in rs1883832 T allele carriers was 1.517 times that in rs1883832 C allele carriers (OR =1.517,95％ CI:1.157-1.990,P=0.003).Moreover,serum levels of CD40 were significantly higher in rs1883832 T allele carriers than in rs1883832 C allele carriers (P ＜ 0.01).The risk of developing SLE was significantly increased in TCCA haplotype carriers compared with the healthy controls (OR =2.322,95％ CI:1.181-4.564,P=0.012).Conclusion The CD40 gene rs1883832 C/T polymorphism and its TCCA haplotype were both associated with the occurrence of SLE,and the rs1883832 T allele may be a gene predisposing to SLE.

Objective to provide the evidence for inducing the SAP model in rats with proper concentration of sodium taurocholate.Methods 60 SD rats were divided into sham operated group, group of 1.5% in concentration, group of 3.5% in concentration and group of 5% in concentration randomly, while the SAP model was induced by the sodium taurocholate concentration of 1.5%,3.5% and 5% with the method of retrograde injection into the biliopancreatic duct.To calculate the mortality of different groups, measure the serum amylase, tumor necrosis factor -α(TNF -α) and interleukin -6 (IL -6),and to observe the pancreatic pathological scores of HE staining in rats.Results The mortality in group of 5% in concentration has a significant ascending compared with group of 1.5% in concentration, while the serum amylase, tumor necrosis factor -α(TNF -α) , interleukin -6( IL -6), pathological score of hemorrhage and acinar necrosis in group of 5% in concentration have a significant ascending compared with group of 1.5% in concentration and group of 3.5% in concentration.Conclusions A better SAP model may be induced by sodium taurocholate with the concentration of 5% by the method of retrograde injection into the biliopancreatic duct, which may accord with the physiological and pathological manifestation of SAP.

Objective To develop an immunomagnetic bead ( IMB) assay for quantitative detection enolase ( Eno) of Candi-da albicans, and to improve the diagonosis of invasive candidiasis . Methods The immunomagnetic bead was prepared by conjuga-ting with Anti Eno of Candida albicans monoclonal antibody .HRP-conjugated goat polyclonal antibody against Candida albicans Eno was employed as detecting antibody .The performance parameters of the IMB assay including precision , specificity, linear range and limit of detection were verified by using recombinant Candida albicans Eno.Then the developed assay was applied to determine Eno levels in supernatant of pathogenic fungi cultures . Results The intra and inter-coefficient of variation was 4.54%, 5.87% and 5.26%, 8.82%at the concentration of 25 ng/mL and 5 ng/mL, respectively.The limit of detection was 0.5 ng/mL.The linear range was(0.5-50) ng/mL.The level of Eno in Candida albicans culture after incubated in 37℃for 24 h was 3.89 ng/mL and gradually in-creased to 37.89 ng/mL at 120 h.There was a positive correlation between the level of Eno and growth hyphae of Candiad albicans. There was weak cross reaction with Candida parapsilosis and no cross reaction with Candida tropicalis, Candida guilliermondii, Candida glabrata, Cryptococcus neoformans and Saccharomyces cerevisiae. Conclusion An IMB assay for quantitative detection Eno of Candida albicans was developed , which was more sensitive , rapid and reliable than previous qualitative ELISA .The IMB assay has the potential to be applied to the research in invasive candidasis .

Objective To study the effect of Mycoplasma pneumoniae infection on the coagulation system in children and to clarify its mechanism in order to guide the therapeutic strategy for Mycoplasma pneumoniae infection in clinical practice.Methods A prospective and case control study was carried out in 36 children with Mycoplasma pneumoniae infection in Weifang People ' s Hospital from January through December in 2011.The 36 patients were divided into two groups according to the MP-IgM plasma titers,namely low-titer group with MP-IgM 1 ∶ 80-1 ∶ 160 (n =17) and high-titer group with MP-IgM 1 ∶ 320-1 ∶1280 (n =19).The subjects of control group (n =20) were recruited from healthy children as they took routine physical examination during the same period.Six indexes related to the coagulation function as well as D-dimer in plasma were determined.All data were analyzed by using SPSS 15.0 software.Results The levels of fibrinogen (FIB) and D-dimer in the low-titer group and high-titer group were significantly higher than those in the control group (P ＜ 0.05).The prothrombin (PT) and activated partial thromboplastin time (APTT) in the high-titer group were significantly shorter than those in the control group respectively (P ＜ 0.05).Conclusions Mycoplasma pneumoniae infection in children gives rise to the potential for activating the extrinsic and intrinsic coagulation system,promoting thrombosis and in turn inducing ischemic stroke in serious cases.

Objective To study the frequencies of allele and genotype distribution of alpha-B-crystallin (CRYAB ) gene rs3212227 and rs6894567 single nucleotide polymorphism (SNP) in Chinese guangxi populations ,and to Compare the distribution differences among different ethnic .Methods The CRYAB gene rs3212227 and rs6894567 polymorphisms were detected by the pol-ymerase chain reaction-single base extension (PCR-SBE) technique and DNA sequencing methods in 199 Chinese guangxi popula-tions ,frequencies of allele and genotype of CRYAB gene SNP loci ,rs3212227、rs6894567 were analyzed in guangxi populations com-pared with other the four populations (HapMap-CEU ,HapMap-YRI ,HapMap-JPT and HapMap-HCB) from Human Genome Pro-ject group (Hapmap) data .Results There were CRYAB gene polymorphisms in Guangxi populations .The frequencies of allele and genotype distribution of CRYAB gene rs3212227、rs6894567 polymorphisms had significant difference compared with HapMap-CEU and HapMap-YRI populations (P<0 .05) ,and had no significant difference compared with HapMap-JPT and HapMap-HCB (P>0 .05) .Conclusion The frequencies of allele and genotype distribution of CRYAB gene rs3212227、rs6894567 polymorphisms are significantly difference compared with others ethnic populations ,and this variation might account for a variety of clinical mani-festation and morbidity of of some CRYAB related diseases .

Objective To detect quantitatively AKAP12 methylation and evaluate its clinical significance in peripheral blood in colorectal cancer. Methods MS-HRM technology was used to detect quantitatively AKAP12 methylation in peripheral blood from 80 colorectal cancer patients and 20 healthy volunteers. They also validated the reproducibility and compared with MSP. Results Thirty-eight of the 80 colorectal cancer samples (47. 5% ) were found to be methylated at the AKAP12 promoter region by MS-HRM (the methylation levels of 24 cancer samples ranged between 1 % and 20% , the methylation levels of 12 cancer samples ranged between 20% and 60% , the methylation levels of 2 cancer samples ranged between 60% and 100% ). The methylation levels of 2 health samples were less than 10% . They also compared the results generated by MS-HRM with a traditional MSP assay. The AKAP12 MS-HRM assay was able to reproducibly detect 1% AKAP12 methylated DNA, whereas the MSP method was unable to detect less than 10% methylation. No significant correlation was observed between the AKAP12 methylation levels and patients' age and gender. However, AKAP12 methylation was significantly higher in DNA from colorectal cancer patients with high Dukes stage and differentiation (x2 =5. 93 or 8. 41, P = 0.01). Conclusions The authors demonstrate here for the first time, the utility of quantitative AKAP12 MS-HRM analysis of promoter methylation in peripheral blood samples. AKAP12 MS-HRM quantitative methods have many promising applications in the detection of colorectal cancer.