Objective To assess the application of ultrasonography in prenatal diagnosis of fetal choledochal cysts.Methods The clinical data of 25 fetuses,who were diagnosed as fetal choledocho cysts using ultrasonography and followed up,were retrospectively analyzed.Results Among 25 cases,24 cases were confirmed as congenital choledochal cysts after normal delivery or abortion,and 1 case of liver cyst was misdiagnosed.The 24 cases' prenatal ultrasound showed liver cystic space occupying wihich was connected to intrahepatic bile duct or gallbladder.The cysts wall was thin,and the colour Doppler flow imaging CDFI showed that blood vessels circled around the cyst.In 24 fetuses with prenatal choledochal cysts,21 were fullterm birth,3 were aborted including 1 case with other congenital abnormalities.The size of choledochal cysts was not changed in 5 cases,and was enlarged with the gestational ages in remaining 16 cases fetuses.The choledochal cysts did not affect fetal growth and development.The newborn infants with congenital choledochal cysts were treated surgically within 8 months after birth and all recovered well.Conclusion Fetal choledochal cysts have typical sonographic manifestations.Ultrasonography can be used for prenatal diagnosis of congenital choledochal cysts,which can be successfully treated by surgery early after birth.

Objective To study the effect of thyroid-stimulating hormone (TSH) on human vascular endothelial cells and smooth muscle cells and to explore the roles of TSH in the development of atherosclerosis.Methods Human vascular endothelial cells and smooth muscle cells were cultured in vitro.MTT method was used to assay the effect of TSH on cell viability.Real-time PCR was used to determine the levels of endothelial nitric oxide synthase (eNOS),prostacyclin(PGI2),endothelin-1 (ET-1),plasminagen activator inhibitor(PAI-1) (mRNA) in endothelial cells and the phenotype transition of smooth muscle cells.The effect of TSH to the cycle of smooth muscle cells was detected by using flow cytometry.Immunocytochemistry and Western blot were used to investigate the expression of the cell cyclin A,D1,and the expression of endothelial cell associated factors eNOS and ET-1.Results Compared with the control group,eNOS and PGI2 mRNA levels decreased while ET-1 and PAI-1 mRNA levels increased when different concentrations of TSH were applied to endothelial cells(P＜0.05).The level of eNOS protein was decreased gradually while the level of ET-1 protein was gradually increased(P＜0.05).Different concentrations of TSH applied to smooth muscle cells could promote the transition of cell cycle phase G2 to phase M and increase the expression of cell cyclin A and D1.Conclusion TSH may damage the function of vascular endothelial cells and promote the proliferation of smooth muscle cells.

Objective To investigate the effect of calcipotriol on melanin synthesis by human melanocytes and its possible action mechanism.Methods Primary melanocytes were cultured with various concentrations(10~(-5),10~(-6),10~(-7),10~(-8),10~(-9),10~(-10) mol/L)of calcipotriol for 24 or 48 hours.Subsequently,MTT assay,NaoH assay.Dopa-oxidase assay,Western blot and semiquantitative RT-PCR were used to measure the cell proliferation of,melanin synthesis by.tyrosinase activity,protein and mRNA expression levels in the melanocytes.respectively.Those untreated melanocytes served as the control.Results The calcipotriol between 10~(-9) and 10~(-5) mol/L had no significant effect on the proliferation of cultured melanocytes(P＞0.05).while that of 10~(-9) and 10~(-8) mol/L increased tyrosinase activity by 137%and 123%,and enhanced melanin synthesis by 40.63%and 18.75%,respectively,compamd with untreated melanocytes(both P＜0.05).Moreover,the tyrosinase protein level increased by 270.4%(P＜0.05)in melanocytes treated with calcipotriol at 10~(-9) mol/L for 24 hours.The strongest tyrosinase activity and melanin synthesis was observed in melanocytes treated with calcipotriol of 10~(-9) moI/L.Conclusions The proliferation of melanocytes is unaffected by calcipotriol at 10~(-9) to 10~(-5) mol/L,but it can elevate the expression of tyrosinase protein,enhance tyrosinase activity,and promote melanin synthesis in melanocytes.

Objective To test the technical param eters of GlobalFiler?PC R A m plification K it for its ap-plication to forensic application value and to investigate the genetic polym orphism s. Methods The valida-tion w as conducted in sensitivity, m ixed sam ples, species specificity, adaptability, survivability, consistency, peak height balance and stability. The am plification and detection of the genom ic D N A from 373 unre-lated individuals from B eijing H an nationality w ere extracted by autom ation w orkstation. Results Global-Filer?PC R A m plification K it w as adaptive to som e m ixed, degraded and inhibited sam ples. The pow er of sensitivity and adaptability and peak height balance show ed w ell. The distributions of genotype fre-quencies for 21 STR loci in the population w ere all in accordance w ith H ardy-W einberg equilibrium (P>0.05). The PIC value of the 21 STR loci w as am ong 0.536 to 0.940; the H value w as am ong 0.558 to 0.933; the D P value w as am ong 0.783 to 0.992; the PE value w as am ong 0.243 to 0.874. Conclusion GlobalFiler?PC R A m plification K it is suitable for crim inal cases and D N A database in forensic practice. A nd 21 STR loci in B eijing H an nationality have high polym orphism , w hich have ap-plication value in forensic practice and population genetics.

This study was aimed to establish a method for the rapid content determination of Ixerin Z and 11,13α-dihydroixerin Z in Ku-Die-Zi (KDZ) injection by UPLC-ESI-MS/MS. The separation was performed on a Waters ACQUITY BEH C18 column (2.1 mm × 50 mm, 1.7μm) by using a gradient elution with the mobile phase of acetonitrile-water at the flow rate of 0.4 mL·min-1. The column temperature was set at 40℃. Multi-reaction moni-toring (MRM) scanning was employed for quantification in ESI negative mode. The results showed that two sesquit-erpene lactones in KDZ injection were totally separated within 2 min. The linear range of Ixerin Z was 5.70-182.50 ng·mL-1, and the linear range of 11,13α-dihydroixerin Z was 4.60-131.25 ng·mL-1. The correlation coefficient r was more than 0.999 0. The recovery rates (n = 6) were 98.18% and 97.52%, with RSDs < 1.5%. The established method was successfully applied for simultaneous content determination of Ixerin Z and 11,13α-dihydroixerin Z in 6 batches of KDZ injection from 2 factories, which had some variations on the content determination results. It was concluded that the method was rapid, accurate and sensitive, which can be used for the content determination of two sesquiterpene lactones in KDZ injection.

OBJECTlVE To study the anti-HBV activity of prepared recombinant human serum aIbu-min-interferon α-2b fusion protein(HSA-IFNα-2b) in vitro. METHODS HepG2 ceIIs were infected with recombinant adenovirus with green fIuorescence protein and 1.6-foId HBV DNA(AdGFP-HBV). The ex-pression of HBV antigens,HBsAg and HBeAg in cuIture medium was detected by ELISA assay. The tox-icity of HSA-IFNα-2b on HepG2 ceIIs was evaIuated by mTT assay.The reIative expression of HBV RNA in ceIIs and the absoIute quantity of HBV DNA in cuIture supernatant were determined by quantitative PCR assay. The activity of HBV enhancer Ⅰ was detected by DuaI-Reporter gene assay. RESULTS HBV couId repIicate and express in HepG2 ceIIs after infection with AdGFP-HBV. The expression of HBsAg and HBeAg in cuIture serum of HepG2 ceIIs infected with AdGFP-HBV decreased by 51.32%(P﹤0.01)and 50.26%(P﹤0.01),respectiveIy,when HSA-IFNα-2b 500 kU·L-1 was added. The same concentration of HSA-IFNα-2b didn't inhibit the proIiferation of HepG2 ceIIs,but inhibited HBsAg in a concentration-dependent manner. The regression formuIa between HBsAg inhibitory rate(Y)and con-centration of HSA-IFNα-2b(X)was Y=21.11 IgX+11.91(r 2 = 0.954),IC50 = 63.76 kU·L-1 . HBV RNA in ceIIs and HBV DNA in the cuIture serum decreased by 52.83%(P﹤0.01)and 53.07%(P﹤0.01), respectiveIy,when HSA-IFNα-2b 500 kU·L-1 was added. The activity of enhancer Ⅰ decreased by 40.04%(P﹤0.01)when HSA-IFNα-2b 500 kU·L-1 was added. CONCLUSlON The ceII modeI of HBV repIication for evaIuating anti-HBV agents is successfuIIy estabIished. HSA-IFNα-2b exhibits noticeabIe anti-HBV effect invitro.

Mucin-type O-glycosylation is one of the most common post-translational modifications in proteins,capable of altering protein conformation and biological functions.It plays a crucial role in biological processes such as cell signaling,cell adhesion,and immune responses.Polypeptide N-acetylgalactosaminyltransferase 3(GALNT3),as the initiating enzyme of mucin-type O-glycosylation,is of paramount importance in maintaining the homeostasis of human cells and tissues.Dysfunction of GALNT3 has been found to play a role in various diseases,such as calcium-phosphorus metabolism disorders and atherosclerosis.Additionally,GALNT3 is abnormally expressed in several types of tumors,including colorectal cancer,lung cancer,and ovarian cancer.Its expression is associated with the clinical pathological features of patients and poor prognosis,making it a potential biomarker for early tumor diagnosis and prognosis evaluation.Further research shows that GALNT3 can both regulate glycosylation levels to reduce adhesion between tumor cells and activate multiple metabolism-related pathways,promoting tumor cell invasion and metastasis.This review summarizes the role of GALNT3 in the development of malignant tumors and discusses the prospects and challenges of developing anti-tumor drugs targeting GALNT3.

Objective To study the plasma D-dimer (DD) level changes in the patients with rheumatoid arthritis (RA), and to analyze its correlation with various clinical indicators. Methods Selected a total of 138 cases of RA patients and 50 cases of normal person as matched group the plasma D-dimer level and the correlation analysis with age, rheuma-toid factor (RF), erythrocyte sedimentation rate (ESR), visual analogue scale (VAS score)and platelet were analyzed. Results The level of D-dimer level in RA patients was significantly higher than that of healthy controls, and the dif-ference was statistically significant (P<0.01);The D-dimer level of high disease activity group and low disease activity group with RA patients were significantly higher than the remission groups of RA patients (P all <0.01); The D-dimer level in high disease activity group of RA patients was significantly higher than the low disease activity group (P<0.01); There were positive correlations between the D-dimer level and the age, ESR, VAS score, RF, platelet in all RA patients (P all <0.01). Conclusion There is high coagulation state in RA patients, so the D-dimer can be used as a clinical nonspecific inflammatory reaction index in patients with RA, and can direct the clinical treatment.

Objective To explore a method for assessing the irregular small opacities profusion associated with occupational pneumoconiosis in chest computed tomography (CT). Methods A total of 20 occupational pneumoconiosis patients whose primary manifestation was irregular small opacities on chest digital radiography (DR) were collected as the research subjects using a retrospective study method. Comparative analysis was performed between chest DR and five mm coronal multi-planar reconstruction (MPR) of chest CT images to identify the causes of irregular small opacities. An evaluation method for the profusion of associated images of irregular small opacities in chest CT was established using technique for order preference by similarity to ideal solution-analytic hierarchy process (TOPSIS-AHP), and the results were compared against GBZ 70-2015 Diagnosis of Occupational Pneumoconiosis. Results The abnormal image distribution on the five mm coronal chest CT MPR images of the 20 patients was as follows: three cases of high-density small circular opacities, seven cases of low-density circular small opacities, six cases of diffuse low-density ground-glass opacities (GGO), four cases of reticular opacities, three cases of plate-like GGO, three cases of honeycomb opacities, and four cases of increasing lung texture. The CT values of abnormal images, from high to low were: honeycomb opacities > plate-like GGO > low-density circular small opacities > diffuse low-density GGO (all P<0.05). The consistency test results indicated that the evaluation method for the profusion of associated images of irregular small opacities in chest CT showed high level of agreement with the profusion determination criteria outlined in GBZ 70-2015 Diagnosis of Occupational Pneumoconiosis (Kappa=0.78). Conclusion Irregular small opacities observed on chest DR are formed by the superposition of multiple images of abnormal pulmonary fibrosis in patients with occupational pneumoconiosis. TOPSIS-AHP can be used to establish an evaluation method of the profusion of associated image of irregular small opacity in chest CT.

OBJECTIVETo construct a dual-reporter gene system that will be applicable for use as a tool to screen and evaluate therapeutic drug compounds that inhibit transcription of the gene encoding collagen I, chain at1 (COL1A1).

METHODSThe full-length eDNA of transforming growth factor beta1 (TGFbeta1) was cloned by RT-PCR and inserted into two vectors, pcDNA3.1 and pJW4303, for construction of two eukaryotic expression vectors, pcDNA3.1-TGFbeta1 and pJW4303-TGFbeta1.Next, the promoter region of COL1A1, cloned by PCR using human genome DNA as template, was inserted into the vector pGL4.29 to construct the reporter gene vector, pGL4.29-COL1A1 promoter.All three recombinant vectors were verified by restriction enzyme digestion and DNA sequencing.Either the pcDNA3.1-TGFbeta1 or pJW4303-TGFbeta1 vector along with the pGL4.29-COL1A1 promoter vector or a pRL-null, control reporter, vector were co-transfected into the LX-2 human hepatic stellate cells to establish the transcription-activated dualreporter gene system.This system was used as a cell model for screening anti-liver fibrosis compounds that inhibit the transcription of COL1A1.Dexamethasone, a model drug that is known to inhibit the expression of COL1A1, was used as a control to validate the dual-reporter gene system.

RESULTSThe two TGFbeta1-expressing vectors and the reporter gene vector containing the promoter region of COL1A1 were successfully constructed.The results of a dual-reporter gene assay showed that TGFbeta1 co-expression increased the activity of the COL1A1 promoter by above 200-fold (t =21.78, P =0.0001), whereas in the absence of TGF31 co-expression the activity was below 2-fold (t =3.396, P =0.0274).The transcriptionactivated dual-reporter gene system was successfully established.The model drug, dexamethasone, effectively inhibited the activity of the COL 1A1 promoter in dose-dependent manner; the activity decreased 29.6% with 10 mumol/L dexamethasone (t =4.140, P =0.0144) and 53.9% with 100 mumol/L (t =6.193, P =0.0035).

CONCLUSIONThe dual-luciferase reporter system of TGFbeta1 and COL1A1 co-expression developed here can be used as a cell model to screen and evaluate anti-liver fibrosis compounds that inhibit activity of the COL1A1.

Base Sequence ; Collagen Type I ; genetics ; Drug Evaluation, Preclinical ; Genes, Reporter ; Genetic Vectors ; Humans ; Liver Cirrhosis ; drug therapy ; Luciferases ; Promoter Regions, Genetic ; Transcriptional Activation ; drug effects ; Transfection ; Transforming Growth Factor beta1