Fufang Ganning(FGN)is an oral liquid,consisting of 8 kinds of chinese herbs including Radix Astragali,Radix Salviae Multiorrhizae,Radix Curcumae,Rhizoma Polygonati,etc.It was found to be effective in protecting against experimental acute hepatic damage caused by CC14,TAA and D-Glan in mice,FGN could reduce ALT and retention of BSP,and histopathological chan- ges revealed that the hepatic injury induced by CC14 in animals received FGN was much less severe than those of the control ones.It could also lower ALT and serum bilirubin in mouse and rat models with hepatic injury caused by ?- isothioeyanate.On the other hand,in the animal models with cirrhosis induced by CC14,FGN could not only cut down ALT but also lessen the degeneration and necrosis of hepatic cells and cirrhotic changes.Meanwhile,FGN could significantly increase the clearance index of carbon particles(phagocytic func- tion)and serum hemolysin in normal mice.It is concluded that FGN has the effects of protecting liver from damage and enhancing the immune function.

Objective To describe and analyze the clinical characters of patients with FRG from 7 Chinese families.Then analyze and identify their mutations in SGLT2 gene,and explore the association of genotype and phenotype.Methods Quantitative test for 24-hour urine glucose and other laboratory tests were carried out among 7 probands (14 patients in all) and their family members from 7 pedigrees (totaling 23 subjects).All coding regions,including intron-exon boundaries,were analyzed using PCR followed by direct sequence analysis.Results Five novel mutations in SLC5A2 gene were identified in this investigation,including four missense mutations (A Serine to Glycine at position 335 (c.1003A＞G,p.S335G),a Glutamine to Arginine at position 448 (c.1343A ＞ G,p.Q448R),an alanine to proline at position 474 (p.A474P,c.1420G ＞ C) and a glycine to aspartic acid at position 580 (c.1739G ＞ A,p.G580D) and a deletion in intron 7 (c.886(-10_-31)del).By the minigene studies using the pSPL3 plasmids,we confirmed the deletion c.886(-10_-31)del as a splicing mutation.In this study,the mutation c.886(-10_-31)del accounted for about 43％ of the total alleles (12/28).These patients with compound heterozygous or homozygous mutations manifested middle degree or severe glycosuria (Quantitative test for 24-hour urine glucose:10.56-50.68 g/1.73 m2),however those with heterozygous variants presented with mild to moderate glycosuria (Quantitative test for 24-hour urine glucose ≤ 2.45 g/1.73 m2).This fits co-dominant inheritance pattern.Conclusions Five novel mutations which may be related to FRG are found in this study,and c.886(-10-31) del may be a high frequency mutation in Chinese patients.

BACKGROUND:Human umbilical cord mesenchymal stem cels (hUC-MSCs) can obviously relieve liver cirrhosis, and thereby repair liver injury. However, the molecular mechanism of hUC-MSCs therapy for liver cirrhosis is limited at present, and especialy the non-coding RNA regulation of hepatic gene changes has not been detailed. OBJECTIVE:To investigate the changes of microRNA after hUC-MSCs therapy in rats with liver cirrhosis. METHODS:Liver cirrhosis models were established in rats using carbon tetrachloride subcutaneous injection plus oral administration of alcohol. At 8 weeks after modeling, hUC-MSCs were injectedvia the tail vein once a week for 4 consecutive weeks. At 1 week after the last injection, rat liver tissues were colected for paraffin embedding. Liver RNA was extracted for gene chip analysis. Blood samples were colected and analyzed using an automatic biochemical analyzer to detect the changes of liver function. RESULTS AND CONCLUSION:Alanine aminotransferase, aspartate aminotransferase and gamma-glutamyl transpeptidase were improved significantly after hUC-MSCs therapy. Fat lesions and necrosis of hepatocytes were significantly reduced. MicroRNA expression microarray hybridization analysis and PCR results showed that rno-miR-369-5p, rno-miR-3584-5p and rno-miR-153* were down-regulated during modeling and increased after hUC-MSCs therapy. And rno-miR-93, rno-miR-199a-3p, rno-miR-195, rno-let-7a and rno-miR-19a were firstly up-regulated in the process of modeling and then down-regulated obviously after hUC-MSCs therapy. These results suggest that hUC-MSCs may reverse liver cirrhosis and liver cel damage through up-regulation of rno-miR-369-5p, rno-miR-3584-5p and rno-miR-153*, and down-regulation of rno-miR-93, rno-miR-199a-3p, rno-miR-195, rno-let-7a and rno-miR-19a.

Lidamycin (LDM) is a potent antitumor antibiotic. Previous studies have shown that LDM could inhibit proliferation and migration in endothelial cells. In the present report, the effect of LDM on angiogenesis of zebrafish embryo was studied. The results showed that treatment of zebrafish embryos with LDM resulted in significant inhibition of angiogenesis. Morphological observation, quantitative endogenous alkaline phosphatase (EAP) assay, alkaline phosphatase staining, and transgenic zebrafish assay were performed to evaluate vascular development defects in zebrafish. The results indicated that after the zebrafish embryos were exposed to LDM, angiogenesis defects of zebrafish embryos were observed, including pericardial edema, reduced numbers of circulating red blood cells, suppression of zebrafish vessel growth, and absences of SIV (subintestinal vein). The expression of VEGF was detected by RT-PCR assay, quantitative reverse transcriptase real-time PCR (qRT-PCR) assay and Western blotting analysis. The results revealed that LDM could inhibit the expression of VEGF protein, while the expression of mRNA was not significantly affected. The study suggests that LDM could inhibit the zebrafish embryo angiogenesis by down-regulation ofVEGF expression.

Objective To screen the 5 EV71 vaccine candidates which were isolated from MRC-5 cells to find one as the vaccine virus. Methods The ICR mother mouse were immunized by intraperitoneal injection with the 5 vaccine candidates which were made from monoclonal EV71 virus. Two weeks after booster immunization, the animals were allowed to mate, another booster was given after 2 weeks, and then attracted the milk mouse within 24 h with different types of virus by cranial cavity injection. The survival condition were recorded everyday, and the antibody titre(IgG) were detected by ELISA, the virus titre of intestine were detected by nest-PCR, and neutralizing antibodies were determined using a microassay with MRC-5 cells, and then the data were analyzed by SPSS16.0. Results The antibody titre of 5 virus immunized ICR mouse were improved with the increase in the immune times, and they got difference in neutralization capacity, the survival rate after fatal attract and the virus titre of the intestine. Conclusion It proved that the five vaccine candidates were different at the molecular level, cellular level and individual level. 123 strain was the best one in immunogenicity and immunoprotective property, which agreed with the vaccine requirement.

AIM:To construct AFT024-SCF cell line and HPC-Lhx2 cell line for confirming the biological function of AFT024-SCF.METHODS:The HPC-Lhx2 cell line, AFT024-SCF cell line and AFT024-GFP cell line were constructed by retro-viral infection.The expression of stem cell factor(SCF) in AFT024-SCF cells was detected by real-time PCR and Western blot.SCF in the supernatant of AFT024-SCF was detected with ELISA.The supernatant of AFT024-SCF and AFT024-GFP were collected and then diluted (1:10) with basic IMDM medium.So we made 4 culture medium:AFT024-SCF medium was used for experiment group, AFT024-GFP medium was used for endogenous negative control, IM-DM basic medium was used for exogenous negative control, and IMDM basic medium with SCF was used for positive con-trol.SCF-dependent HPC-Lhx2 cell line was cultured in these 4 different medium for 72 h.According to MTT method and colony forming experiment, the biological function of AFT024-SCF was confirmed by the proliferation ability of SCF-depend-ent HPC-Lhx2 cell line.RESULTS:SCF was highly expressed in AFT024-SCF cells.After cultured for 72 h, neither IM-DM basic medium nor GFP-AFT024 medium support HPC-Lhx2 cell line proliferation.However, AFT024-SCF medium supported HPC-Lhx2 cell line expansion as well as the positive control medium.CONCLUSION:AFT024-SCF cells ex-press SCF successfully and recombinant SCF can be replaced by the supernatant of AFT024-SCF culture medium for expan-ding HPC-Lhx2 cell line in vitro.

Objective To evaluate the value of neck blood vessel colored doppler ultrasound (NBVCDU) and magnetic resonance angiography (MRA) to extracranial carotid artery stenosis in patients with transient ischemic attack (TIA).Methods After implementing NBVCDU and MRA examinations at the same time,45 TIA patients with at least one examination showing arteriostenosis in extracranial section were chosen to carry out cerebral digital subtraction angiography( DSA ),then the stenosis rate was calculated by American symptomatic carotid endarterectomy trial (NASCET) method.Results Regarding DSA as the gold standard,for 45 TIA patients that having 180 arteriostenosis in extracranial section, sensitivity,specificity,accuracy of NBVCDU examination was 93.51% ,95.15% ,94.44%, Kappa = 0.735; sensitivity,specificity,accuracy of MRA was 92.21% ,94.17% ,93.33% , Kappa =0.681; sensitivity,specificity,accuracy of NBVCDU combined with MPA was 97.40% ,99.03% ,98.33%, Kappa = 0.872.Conclusions The sensitivity and accuracy of arteriostenosis in extracranial section by NBVCDU examination is higher than that by MRA, and it is suitable in the crowd primary examination.NBVCDU combined with MRA has shown good consistence with DSA for diagnosing arteriostenosis in extracranial section,but can't replace DSA comlpetely.

OBJECTIVETo study the chemical constituents from leaves of Uraria lacei.

METHODChemical constituents were isolated by silica gel column and Sephadex LH-20, and identified by physiochemical and spectral analyses and by comparison with the standard compounds.

RESULTEleven compounds were isolated and identified as naringenin-7-0-beta-D-glucopyranside (1), (2S)-5, 7-dimethoxy-4'-hydroxyflavan (2), dalbergioidin (3), 5, 7-dihydroxy-2'-methoxy-3', 4'-methylenedioxyisoflavanone (4), apigenin (5), 5, 7-dihydroxy-2', 4'-dimethoxyisoflavanone (6), 5, 7, 2', 4'-tetrahydroxyisoflavone (7), emodin (8), saliylic acid (9), daucosterol (10), and tetracosane (11).

CONCLUSIONAll compounds were isolated from this plant for the first time.

Objective To analyze the clinical features,diagnosis and treatment of testicular torsion.Methods The clinical data of 95 patients with testicular torsion admitted from March 2001 to April 2017 were analyzed retrospectively.Results The age of patients ranged from 13 to 42 years (mean 19.6 years).Among 95 patients,91 cases had essential clinical symptoms of scrotal pain;1 case reported the right testicular enlargement without pain,and 3 cases presented with left inguinal mass.Color Doppler flow imaging (CDFI) was performed in 91 patients,reduction or loss of blood flow signals were detected in 88 cases,and normal blood flow signals were presented in 3 cases.The diagnosis of testicular torsion was confirmed by CDFI in 91 cases before surgery,and 4 cases were diagnosed by medical history and physical examination.The duration from onset to surgery was ＜ 5 h in 10 cases,5-10 h in 25 cases,and ＞ 10 h in 60 cases.All the patients received operation,92 cases were diagnosed as testicular torsion in scrotum and 3 cases as cryptorchidism torsion after surgery.Among 92 cases of testicular torsion in scrotum,twisted testicles were removed in 41 cases and the testicles were retained in 51 cases;while testicles were removed in 1 case of cryptorchidism torsion,and the testicles were fixed in the scrotum in other 2 cases of cryptorchidism.Sixty eight patients were postoperatively followed-up for 1 month to 15 years,the retained necrotic testis atrophy occurred in 1 case,and the testicular size was normal in other 67 cases.In the 68 cases,there was no testicular discomfort and no testicular torsion after operation.Conclusions CDFI is valuable in preoperative diagnosis of testicular tortion.Once the diagnosis is made,it should be operated as soon as possible to retain the testicular function,and the affected testicle should be retained with caution for cryptorchidism torsion.

Objective To analyze and identify the mutations in SGLT2 gene of nine Chinese families with FRG, and determine the renal threshold for glucose excretion (RTG), so as to explore the association of genotype and RTG. Methods All coding regions of SGLT2 gene, including intron exon boundaries, were analyzed using PCR followed by direct sequence analysis. Quantitative test for 24?hour urine glucose and RTG were measured among 9 probands (21 patients) and their family members from 9 pedigrees (total 25 subjects). The differences in renal glucose thresholds between patients with different genotypes (heterozygotes and compound heterozygotes; c.886(-10_-31) del heterozygotes and other heterozygotes) were compared. Results Twelve mutations were identified by SGLT2 gene analysis, including 10 novel ones that were not included in HGMD:c.331T>C, p.W111R;c.374T>C, p.M125T; c.394C>T, p.R132C; c.612G>C, p.Q204H; c.829C>T, p.P277S; c.880G>A, p.D294N;c.1129G>A, p.G377S; c.1194C>A, p.F398L; c.1540C>T, p.P514S; c.1573C>T, p.H525Y. In thisstudy, the mutation c.886(-10_-31)del that is specific to Chinese population accounted for about 28％of the total alleles (5/18). The RTG values of patients with compound heterozygous mutations were much lower than those with simple heterozygous mutations [(1.28 ±0.10) vs (5.14±0.77) mmol/L; P<0.001];and c.886(-10_-31)del heterozygotes had significant lower RTG values than others [(4.43 ± 0.37) vs (5.70 ± 0.51) mmol/L, P<0.001]. Conclusions Ten novel mutations which may be related to FRG are found in this study, and c.886(-10-31)del may be a hot?spot mutation in Chinese patients. Compound heterozygotes had much lower RTG values than simple heterozygotes.