Five minor ferulic acid esters (Ⅰ～Ⅴ) were identified in a petrol extract of Melia azedarach stem bark, one (Ⅰ) of which was isolated with no impurity for the first time. By chemical and spectral methods, their structures were elucidated as hexacosylferulate (Ⅰ); tetracosylferulate (Ⅱ); pentacosylferulate (Ⅲ); heptacosylferulate (Ⅳ) and octacosylferulate (Ⅴ), respectively.

ObjectiveTo evaluate the anti-glaucoma effect of 1％ D-timolol on patients with primary open-angle glaucoma （POAG） and ocular hypertension （OH）．MethodsThirty-six patients with POAG or OH were included．They were divided into two groups randomly for treating with either 1％ D-timolol or 0.5% L-timolol twice daily．The intraocular pressure（IOP） changes and side effects on eyes and cardiovascular system were noticed．Data of visual field and ocular blood flow were compared with the baseline after given the eyedrops for 3 month．ResultsAfter given one drop of the drug，intraocular pressure dropped significantly in both of the two groups．The IOP day curve at 1 month also dropped when compared with the baseline in both groups．There was no significant difference between the two drugs in magitude of hypotensive effect after 3 months．And the higher the IOP was，the more the IOP dropped when using D-timolol．There were no serious side effects noted．No significant change of ocular blood flow was found by color doppler imagine．Mean sensitivity of visual field was raised significantly in D-timolol-treated group，while in L-timolol-treated group it was not changed much．Conclusion1％ D-timolol is a favorable eyedrop. It is effective and safe for the treatment of glaucoma．

Objective To study the effect and possible mechanism of diazoxide on myocardial ischemia/reperfusion (I/R) injury in diabetic rats, and the influence of insulin intervention which aims to maintain blood sugar levels within the normal range on the protective function of cardiomyocytes. Methods 126 health male Sprague-Dawley (SD) rats were intraperitoneally injected with one dose of 60 mg/kg streptozotocin (STZ) to reproduce diabetic model. The diabetic rats were randomly divided into seven groups, with 18 rats in each group. Myocardial I/R model was established by 30 minutes ligation of the left anterior descending branch of the coronary artery, and 120 minutes blood circulation recover. Sham group was only threaded without ligation. Rats in I/R group, diazoxide group (DZ group), and Ottawa vine penicillin (WNT) group were infused intravenously with 2 mL of 0.1% dimethyl sulphoxide (DMSO), DZ (7 mg/kg), and WNT (15 μg/kg), respectively, after 25 minutes of ischemia. Sham group was only injected with 2 mL of 0.1% DMSO. DZ+WNT group was infused with WNT 5 minutes before the injection of DZ. Insulin intervention (RI) group received a continuous insulin infusion to maintain the blood sugar at the level of 4-6 mmol/L. RI+DZ group was infused with DZ after ischemia for 25 minutes based on blood sugar control. Hemodynamic parameters in each group were monitored continuously. The pathological changes of myocardium were observed with hematoxylin and eosin (HE) staining. The expressions of phosphorylated protein kinase B (p-Akt) and phosphorylated glycogen synthase kinase-3β (p-GSK-3β) were determined by Western Blot. Results Compared with sham group, the cardiac functions of the intervention groups were significantly decreased, and severe myocardial injury was observed. Compared with I/R group, the cardiac functions of intervention groups were not obviously improved. However, after insulin intervention by which blood sugar was maintained within normal range, the cardiac function and myocardial injury were further aggravated. Compared with sham group (the expression value of sham group was set as 1), the expressions of p-Akt in other groups including I/R group, DZ group, RI group, and RI+DZ group showed no statistically significant difference (gray value: 1.07±0.09, 1.03±0.07, 1.07±0.07, 1.02±0.08 vs. 1.00, all P > 0.05). However, the expressions of p-Akt were decreased in WNT group and DZ+WNT group as compared with those of sham group and I/R group (gray value: 0.54±0.06, 0.51±0.05 vs. 1.00 and 1.07±0.09, all P < 0.05). The expressions of p-GSK-3βshowed no statistically significant difference in I/R group, DZ group, WNT group, and DZ+WNT group as compared with sham group (gray value: 0.97±0.08, 1.00±0.11, 0.98±0.06, 0.97±0.09 vs. 1.00, all P > 0.05). However, the expression of pGSK-3β was increased in RI group, RI+DZ group as compared with sham group and I/R group (gray value: 1.68±0.08, 1.70±0.05 vs. 1.00 and 0.97±0.08, all P < 0.05), and it was significantly higher in RI+DZ group than that of DZ group (gray value: 1.70±0.05 vs. 1.00±0.11, P < 0.05). Conclusions Diazoxide after myocardial injury could not protect the myocardium from I/R injury in diabetic rats, and did not trigger the activation of the phosphoinositide 3-kinase (PI3K)/Akt signaling pathway. Insulin intervention by which blood sugar was maintaine d within the normal range exacerbates myocardial I/R injury in diabetic rats.

Objective To study the effect of foot motor imagery on lower extremity motor function in hemiplegics after stroke. Methods 72 hemiplegics were randomly divided into two groups: the experiment group and the control group. Normal rehabilitation was performed in the both groups and motor imagery was added in the experiment group. They were assessed with lower extremity Fugl-Meyer Assessment (FMA) and gait analysis. Results The scores of FMA in knee, ankle and lower extremity, gait frequency and speed improved in experiment group more than in controls（P<0.05）. Conclusion Foot motor imagery can improve lower extremity motor function.

Objective To assess the diagnostic value of MR imaging of the wrist synovitis in early rheumatoid arthritis.Methods 60 wrists in 30 patients underwent MR imaging examination,including T1WI,T2WI,STIR and contrast-enhanced fat-suppressed 3D fast-gradient-echo sequence.MRI manifestations were analysed in comparison with clinical and laboratory test data.Results All changes including synovial hyperplasia and pannus formation(30 cases),articular cartilage destruction(21 cases),bone erosion(16 cases),joint effusion(25 cases) and tendon lesion of wrists could be showed clearly by MRI.The signal intensity of the pannus before and after administration of contrast agent and the enhanced curve of the pannus could be used to assess the lesions whether or not activation.Conclusion MRI is helpful in diagnosing early rheumatoid arthritis.

Objective To investigate effects of hyperpolarization-activated cyclic nucleotide-gated cation nonselective channel in the hu-man ureter on the spontaneous contraction of smooth muscles. Methods Four HCN subtypes were detected in human ureteral tissue using reverse transcription polymerase chain reaction,Western blotting and immunohistochemical. ZD7288,the HCN blocker, was used to observe the changes of ureteral muscle contraction amplitude and frequency by applying the ureteral smooth muscle strip test in vitro. Results HCN1-4 isoforms were all identified in human ureter using reverse transcription-polymerase chain reaction and Western blotting. Through the immunohistochemical,HCN channel was found mostly in the urothelium layer and muscular layer of human ureteral wall. ZD7288 significantly decreased the bladder excitation. Conclusion All 4 HCN channel hypotypes exist in the human ureter, and affect the ureteral excitation.

OBJECTIVE:To investigate the effects of saikosaponin-d(SSd) on tissue plasminogen activator(TPA),plasminogen activator inhibitor(PAI),malonaldehyde(MDA) and NO in rats with liver fibrosis induced by dimethylnitrosamine(DMN).METHODS:Eighteen SD healthy rats were randomized to control group(NS ip qd for 4 weeks),model group(10mg? kg-1 DMN ip 3 times per week for 4 weeks) and SSd-treated group(10mg? kg-1 DMN ip 3 times per week + SSd 1.8 mg? kg-1 ip for 28 consecutive days).All rats were killed 1h after the last time of administration,blood samples were taken from abdominal aorta and liver samples were taken for the observation of pathology and detection of indices of TPA,PAI,MDA and NO etc.RESULTS:SSd could lessen the degree of liver fibrosis and improve the fibrinolytic activities of TPA and PAI,meanwhile,it showed clearance effect on MDA and marked protective effect on hepatic cells.There were significant differences between SSd-treated group and the model group.CONCLUSION:SSd exhibited protective function on experimental hepatic fibrosis in rats,which may be attributed to the improving of fibrinolytic activity,eliminating of lipid peroxidation and enhancing of NO level.

AIM: To construct the plasmid vectors containing different regions of human eNOS promoter coupled to a red fluorescent protein reporter gene, which may express in mammalian cells. METHODS: Different regions of human eNOS promoter were subcloned respectively into a red fluorescent protein vector, pDsRed1-1. These recombinant vectors, pDsF1033Red, pDsF494Red and pDsF166Red, were then transfected into NIH3T3 cell lines, followed by the observation under a fluorescent microscope. RESULTS: After identified to be right by double restriction enzyme digestion, PCR and sequencing, the vectors might be effectively expressed in NIH3T3 cells. 95 % of the red fluorescent emitted by a red fluorescent protein dispersed all over the cells, appearing at 48-60 h after transfection, reaching peak at 96-144 h, becoming the strongest in light at 144 h, gradually disappearing after 168 h and remaining little red fluorescent in 21 days. The quantity and intensity in expressions of red fluorescent protein drived by different regions of human eNOS promoter were clearly lower than by a strong promoter, p CMVIE . CONCLUSION: The red fluorescent protein reporter gene vectors containing different regions of human eNOS promoter are successfully constructed and may efficaciously express in mammalian cells, appearing not strong transcriptional activities, which provide practical and feasible tools to study functions of different regions of human eNOS promoter and roles of cis-elements in it. [

OBJECTIVE: To explore the laboratory phenotype and molecular pathogenesis in a Chinese pedigree affected with Hereditary coagulation factor Ⅻ (FⅫ) deficiency. METHODS: A male proband admitted to Ningbo No.2 Hospital on July 17, 2021 due to chronic gastritis and members of his pedigree (7 individuals from three generations) were selected as the study subjects. Prothrombin time (PT), activated partial thromboplastin time (APTT), FⅧ activity (FⅧ: C), FⅨ activity (FⅨ: C), FⅪ activity (FⅪ: C), FⅫ activity (FⅫ: C), and FⅫ antigen (FⅫ: Ag) were determined. All of the exons, exon-intronic boundaries, as well as the 5'- and 3'-untranslated regions of the F12 gene were subjected to Sanger sequencing. Candidate variants were verified by cloning sequencing. The effect of candidate variants on the protein function was analyzed by bioinformatics software. RESULTS: The proband, a 47-year-old male, had significantly prolonged APTT (180.0 s) and decreased FⅫ:C and FⅫ:Ag levels (< 1%). His father, mother, brother and two sons also showed certain degrees of reduction. Genetic testing revealed that the proband has harbored compound heterozygous variants of the F12 gene, namely c.1092_1093insC (p.Lys365Glnfs*69) in exon 10 and c.1792_1796delGTCTA (p.Val579Hisfs*32) in exon 14. His mother and elder son were heterozygous for the c.1092_1093ins variant, whilst his father, brother, and younger son were heterozygous for the c.1792_1796delGTCTA variant. Analysis of the promoter region of exon 1 also showed that the proband and both sons had harbored a 46T/T polymorphism, whilst other family members were 46C/T. Bioinformatic analysis suggested that the p.Val579 is a highly conserved site. Protein model analysis showed that, with the p.Val579Hisfs*32 variant, a benzene ring was added and the hydrogen bond of surrounding amino acids was changed. Based on the guidelines from the American College of Medical Genetics and Genomics, the c.1792_1796delGTCTA was classified as a pathogenic variant (PVS1+PM2_Supporting+PM4). CONCLUSION The c.1092_1093insC (p.Lys365Glnfs*69) and c.1792_1796delGTCTA (p.Val579Hisfs*32) compound heterozygous variants of the F12 gene probably underlay the decreased FXII levels in this pedigree. Above finding has also enriched the mutational spectrum for FⅫ deficiency.
Male ; Humans ; Aged ; Middle Aged ; Pedigree ; East Asian People ; Exons ; Introns ; Family ; Factor XII Deficiency/genetics* ; 3' Untranslated Regions ; Factor XII/genetics*

OBJECTIVE: To explore the genetic characteristics of a child with mosaicism Turner syndrome. METHODS: A child who had presented at Linyi People's Hospital on May 19, 2022 due to short stature was selected as the study subject. The child was subjected to combined chromosomal karyotyping, fluorescence in situ hybridization (FISH), and chromosomal microarray analysis (CMA). RESULTS: The child was found to have a 46,X,i(X)(q10)[94]/45,X[6] karyotype. The result of FISH was nucish(XYpter,XYqter)1[78]/(XYpter)1,(XYqter)3[122]. CMA result for her peripheral blood sample was arr[hg19]Xp22.33p11.1(168551_58526888)×1, and that for her oral mucosal cells was arr[hg19]Xp22.33p11.1(168551_58526888)1-2,Xq11.2q28(63000001_155233098)×2-3. By integrating the above findings, her molecular karyotype was determined as mos 46,X,i(X)(q10)[94]/45,X[6].arr[hg19]Xp22.33p11.1(168551_58526888)×1-2,Xq11.2q28(63000001_155233098)×2-3.nucish(XYpter)1,(XYqter)3[122]/(XYpter,XYqter)1[78], which has indicated mosaicism Turner syndrome. CONCLUSION The 46,X,i(X)(q10)/45,X mosaicism probably underlay the pathogenesis in this child.
Humans ; Child ; Female ; Turner Syndrome/genetics* ; Mosaicism ; In Situ Hybridization, Fluorescence ; Karyotyping ; Karyotype