Objective To study the tissue-specificity of ?-carotene-15,15’-monooxygenase(? CMOOX). Method Semi-quantitative RT-PCR and HPLC were used to study the expression of ?CMOOX mRNA and its activity in different tissues of chicken. Results ?CMOOX mRNA was expressed in tissues including heart, liver, spleen, lung, kidney, duodenum, jejunum, ileum, muscle and testis, not expressed in lung. Furthermore, it was expressed in jejunum with the highest level. ?CMOOX activity was found in every tissue, except lung, and highest in duodenum and jejunum. Conclusion The distribution of ?CMOOX in chicken has tissue specificity.

Objective To better understand the clinical characteristics of Familial Idiopathic Basal Ganglia Calcifi?cation (FIBGC), including at the perspective of hereditary pattern, clinical test results, onset age, clinical heterogeneity and the volume of basal ganglia calcification (VBGC). Method 8 Eight FIBGC families were collected and draw family pedigrees were draw. Analysis of was conducted on the patient's clinical test results, head CT and MRI changes, onset ag?es, relationship of clinical manifestations with VBGC. Results No significant difference was found in serum calcium, alu?minum, arsenic, cobalt, magnesium, phosphorus, iron, parathyroid hormone and calcitonin concentration between the fam?ily members of patients and healthy controls (P>0.05). Family members from 8 FIBGC families including the two with consanguineous marriage manifested autosomal dominant heredity. The severity of , symptomatic s was correlated with VBGCpatients showed the same clinical manifestations in the dyskinesia family. The psychiatric symptoms was not asso? ciated with VBGC whereas patients with dyskinesia had a large VBGC. There was a significant difference in onset age be?tween patients with psychiatric symptoms and those with dyskinesia. P.atients with dyskinesia suffer larger VBGC, and is characterized by Patients with dyskinesia had relatively later onset age (43.95 ± 2.47 y) whereas those with. psychiatric symptoms hadsymptomatic patients with early onset age (31.32±10.16y). The comparison of the onset age (43.954±2.473 vs. 31.319±10.156 y, t=4.438, P=0.001) and VBGC (1.748±0.622 vs. 0.392±0.276 cm3, t=2.518, P=0.028) with symptom?atic patients between dyskinesia and psychogenic families was significant. Conclusions Eight FIBGC families manifested autosomal dominant heredity. Patients with dyskinesia suffer have a larger VBGC and are associated with a, and is char?acterized by relatively later onset age. In contrast, patients with psychiatric symptomspsychogeny is not related withhave a the small VBGC and showedand their age of onset is young. earlier onset age.

BACKGROUND: Infrared spectrum technology has potential and important value of application in diagnosing disease and analyzing pathology, as well as providing references for proper treatment. OBJECTIVE: To analyze the distinctive pathological information in the spectrums of infrared radiation at Neiguan (PC 6) acupoint of patients with coronary atherosclerotic heart disease (CAHD). DESIGN: Case-control study.SETFING: Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine. PARTICIPANTS: The study was conducted from April 2003 to April 2005 in patients with CAHD from Out-Patient Clinic in Longhua Hospital (21 males and 29 females, averagely 58.1 years). Forty-seven healthy volunteers were all from advertisement (25 males and 22 females, averagely 50.6 years). All the subjects were voluntary. METHODS: PHE201, a high-sensitive infrared spectrum detective device (Shanghai Institute of Technical Physics of Chinese Academy of Sciences, China) was applied to detect the spectrums of infrared radiation at Neiguan in 50 patients with CAHD and 47 healthy adults were selected as control for comparison. The scanning range was from 1.5μ m to 16μ m,73 detected wavelength spots were scanned at Neiguan of all the subjects, and the scanning spacing was 0.2μ m. MAIN OUTCOME MEASURES: Intensity of infrared radiation at Neiguan between CAHD patients and normal controls at different wavelengths. RESULTS: The intensities of infrared radiation of CAHD patients were significantly different from that of the healthy volunteers at 32 spots of the right Neiguan as well as at 23 spots of the left Neiguan among the 73 detected wavelength spots(P < 0.05-0.001). At 2-2.5μ m wavelength spots that were related to the energy metabolism, the intensities of infrared radiation of CAHD patients were significantly lower than the healthy volunteers (P < 0.05-0.005). Concerning the difference in intensities of infrared radiation between the left and the right Neiguan, the CAHD patients had more wavelength spots than healthy volunteers did (P < 0.001). CONCLUSION: ①Distinctive pathological information exists in the infrared spectrums at Neiguan in CAHD patients. ②CAHD patients have hypo-activities of energy metabolism and hypo-function ofqi and blood in the area of Neiguan.

Three plasmids (pGenesil-P1, pGenesil-P2, pGenesil-P3) with different p27Kip1-shRNA sequences were designed and synthesized. Their effects on the proliferation of bovine corneal endothelial cells (bCEC) were investigated. Plasmid expressing irrelevant shRNA with a random combination was used as negative control (pGenesil-HK). The recombination of four plamids was confirmed by restrictive enzyme digestion and sequence analysis. The expression of mRNA and protein of p27Kip1 was detected by RT-PCR and Western blotting after stable transfection. The expressions of p27Kip1 mRNA and p27Kip1 protein of pGenesil-P1 group, pGenesil-P2 group and pGenesil-P3 group were all lower than those in the pGenesil-HK group and the blank group (non-transfected group). pGenesil-P3 had the strongest inhibitory effect and was selected for the next steps. The proliferation rates of the pGenesil-P3 group, the pGenesil-HK group and the blank group were assessed by MTT. The influence of shRNA-p27Kip1 on bCEC cell cycle was detected by flow cytometry (FCM). Compared with the control groups, the proliferation rate of the pGenesil-P3 group was increased significantly, and the ratio of S-phase also increased. It is concluded that shRNA-p27Kip1 could down-regulate the expression of p27Kip1 effectively and increase the proliferation of bCEC. RNA interference (RNAi) may be an effective means to promote the proliferation of CEC.
Cell Proliferation ; Cornea/cytology ; Cyclin-Dependent Kinase Inhibitor p27/*metabolism ; Endothelial Cells/*cytology ; Endothelial Cells/metabolism ; Gene Expression Regulation ; Models, Biological ; Plasmids/metabolism ; RNA Interference ; RNA, Messenger/metabolism ; RNA, Small Interfering/metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tetrazolium Salts/pharmacology ; Thiazoles/pharmacology ; Transfection

Objective To knockout and identify the Antigen 43 (Ag43) in the Escherichia Coli JM109 .Methods Mutation group Ⅱ introns RNA protein complexes (RNP) gene sequence was obtained by Sigma Company′s TargeTron Gene Knockout Sys‐tem and Ag43 gene specific designed PCR primers amplification ,then ,to acquired Ag43 specific recombinant RNP plasmid pACD4K‐Ag4 ,this gene sequence was inserted into the plasmid pACD4K‐C of RNA′s expression .Finally ,pEGFP‐Ag43 was trans‐formed into JM109 and inserted the group Ⅱ intron into the Ag43′s locus by IPTG inducing expression .Results The best insertion locus was between 1 812 and 1 913 .Through the agarose electrophoresis gel ,the RNP gene sequence was consistent with the expec‐ted value (350 bp) .The pEGFP‐Ag43 vector was correctly constructed which was proofed by endonuclease Nhe Ⅰ and Hind ⅡI di‐gestion as predicted products (3 646 and 4 029 bp;7 000 and 550 bp ,respectively ) .The PCR and gene sequence results indicated that the group Ⅱ intron was inserted into the locus between 1 812 and 1 913 in the Ag43 gene .Conclusion Successful knockout of the Ag43 in Escherichia Coli JM109 found basis to further study the Ag43′s function and regard the coli as host bacteria of Ag43 chimeric protein recombinant .

OBJECTIVETo establish the new EST-SSR markers for analyzing the genetic variation of different population of Salvia miltiorrhiza.

METHODIt was dealt with ESTs newly downloaded from Genbank and that of acquired from HMPL lab EGassembler software, and then carried out SSR loci search and SSR type analysis by SSRIT software. After that, it was designed the EST-SSR primer pairs for PCR amplification condition optimization.

RESULTAbundant and high coverage of SSR loci distribution were found in S. miltiorrhiza with having one SSR per 5.8 kb ESTs. Among them, the occurrences of different repeat units were mainly the di- (63.0%) and tri- (35.5%). The CT/AG was the most frequent motif in dinucleotide motif type and the GAA/TCC was the most frequent motif in trinucleotide repeats. Out off 36 primer pairs, 29 primer pairs (80.5%) were successfully amplified in all samples of S. miltiorrhiza while the rest failed to give PCR products at various annealing temperature and Mg2+ concentrations. The selected primer pairs also showed the polymorphism in samples from different S. miltiorrhiza populations.

CONCLUSIONThe newly establishment of EST-SSR markers showed high SSR loci coverage and genetic polymorphisms in S. miltiorrhiza population. It could be used for genetic variation analysis.

Objective To investigate the correlation between baseline serum lipid levels and hematoma enlargement in patients with acute intracerebral hemorrhage (ICH). Methods From October 2013 to January 2018, patients with ICH admitted to the Department of Neurosurgery, Heze Municipal Hospital, were enrolled retrospectively. The first CT scan was completed within 6 h after onset, and the second one was completed at 48 h after onset. Hematoma enlargement was defined as an increase >33 % in the volume of hematoma on CT. The demographic and baseline clinical data in the hematoma enlargement group and the non -hematoma enlargement group were compared. Multivariate logistic regression analysis was used to identify the independent risk factors for hematoma enlargement. Results A total of 470 patients with acute ICH were enrolled, including 187 females (39.8%) and 283 males (60.2%), aged 47-81 years. Seventy-nine patients (16.8%) had hematoma enlargement. The proportion of patients with atrial fibrillation and who used warfarin before onset, as well as age, baseline National Institutes of Health Stroke Scale score, baseline hematoma volume, international normalized ratio, prothrombin time, activated partial thromboplastin time, and thrombin time of the hematoma enlargement group were significantly higher than those of the non -hematoma enlargement group ( all P< 0.05 ), while from the onset to the first CT scan time, total cholesterol, triglyceride, low-density lipoprotein cholesterol levels were significantly lower than those in the non- hematoma enlargement group (all P<0.05). Multivariate logistic regression analysis showed that baseline total cholesterol <3.20 mmol/L (odds ratio [ OR] 1.32, 95% confidence interval [ CI] 1.08-1.83; P=0.004), baseline hematoma volume≥30 ml (1.76,95% CI 1.30-2.15; P<0.001), and using anticoagulant before onset ( OR 2.37, 95% CI 1.81-3.02; P<0.001 ) had significantly independent correlation with hematoma enlargement. Conclusion Baseline total cholesterol <3.20 mmol/L, hematoma volume ≥30 ml, and using anticoagulant before onset were the independent risk factors for hematoma enlargement in patients with acute ICH.

Objective:To explore the effects and mechanism of Salvianolic acid B(SalB)on Parkinson's disease(PD).Methods:Forty-eight C57BL/6 male mice were randomly separated into control group(control)withoutdrugs,model group(MPTP)with intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrah-ydropyridine(MPTP),SalB control group with intraperitoneal injection of SalB,and SalB treatment group(MPTP+SalB).Construction of PD mouse model by intraperitoneal injection of MPTP,and treatment with intraperitoneal injection of SalB.Pole climbing test was applied to assess behavior differences.The time of first black stool excretion and water content of feces were measured to evaluate intestinal dysfunctions.The number of tyrosine hydroxylase(TH)positive cells in substantia nigra and the level of Toll like receptor 4(TLR4)in colon were analyzed by immunohistochemistry.The pathological changes of colonic mucosa were observed by HE staining.The levels of calprotectin(CP)and tumor necrosis factor-α(TNF-α)in colon were determined by ELISA.Western blot was used to determine the level of TH in midbrain,the protein level of TH,tight junction protein(ZO-1),and protein level of TLR4/MyD88/NF-κB signaling pathways which express in colon.Results:Com-pared with the Control group,the climbing time,T-turn time and the first black stool excretion time in MPTP group increased while the fecal water content and the number of TH positive cells in substantia nigra were decreased.Accompanied by TLR4 positive cells in colon,pathological injury score of colonic mucosa,levels of CP and TNF-α in colon increased,expression of TH in midbrain and expression of ZO-1 in colon decreased.Expressions of TLR4,MyD88,Nuclear NF-κB p65 and p-NF-κB p65 in colon increased.Com-pared with MPTP group,SalB treatment shortened the climbing time,T-turn time and the first black stool excretion time in SalB treat-ment group,increased the fecal water content and the number of TH positive cells in substantia nigra,lowered TLR4 positive cells in colon,enhanced expression of TH in midbrain and colon,reduced the pathological injury score of colonic mucosa,significantly decreased levels of CP and TNF-α in colon,enhanced expression of ZO-1 in colon,inhibited expressions of TLR4,MyD88,Nuclear NF-κB p65 and p-NF-κB p65 in colon.Conclusion:SalB can protect the nerves and intestines and alleviate the intestinal inflamma-tion of PD mice,which may be related to the inhibition of TLR4/MyD88/NF-κB signal pathway.

OBJECTIVE:To investigate the omeprazole off-label use and provide reference for rational drug use in clinic. METHODS:In retrospective survey,1838 omeprazole prescriptions were selected from outpatient department during Jun. 2014 to May 2017 by random number table. According to the latest drug instruction,whether the off-label use or not was determined. Off-label use was determined by reviewing domestic and foreign guidelines,literatures and Micromedex database. RESULTS:Among 1838 pre-scriptions,there were 1750 prescriptions of off-label drug use (95.21％),involving 13 items and 3 types of off-label drug use. Main type of off-label drug use was over-indication medication,involving 1747 prescriptions (96.47％). There was no statistical significance in the incidence of off-label drug use between digestive department and non-digestive department or among physicians at different levels(P>0.05). Among 13 items of off-label drug use,7 items were supported by evidence-based medicine evidence of domestic and foreign guideline,expert consensus/suggestion or literature reports,etc.;among which 1 item of off-label drug use were included in Micromedex grading system and other 6 items had no evidence-based medicine evidence. CONCLUSIONS:The phenomenon of omeprazole off-label use is widespread in outpatient department of the hospital,and some off-label drug use are sup-ported by evidences. There are differences in the quality of those evidences. It is suggested to standardize the off-label drug use to avoid legal risks and guarantee the safety of drug use for patients.

Three plasmids (pGenesil-P1, pGenesil-P2, pGenesil-P3) with different p27Kipl-shRNA sequences were designed and synthesized. Their effects on the proliferation of bovine corneal endo- thelial cells (bCEC) were investigated. Plasmid expressing irrelevant shRNA with a random combi- nation was used as negative control (pGenesil-HK). The recombination of four plamids was con- firmed by restrictive enzyme digestion and sequence analysis. The expression of mRNA and protein of p27Kipl was detected by RT-PCR and Western blotting after stable transfection. The expressions of p27Kipl mRNA and p27Kipl protein of pGenesil-P1 group, pGenesil-P2 group and pGenesil-P3 group were all lower than those in the pGenesil-HK group and the blank group (non-transfected group), pGenesil-P3 had the strongest inhibitory effect and was selected for the next steps. The pro- liferation rates of the pGenesil-P3 group, the pGenesii-HK group and the blank group were assessed by MTT. The influence of shRNA-p27Kipl on bCEC cell cycle was detected by flow cytometry (FCM). Compared with the control groups, the proliferation rate of the pGenesiI-P3 group was increased significantly, and the ratio of S-phase also increased. It is concluded that shRNA-p27Kipl could down-regulate the expression of p27Kipl effectively and increase the proliferation of bCEC. RNA interference (RNAi) may be an effective means to promote the proliferation of CEC.