2.Effects of overexpression of human vascular endothelial growth factor 165 and human tissue inhibitor of metalloproteinase 1 on rats with myocardial infarction
Yanqiu SONG ; Yongmin MAO ; Hua GENG ; Ying ZHANG ; Ying SHI ; Min REN ; Meilin XU ; Zhigang GUO
Chinese Journal of Emergency Medicine 2021;30(3):293-300
Objective:To investigate the effects of recombinant adenovirus with human vascular endothelial growth factor 165 (Ad-hVEGF 165) and recombinant adenovirus with human tissue inhibitor of metalloproteinase 1 (Ad-hTIMP-1) on rats with myocardial infarction (MI) and its mechanism. Methods:A total of 30 healthy 8-week-old male Wistar rats were randomly divided into 5 groups: sham-operated group (sham), virus control group (Ad-Track), Ad-hVEGF 165 group, Ad-hTIMP-1 group and Ad-hVEGF 165+Ad-hTIMP-1 group (hVEGF 165+hTIMP-1) ( n=6 per group). Except the sham group, all rats were ligated the left anterior descending coronary artery to induce MI model with ST-segment elevation and Q waves or T-wave inversion on electrocardiogram and local myocardial whitening. The corresponding recombinant adenovirus comprising 100 μL (1×10 10 VP/100 μL) combined with NaCl solution was injected into the myocardial infarction area at four points respectively. The sham group received no treatment. After 4 weeks, all rats were sacrificed after echocardiography was completed and heart tissues were collected. The expression of hVEGF 165 and hTIMP-1 were detected by immunohistochemistry. The mRNA expression of apoptosis-related factors were detected by real-time PCR. The protein expression of apoptosis-related factors were detected by immunohistochemistry. Differences between groups were determined by One-way analysis of variance. Multiple comparisons between groups were performed using the least significant difference t-test. Results:(1) Both heart rate (HR) (480.83±24.09) beats/min, left ventricular end-diastolic dimension (LVEDD) (6.88±0.44) mm and left ventricular end-systolic dimension (LVESD) (4.85±0.42) mm were increased in the Ad-Track group than those in the sham group (433.16±17.86) beats/min, (6.20±0.45) mm, (4.06±0.70) mm (all P<0.05), and left ventricular ejection fraction (LVEF) (62.70±3.17) % and left ventricular fractional shortening (LVFS) (29.52±1.88) % were significantly decreased in the Ad-Track group than those in the sham group (72.78±5.44)%, (29.52±1.88) % (both P<0.01). Compared with the Ad-Track group, LVEF (71.50±6.23) % and LVFS (36.17±5.27) % in the hVEGF 165-hTIMP-1 group were significantly increased (both P<0.01), and LVEDD (6.22±0.39) mm and LVESD (4.13±0.23) mm were decreased (both P<0.05). LVEF and LVFS in the hVEGF 165-hTIMP-1 group were increased significantly than those in the Ad-hVEGF 165 group (64.65±4.00) %, (30.95±2.57) % (both P<0.05). The mRNA expression of BCL2-associated X protein (Bax), cysteine aspartate specific proteinase 3 (Caspase-3) and BCL-xL/BCL-2-associated death promoter (Bad) in the hVEGF 165-hTIMP-1 group were decreased than those in the Ad-Track group ( P<0.01 or P<0.05), and B-cell lymphoma/leukemia-2 (Bcl-2) in the hVEGF 165-hTIMP-1 group were increased than those in the Ad-Track group ( P<0.01). The mRNA expression levels of Bax and Caspase-3 in the hVEGF 165-hTIMP-1 group were decreased than those in the Ad-hVEGF 165 group (both P<0.05). There was no statistically difference in the mRNA expression of Bax, Caspase-3, Bad, and Bcl-2 between the hVEGF 165-hTIMP-1 group and the sham group (all P>0.05). The protein expression of Bax and Caspase-3 in the hVEGF 165-hTIMP-1 group were significantly decreased than those in the Ad-hVEGF 165 group, the Ad-hTIMP-1 group and the Ad-Track group (all P<0.01), and the protein expression of Bcl-2 in the hVEGF 165-hTIMP-1 group was increased than those in the Ad-hVEGF 165 group, the Ad-hTIMP-1 group and the Ad-Track group (all P<0.05). There were no statistically differences in the protein expression of Bax, Caspase-3 and Bcl-2 between the hVEGF 165-hTIMP-1 group and the sham group (all P>0.05). Conclusions:Ad-hVEGF 165 and Ad-hTIMP-1 can improve cardiac contractile function of MI rats and the beneficial effects are largely attributable to inhibiting myocyte apoptosis. The combination of hVEGF 165 and hTIMP-1 may have a synergistic effect on MI.
3.Effect and Mechanism of Extracts of Nidus Collocaliae on Resisting Avian Influenza A H5N1 Virus
Jieru LIN ; Ying ZHANG ; Weiping DAI ; Shuwen LIU ; Geng LI ; Hua ZHOU ; Xiaoping LAI
Journal of Guangzhou University of Traditional Chinese Medicine 2016;33(5):710-715
Objective To observe the inhibitory effect of the extracts of Nidus Collocaliae on avian influenza A H5N1 virus in vitro. Methods Nidus Collocaliae water extract, artificial gastric juice digestion products of Nidus Collocaliae water extract, and artifitial intestinal juice digestion products of Nidus Collocaliae water extract were prepared for the experimental study. 293T cells transfection in vitro was carried out. The effects of 3 kinds of Nidus Collocaliae extracts on H5N1 pseudovirus and VSV-G pseudovirus were determined by luciferase detection kit. Blood clotting response to erythrocyte hemagglutinin subtypes H5, H7, H9 antigens and the inhibitory effects of 3 kinds of Nidus Collocaliae extracts were observed. The effects of 3 kinds of Nidus Collocaliae extracts on neuraminidase activity were determined by neuraminidase inhibitor screen kit. Results The 3 kinds of Nidus Collocaliae extracts had inhibitory effects on H5N1 avian influenza pseudovirus, the effects being enhanced with the increase of the concentrations of Nidus Collocaliae extracts. Of the 3 extracts, artificial intestinal digestion products had the strongest inhibitory effect, while Nidus Collocaliae water extract had the weakest effect. However, Nidus Collocaliae extracts had no obvious effect on VSV-G pseudovirus. The concentration of H5, H7 antigen for positive blood clotting response was 1 ∶ 128, and that of H9 antigen was 1 ∶ 256. The 3 kinds of Nidus Collocaliae extracts at certain concentrations could inhibit blood clotting response to H5, H7, H9 antigen, but had no obvious effect on neuraminidase. Conclusion The anti-H5N1 virus effect of Nidus Collocaliae extracts has been achieved probably through resisting hemagglutinin.
4.Influence of naturaldecay of fibrousroots on growth and content of active compositions of Salvia miltiorrhiza f. alba.
Wei LIU ; Ying-ying WEI ; Peng SUN ; Jie ZHOU ; Xiao WANG ; Yan-ling GENG ; Jian-hua LIU ; Feng-sheng LI
China Journal of Chinese Materia Medica 2015;40(13):2548-2552
The change of yield and contents. of active compositions were studied while the fibrous roots were decayed naturally. HPLC method was used to detect the contents of active composition. The results show that fibrousroots could decrease the production of plant by 38.60% (20 g) and 30.99% (40 g), respectively. Treatment 1 could increase the contents of dihydrotanshinone and cryptotanshinone of Salvia miltiorrhiza f. alba by 26.08% and 22.64%, respectively. Compared with the comparison, treatment 2 decreased the contents of ihydrotanshinone, cryptotanshinone, tanshinone I and tanshinone II(A) of S. miltiorrhiza f. alba by 60.87%, 79.24%, 84.61% and 88.99%, respectively. Meanwhile, the total contents of the liposoluble constituents reduced by 86.27%. The different concentration of fibrousroots could increase the content of salvianolic acid B by 4.98% (20 g) and 23.64% (40 g), respectively. Meanwhile, the content of rosemary acid was increased by 4.98% (20 g) and 23.64% (40 g), respectively. The content of water-soluble constituents positively correlated to the mount of added fibrousroots, and the change was significantly. The result indicted that the decay of fibrousroots has a significant impact on the growth and the content of the active composition of S. miltiorrhiza f. alba under the condition of continuous cropping. Fibrousroots could decrease the content of biomass and liposoluble constituents significantly, which maybe one of the main factors to S. miltiorrhiza f. alba continuous cropping obstacle formation.
Benzofurans
;
analysis
;
Biomass
;
Chromatography, High Pressure Liquid
;
Diterpenes, Abietane
;
analysis
;
Plant Roots
;
metabolism
;
Salvia miltiorrhiza
;
chemistry
;
growth & development
5.Effects of coriaria lactone-activated astrocyte conditioned medium on the expressions of glutamate and GluR2 in the rat brain
Shu-Hua ZHANG ; Chang-Geng ZHU ; Qing-Ying LIU ; Ying WEI ; Wei WANG
Chinese Journal of Neuromedicine 2008;7(6):549-553
Objective To explore the effects of coriafia lactone (CL)-activated astrocytes (Ast) conditioned medium (ACM) on the expressions of glutamate (Glu) and GluR2 in the brain of rat. Methods Asts of hippocampus were cultured according to the McCarthy and DeVellis's method, and then the ACM was collected. Forty-eight male adult Sprague-Dawley (SD) rats were randomly divided into the control group (n=16) and the CL group (n=32). Rats in the control group were administered 10 μL ACM I. C. V., which was not added any stimulating substance. Rats of the CL group were injected I. C. V. 10 μL CL-activated ACM. The rats in both groups were subdivided into post-injection 2,4,8,12h subgroups, 4 in each subgroup in the control group and 8 in each subgroup in the CL group. The behaviors of the rats were observed and the expressions of Glu and GluR2 in the cerebral cortex and hippocampus were detected with immunohistochemistry and immunofluorescence. The content of GluR2 was tested with Western blot. Results The rats injected with CL-activated ACM showed seizure activities, whereas the rats of the control group showed no seizure activities. The expression of Glu in cerebral cortex and hippocampus in the brains injected with CL-activated ACM was increased compared with the control group at 4h (P<0.05), but the expression of GluR2 was attenuated compared with the control group at 4h(P<0.05). The results of GluR2 in the cerebral cortex and hippocampus detected with Western blot were different significantly with control group (P<0.05). Conclusion CL-activated ACM can enhance the expression of Glu and reduce the expression of GluR2 in the brain of rat, resulting in the activation of AMPA pathway and the Ca2+ influx, and then induce seizure activities.
6.Observation on theraputic effect of five-needle-in-nape acupuncture for treatment of poststroke pesudobulbar paralysis dysphagia.
Min LI ; Jian-hua SUN ; Jun-wei LU ; Lan-ying LIU ; Hao GENG
Chinese Acupuncture & Moxibustion 2009;29(11):873-875
OBJECTIVETo compare the therapeutic effect of five-needle-in-nape acupuncture treatment and the routine acupuncture treatment for poststroke pesudobulbar paralysis dysphagia.
METHODSSixty cases were randomly divided into a five-needle-in-nape (FNN) group and a routine acupuncture (RA) group (n=30 in each group). Ya-men (GV 15), Tianzhu (BL10), Zhiqiang acupoint (Extra) were needled and the training of swallowing was practiced when needling in FNN group. Lianquan (CV 23), Tongli (HT 5), Zhaohai (KI 6) were needled in RA group. The course of acupuncture treatment consisted of needling once time every day, six times a week for two weeks. The therapeutic effects were evaluated after two courses of acupuncture treatment.
RESULTSThe effective rate was 93.3% in FNN group, better than that of 80.0% in RA group (P < 0.05).
CONCLUSIONThe therapeutic effect in FNN group was better than that of RA group for poststroke pesudobulbar paralysis dysphagia.
Acupuncture Points ; Acupuncture Therapy ; Aged ; Bulbar Palsy, Progressive ; etiology ; physiopathology ; therapy ; Deglutition ; Deglutition Disorders ; etiology ; physiopathology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Stroke ; complications ; Treatment Outcome
7.Comparison of self-microemulsifying drug delivery system versus solid dispersion technology used in the improvement of dissolution rate and bioavailability of vinpocetine.
Ying CHEN ; Gao LI ; Jian-Geng HUANG ; Rui-Hua WANG ; Hong LIU ; Ren TANG
Acta Pharmaceutica Sinica 2009;44(6):658-666
The objective of this study is to compare the differences between self-microemulsifying drug delivery system (SMEDDS) and solid dispersion (SD) technology used to improve the dissolution rate and bioavailability of vinpocetine (VIP). The formulation of VIP-SMEDDS was composed of Labrafac, oleic acid, Cremophor EL, Transcutol P, and gum acacia which was used as solid absorbent. VIP-SD was prepared using poloxamer F68 as the carrier. In the solubility test, the solubility of VIP in SMEDDS was 17.3 times as much as that in SD. In the dissolution test, SMEDDS had shown better enhancement and stability in dissolving VIP than SD. When compared to VIP crude powder, the bioavailability of VIP in SMEDDS (VIP-SMEDDS) was 1.89-fold higher, and was less affected by food intake. However, the bioavailability of VIP in SD (VIP-SD) was bioequivalent to that of VIP crude powder. The tissue uptake of VIP-SMEDDS in Peyer's patches, intestine and liver after administration for 2 hours was more favorable than that of VIP-SD, which was 3.7 times higher in Peyer's patches, 2.2 times higher in intestine and 1.5 times higher in liver. In Caco-2 tests, the apparent permeability (P(app)) of VIP-SMEDDS was 2.65 times of that of VIP-SD. The width of the cell tight junctions of Caco-2 cell monolayer treated with VIP-SMEDDS were 9.6-fold wider, but there was no significant change after treatment with VIP-SD, when compared to the blank control. In conclusion, SMEDDS was more efficient than the traditional SD technology in increasing solubility, dissolution, intestinal permeability, lymphatic absorption and bioavailability of the insoluble drugs such as VIP, which is less affected by food intake.
Animals
;
Biological Availability
;
Caco-2 Cells
;
Dosage Forms
;
Drug Delivery Systems
;
Emulsions
;
chemistry
;
pharmacokinetics
;
Humans
;
Male
;
Rats
;
Rats, Sprague-Dawley
;
Solubility
;
Vinca Alkaloids
;
chemistry
;
pharmacokinetics
8.Association Study of Polymorphisms in Neuronal Nicotinic Acetylcholine Receptor Subunit Genes With Schizophrenia in the Han Chinese Population
Yuan-yuan LI ; Rui-jie GENG ; Shun-ying YU ; Guan-jun LI ; Zhou-ye WANG ; Hua-fang LI
Psychiatry Investigation 2021;18(10):943-948
Objective:
To investigate the relation between nicotinic acetylcholine receptor subunit (nAChR) genes and schizophrenia, and the relation between tag single nucleotide polymorphism (rs1317286, rs1044396, rs6494212, rs16969968, and rs684513) and schizophrenia in Han Chinese people.
Methods:
The protein-protein interaction (PPI) network among nAChR protein and 350 proteins encoded by schizophrenia-related susceptibility genes was constructed through the String database to explore whether nAChR genes were associated with schizophrenia in these known databases. Then, five single nucleotide polymorphisms (SNPs) of CHRNA3 (rs1317286), CHRNA4 (rs1044396), CHRNA7 (rs6494212), and CHRNA5 (rs16969968, rs684513) were analyzed in a sample of 1,035 schizophrenic patients and 816 healthy controls. The interaction between the markers was analyzed using multifactor dimensionality reduction (MDR) software. Power analysis was performed using the Quanto program.
Results:
There are no significant differences in genotype or allele distribution were identified between the patients and controls (p>0.05). The haplotypes constructed by four markers rs1317286, rs6494212, rs16969968, and rs684513 were not associated with schizophrenia either. However, a significant association between models made of rs1317286, rs1044396, rs6494212, and rs684513 and schizophrenia was revealed in interaction analysis (p<0.05).
Conclusion
The nAChR protein may have effects on the development of schizophrenia through the interaction with proteins encoded by schizophrenia-related susceptibility genes, but no relation was found between selected polymorphisms and schizophrenia in the collected Han Chinese people. However, interaction analysis suggested four-SNP model has an important effect on schizophrenia.
9.Examination of anti-HBx in sera from patients of chronic hepatitis B,liver cirrhosis and hepatocellular carcinoma and its clinical significance
Hang ZHANG ; Lian-Ying WU ; Shu-Ye LIU ; Xin-Hua SHAO ; Zong-Wei YANG ; Xiu-Qin ZHANG ; Xue-Li GENG ; Li-Hong YE ; Xiao-Dong ZHANG
Chinese Journal of Laboratory Medicine 2001;0(03):-
Objective To establish a method of detecting hepatitis B virus x antigen (HBxAg) and antibody to HBxAg (anti-HBx) and to demonstrate its clinical significance of HBxAg and anti-HBx in sera from patients of chronic hepatitis B (CHB),liver cirrhosis (LC) and hepatocellular carcinoma (HCC). Methods Full length HBx gene was cloned into pET30a(+),a prokaryotic expression vector,named pET30a-X.It was transformed into Escherichia coli BL21 (DE3),followed the fusion protein of HBx-His was induced by IPTG.The purified fusion protein was used to immunize rabbit as an antigen to generate polyclonal antibody to HBx protein.The method of enzyme-linked immunosorbent assay (ELISA) was established by using purified fusion protein and generated antibody,which was used to detect HBxAg and anti-HBx in sera from patients of CHB,LC,HCC and normal healthy people.Results The positive rates of HBxAg/anti-HBx were 8.7%/10.4% for CHB,17.9%/40.6% for LC,and 9.8%/34.4% for HCC, respectively.In statistics,the positive rates of anti-HBx in LC and HCC were higher than that in CHB (P
10.Change of cytochrome c in postconditioning attenuating ischemia-reperfusion-induced mucosal apoptosis in rat intestine.
Wei-Wei CHU ; Lei NIE ; Xin-Ying HE ; Ai-Li YAN ; Yi ZHOU ; Geng-Li WU ; Dian-Hua WANG
Acta Physiologica Sinica 2010;62(2):143-148
The present study aimed to investigate the change of cytochrome c in postconditioning-attenuated ischemia-reperfusion (I/R)-induced mucosal apoptosis in rat intestine compared with ischemic preconditioning (IPC). Using rat model of intestine I/R injury, male Sprague-Dawley rats weighing 220-250 g were divided into 4 groups which were Sham operation group, I/R group, IPC group and ischemic postconditioning (IPOST) group. In these groups, I/R procedure was performed by the occlusion of the superior mesenteric artery (SMA) for 45 min followed by reperfusion for 1 h. In Sham group, there was no intervention. In IPC group, SMA was occluded for 5 min and reperfused for 5 min, for two cycles, before the prolonged occlusion. In IPOST group, three cycles of 30-s reperfusion and 30-s reocclusion were preceded at the start of reperfusion. After the reperfusion, the small intestines were sampled for experimental detection. Intestinal mucosal mitochondrial membrane potential was detected by confocal laser scanning microscopy. Expressions of cytochrome c and caspase-3 proteins were detected using Western-blot method. The apoptosis of intestinal mucosal cells was determined with agarose gel electrophoresis and deoxynucleotidyl transferase mediated dUTP-biotin nick-end labeling (TUNEL) technique. Compared with I/R group, the mitochondrial membrane potentials and the expressions of cytochrome c protein were significantly increased, while the expressions of caspase-3 and the apoptotic rates were decreased in IPOST and IPC groups (P<0.05). There were no significant differences between IPOST and IPC groups (P>0.05). These data provide substantial evidence that IPOST attenuates I/R-induced mucosal apoptosis by reducing the release of cytochrome c from mitochondria in the rat small intestine.
Animals
;
Apoptosis
;
physiology
;
Cytochromes c
;
metabolism
;
Intestinal Mucosa
;
metabolism
;
pathology
;
Intestines
;
blood supply
;
Ischemic Postconditioning
;
methods
;
Male
;
Membrane Potential, Mitochondrial
;
Mitochondria
;
metabolism
;
Rats
;
Rats, Sprague-Dawley
;
Reperfusion Injury
;
prevention & control