1.The effect of Dimethyl sulfoxide on facilitating the absorption of hepatitis B virus in HepG 2 cells
Linglong REN ; Yongjian GUO ; Yulan LUO
International Journal of Laboratory Medicine 2015;(23):3444-3446
Objective To preliminarily explore the early process of hepatitis B virus (HBV) infection in HepG2 cells induced by dimethyl sulfoxide(DMSO) ,and provide cytological bases for mechanism study of HBV infection in vitro .Methods HepG cells were divided into the DMSO inducing group and control group ,and were cultured 4 days by DMEM containing 1 .5% DMSO and normal DMEM respectively ;changes of cellular morphology were observed .In addition ,selected ECV304 cells as the negative con‐trol group and treated with DMSO .Cells in the three groups were incubated 2 hours with HBV positive serum after culturing 24 hours ,then trypsin digestive solution ,HepG2 cells and ECV304 cells were collected and polymerase chain reaction (PCR) was used for the determination of HBV DNA .Simultaneously ,the blank control group was set ,and the position of HBsAg on HepG2 cells were detected by using indirect immunofluorescence .Results HepG2 cell volume in the DMSO inducing group was obviously in‐creased .HBV DNA was found in HepG2 cells both in DMSO inducing group and control group ,and DMSO inducing group ex‐pressed much stronger .The results of IIF shown that green fluorescent signals of cell membrane and cytoplasm of HepG 2 cells in the DMSO inducing group were increased obviously ,while the results of HBV DNA and IIF both were negative in the negative con‐trol group .Conclusion DMSO could facilitate adsorption of HBsAg in some extent ,which are benefit for completing the process of early infection .
2.Effects of microRNA-490-5p on visceral sensitivity in rat with intestinal dysfunction
Hesheng LUO ; Haixia REN ; Facan ZHANG ; Liexin LIANG ; Guo ZHANG
Chinese Journal of Digestion 2017;37(4):254-259
Objective To investigate the role of microRNA-490-5p (miR-490-5p) in the regulation of visceral sensitivity in rats with intestinal dysfunction.Methods The lentivirus vector system was used to construct the rno-miRNA-490-5p lentivirus expression vector.The rats were divided into normal group,diarrhea-predominant irritable bowel syndrome (IBS-D) group,lentivirus empty vector group and the siRNA silent group and the latter three groups were model groups.The efficiency of siRNA was measured by real-time polymerase chain reaction (PCR).The rats were gavaged with 10% India ink,and then the time of first black stool,water content of feces and threshold of expansion capacity caused abdominal elevation or back arching were calculated.The visceral sensitivity of rats after miRNA-490-5p silenced was evaluated with abdominal withdrawal reflex (AWR) score by stimulating with different intensities of colonic dilatation.The abdominal electrical activity of rats stimulated by colonic distension was measured by BL-420F biological and functional experimental system.The change of the tension of rats isolated colon intestinal stimulated with acetylcholine chloride was also detected by BL-420F biological and functional experimental system.T test was used to compare the differences between the model groups and the normal group.One way analysis of variance was performed for multi-group comparison after miRNA-490-5p interfered.For comparison between two groups among multiple groups,least significant difference (LSD) method was used when the variance was equal,and Games-Howell method was used when the variance was unequal.Results The gastrointestinal propulsion time and the threshold of expansion capacity caused abdominal elevation or back arching of model groups were both lower than those of the normal group ((8.54±4.07) hvs (12.33±2.23) h,(0.56±0.08) mL vs (0.84±0.09) mL),and the differences were statistically significant (t =2.62 and 6.37,both P < 0.05).After distension with 0.8 mL and 1.2 mL sodium chloride solution,the AWR scores of model groups were significantly higher than those of the normal group (3.20±0.56 vs 1.20±0.45,3.73±0.46 vs 2.60±0.55),and the differences were statistically significant (t=7.20 and 4.58,both P<0.01).There was no significant difference in AWR score between the model groups and the normal group when distended with 1.6 mL sodium chloride solution (3.93 ±0.26 vs 3.80 ± 0.45) (P>0.05).After miRNA-490-5p silenced,gastrointestinal propulsion time of normal group,IBS-D group,lentivirus empty vector group and the siRNA silent group was (11.12±1.01) h,(6.23±3.17) h,(6.09 ± 2.26) h and (12.36±1.97) h,and the differences among four groups were statistically significant (F=10.55,P<0.01).The abdominal electrical activity of normal group,IBS-D group,lentivirus empty vector group and the siRNA silent group distension stimulated with 0.8 mL and 1.2 mL sodium chloride solution was (64.91 ± 10.50),(101.79 ±11.73),(80.49±1.27),(66.92±3.24) μV,and (105.09±52.40),(131.71± 16.74),(111.00±6.41) and (95.49± 4.2) μV,and the differences among four groups were statistically significant (F=16.82 and 9.14,both P<0.05).There was no significant difference in abdominal electrical activity amplitude between silenced group and normal group ((66.92±3.24) μV vs (64.91±10.49) μV and (95.49±4.22) μV vs (105.09±2.40) μV) (all P> 0.05).After distension with 1.6 mL sodium chloride solution,the abdominal electrical activity amplitudeof silenced group was lower than the other groups,and the differences were statistically significant (F=11.09,P<0.01).After adding 1∶1 000 acetylcholine chloride added,the tension of colon of normal group,IBS-D group,lentivirus empty vector group and the siRNA silent group increased by 0.71 ± 0.21,0.81±0.06,0.88±0.21 and 0.43±0.07,however there was no significant difference among the four groups (F=2.57,P =0.100).Conclusions Visceral hypersensitivity existed in rats with intestinal dysfunction.miRNA-490-5p may be involved in the regulation of visceral sensitivity.
3.Achilles tendon extension and tibialis transfer combined with tendon transposition for cerebral palsy-induced talipes equinovarus:ankle recovery and foot function evaluation
Jingquan GUO ; Yi LUO ; Yu GAO ; Shangli REN ; Zilei ZHENG
Chinese Journal of Tissue Engineering Research 2015;(2):241-245
BACKGROUND:It has a variety of treatments for equinovarus in spastic cerebral palsy, including physical therapy, orthotic therapy, systemic anti-spasticity drug therapy, local intramuscular injection of botulinum toxin type A and a variety of surgical operations. These treatments aim at different severity of deformities and different age in children patients, and each has its own merits. OBJECTIVE:To investigate the therapeutic effect of Achiles tendon “Z”-type extension, tibialis anterior and posterior transfer combined with tendon transposition to treat equinovarus in spastic cerebral palsy. METHODS: Twenty-two children with spastic cerebral palsy combined with equinovarus were colected from 2012 to 2014, and then these children were subject to Achiles tendon “Z”-type extension, tibialis anterior and posterior transfer combined with tendon transposition. Postoperative plaster external fixation was done for 6 weeks, and then rehabilitation training was given. RESULTS AND CONCLUSION:Al the 22 patients were folowed for an average of 1-2 years, and then were evaluated according to the degree of patient’s satisfaction and the degree of foot deformity. Satisfactory results were obtained from al the patients. Achiles tendon “Z”-type extension, tibialis anterior and posterior transfer combined with tendon transposition to treat spastic cerebral palsy combined with equinovarus is a good method, and it is characterized as good appearance of the ankle and favorable foot function.
4.Comparison of patient-controlled analgesia and epidural morphine following gynecological surgery
Yongfang JIN ; Yuguang HUANG ; Xiangyang GUO ; Zhumin GE ; Hongzhi REN ; Tiehu YE ; Ailun LUO
Chinese Journal of Anesthesiology 1996;0(09):-
Objective: To compare the efficacy and side effects of patient-controlled intravenous morphine with epidural single bolus morphine in postoperative pain relief. Method: Sixty patients undergoing gynecological procedures under epidural anesthesia were randomly assigned to epidural morphine(EPI)group or patient-controlled intravenous analgesia (PCIA) group. In the EPI group,2 mg of morphine was injected into epidural space at the end of operation. In PCIA group, 1 mg of morphine as a demand dose would be injected intravenously by the patient through a patientcontrolled analgesic delivery system until the pain relieved. The patients were followed up at 4, 8, 12, 24 h after operation,and the degree of pain,sedation, nausea and vomiting were assessed. Result: The total dosage of morphine was higher in the PCIA group(19.08?5.0 mg)than that in the EPI group(2mg,P
5.Association of apolipoprotein E gene polymorphism with postoperative delirium
Gang TAN ; Xiangyang GUO ; Ailun LUO ; Yuguang HUANG ; Jianqing XU ; Hongzhi REN ; Tiehu YE
Basic & Clinical Medicine 2006;0(12):-
0.05) in the presence of ApoE ?4 allele.In all four ApoE ?4/4 homozygote patients,one female patient presented a transient delirium status three days before surgery,another male patient presented serious fluctuated delirium symptoms from the second to 17th days after operation.Conclusion The presence of ApoE ?4 allele seems not a predictator of postoperative delirium.ApoE ?4/4 homozygote patients may be more indulgent to delirium than others.
6.Association of apolipoprotein E gene polymorphism with postoperative delirium
Gang TAN ; Xiangyang GUO ; Ailun LUO ; Yuguang HUANG ; Jianqing XU ; Hongzhi REN ; Tiehu YE
Basic & Clinical Medicine 2009;29(12):1301-1304
Objective To investigate whether apolipoprotein E (ApoE) genotypes is associated with postoperative delirium in aged noncardiac surgical patients. Methods Two hundreds and twelve inpatients over 65y, undergoing selective noncardiac surgeries were enrolled in the study. The patients were frequently interviewed and evaluated prospectively for delirium with the Confusion Assessment Method (CAM) during the first three postoperative days. APOE genotype was determined using multiplex amplification refractory mutation system pelymerase chain reaction (multi-ARMS PCR) technique. Results Delirium occurred in 45 patients during the first three postoperative days. Of the 212 patients, 18 (8.5%) possessed one or two ApoE 84 allele. There was no significant difference between delirious patients and non-delirious patients(6.7% : 9.0%, P >0.05) in the presence of ApoE ε4 allele. In all four ApoE ε4/4 homozygote patients, one female patient presented a transient delirium status three days be-fore surgery, another male patient presented serious fluctuated delirium symptoms from the second to 17th days after operation. Conclusion The presence of ApoE ε4 allele seems not a predictator of postoperative delirium. ApoE ε4/4 homozygote patients may be more indulgent to delirium than others.
7.Study on inhibitory effects and mechanism of lipophilic components in Salvia miltiorrhiza on angiogenesis in vitro.
Xue-Mei FAN ; Gui-Xiang REN ; Qiong-Lin LIANG ; Yi-Ming WANG ; Guo-An LUO
China Journal of Chinese Materia Medica 2014;39(4):744-747
In this study, the human umbilical vein endothelial cell model was used to study the regulating effect of lipophilic components in Salvia miltiorrhiza on angiogenesis, and explore its possible mechanism. The cell model was established to determine the effect of lipophilic components in S. miltiorrhiza on the proliferative activity and migration capacity of endothelial cells. Then the realtime fluorescence quantification PCR technology was applied to detect the changes in the gene expressions of angiogenesis-related cytokines VEGF-A, VEGF-C and MMP-9. The results showed that 5 mg x L(-1) lipophilic components in S. miltiorrhiza could inhibit the proliferation and migration of endothelial cells, and reduce the expression of VEGF-A and MMP-9 genes. It indicated that lipophilic components in S. miltiorrhiza may inhibit the proliferation and migration of endothelial cells by inhibiting the expression of VEGF-A and MMP-9 genes, so as to show the inhibitory effect on angiogenesis.
Angiogenesis Inhibitors
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chemistry
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pharmacology
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Cell Movement
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drug effects
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Cell Proliferation
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drug effects
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Drugs, Chinese Herbal
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chemistry
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pharmacology
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Human Umbilical Vein Endothelial Cells
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cytology
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drug effects
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metabolism
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Humans
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Matrix Metalloproteinase 9
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genetics
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metabolism
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Salvia miltiorrhiza
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chemistry
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Vascular Endothelial Growth Factor A
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genetics
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metabolism
8.Rapid screening of 28 alkaloids in food poisoning samples by liquid chromatography-tandem mass spectrometry
ZHAO Ling-guo ; LUO Lan ; YIN Zhen-yi ; REN Yan ; LEI Lei ; MA Zhi-feng
China Tropical Medicine 2023;23(3):260-
Abstract: Objective To investigate a poisoning incident caused by eating eight treasure congee, and establish liquid chromatography (LC)-mass spectrometry (MS)/MS screening method of 28 alkaloids to provide references for disposal of similar poisoning incidents. Methods LC-MS/MS was used for screening 28 alkaloids in the urine, eight treasure congee and food raw material, and the detected alkaloids were quantified. Samples were extracted with 0.4% formic acid aqueous solution and separated by a Acquity UPLC BEH C18 column (1.7 μm, 100 × 2.1 mm). Acetonitrile-0.2% formic acid aqueous solution was used as the mobile phase and gradient elution was adopted. The ionization mode was electrospray positive ionization mode, and the detection method was multi-reaction monitoring (MRM). Analytes were quantified with the external standard method. Results In the concentration range of 0-100 ng/mL, the linear correlation coefficient r were greater than 0.999 for 28 alkaloids. The recovery of 28 alkaloids in urine sample ranged from 63.0% to 105.0%, and the relative standard deviations (RSDs) were between 5.8% and 8.6%. The recovery of 28 alkaloids in eight treasure congee sample ranged from 72.0% to 109.0%, and the RSDs were between 6.3% and 9.7%. The recovery of 28 alkaloids in semen sesami nigrum sample ranged from 60.0% to 95.0%, and the RSDs were between 4.8% and 8.2%. Hyoscyamine (2 380.0 ng/mL), scopliamine (3.6 ng/mL) and rac-anisodamine (4.7 ng/mL) were detected in the patient's urine. Hyoscyamine (63.3 μg/g), scopliamine (5.7 μg/g) and rac-anisodamine (2.1 μg/g) were detected in eight treasure congee. Hyoscyamine (901.0 μg/g), scopliamine (80.0 μg/g) and rac-anisodamine (30.1 μg/g) were detected in the seed of Datura stramonium L. The ratio of scopliamine and hyoscyamine in the seed of D. stramonium was 1∶11, which complies with the characteristics of D. stramonium L. In urine sample, the proportion of scopliamine and rac-anisodamine was 0.15% and 0.20%, and hyoscyamine accounted for 99.65%. Conclusion Seed morphology, the content range and proportion of three alkaloids are all in accord with the characteristics of D. stramonium. Combined with the clinical symptoms of atropine poisoning, it can be deduced that this incident is a family food poisoning caused by accidental consumption of seed of D. stramonium L. The method can provide technical support for the clinical diagnosis and treatment of alkaloid poisoning patients, and also provide a basis for emergency detection and disposal of alkaloid poisoning events.
9.Effect of modified huanglian wendan decoction in treating senile patients with mild cognitive impairment of turbid-phlegm blocking orifice syndrome.
Ren-zhen GUO ; Wen-quan ZHOU ; Zeng-gang LUO
Chinese Journal of Integrated Traditional and Western Medicine 2010;30(1):33-36
OBJECTIVETo observe the clinical efficacy of modified Huanglian Wendan Decoction (HWD) in treating senile mild cognitive impairment (MCI) of turbid-phlegm blocking orifice syndrome.
METHODSWith a block randomized, double-blinded and controlled design adopted, the 64 patients of MCI selected from December 2007 to February 2009 were randomly and equally assigned to two groups. The treatment group was treated with HWD in dose of 200 mL, twice a day; the control group was given Aniracetam 0.2 g (for patients over 70-years-old, 0.1 g) three times a day. And the illusive medicine in dosage-form of capsule/decoction simulated to that used in the opposite group was applied. The medication and observation lasted for three months. Chinese medicine syndrome, cognition capacity (by MMSE), laboratory indexes [acetylcholine (Ach), superoxide dismutase (SOD), malondialdehyde (MDA)] and safety related indexes in patients were observed.
RESULTSAfter treatment, MMSE score increased in both groups, but the increment in the treatment group was significantly higher than that in the control group (P<0.01); Chinese medicine syndrome estimated by scoring showed that after treatment, all scores of syndromes, excepting the expectoration, were improved in the treatment group with the post-treatment scores significantly lower than those in the control group respectively (P<0.05 or P<0.01); while in the control group, lowering of scores only showed in some symptoms such as poor memory, heavy head or dizziness, and heavy sensation in limbs and body. Serum levels of Ach and SOD decreased and MDA increased in both groups after treatment, but the change of Ach was more significant in the treatment group (P<0.01). No obvious adverse reactions were found during the treatment.
CONCLUSIONFor treatment of MCI, HWD shows effects in improving patients' symptoms, cognition capacity and elevating serum Ach content better than that of Aniracetam; and with effects for raising SOD activity and reducing MDA level similar to those of Aniracetam.
Aged ; Aged, 80 and over ; Cognitive Dysfunction ; diagnosis ; drug therapy ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Pyrrolidinones ; therapeutic use
10.Investigation on the natural infection of Theiler's murine encephalomyelitis virus and study on experimental infection of the virus in mice
Wen YUAN ; Yu ZHANG ; Bihong HUANG ; Yinzhu LUO ; Jing WANG ; Jinchun PAN ; Ruike WU ; Pengju GUO ; Ren HUANG
Chinese Journal of Comparative Medicine 2017;27(4):75-81
Objective To investigate the natural infection of Theiler's murine encephalomyelitis virus (TMEV) in mice,and to survey the distribution of virus in tissues and the changes of serum antibody in the experimentally TMEV-infected mice.Methods Enzyme linked immunosorbent assay (ELISA) and fluorescence quantitative RT-PCR (qRT-PCR) assay were used to detect the antibody and nucleic acid of TMEV in clinical samples.These samples included SPF mice collected from Guangdong area in 2010-2015,mice obtained from a non-barrier laboratory rodent colony,and wild Rattus norvegicus live-trapped around the non-barrier laboratory rodent colony.36 ICR mice were intracerebrally inoculated with TMEV BeAn strain.The clinical signs of the animals were observed daily post-inoculation.Three mice were euthanatized at day 0,3,7,10,17,21,31,39 and 46 post-inoculation (dpi),respectively.Tissue and serum samples were collected for TMEV detection.Results The TMEV antibody and nucleic acid positive rates of SPF mice collected from Guangdong area in 2010-2015 were 5.29% (n=2834) and 27.27% (n=457),respectively.The TMEV antibody and nucleic acid positive rates of the mice obtained from a non-barrier laboratory rodent colony were 71.95% (n=82) and 53.66% (n=82),respectively.The TMEV nucleic acid positive rate of wild Rattus norvegicus was 25.93% (n=27).In the TMEV positive mice,only two mice showed obvious clinical symptoms.The cecal contents,feces and brain samples were the best candidates for qRT-PCR assay.The viral nucleic acid could be detected in the brain,heart,liver,lung and stomach of ICR mice at 3 dpi,but no viral nucleic acid was detected in the spleen,kidney,and cecum.The viruses in liver,heart,lungs and stomach were completely cleared at 10 dpi,and the viruses persisted in the brain throughout the experiment.The TMEV antibody could be detected at 7 dpi,and then the antibody positive rate reached 100% at 17 dpi.The antibody level increased gradually and maintained up to 46 days.ICR mice showed latent infection after TMEV inoculation,with no obvious symptoms and eye pathological changes.Conclusions The experimental mice and wild Rattus norvegicus in Guangdong area are both infected with TMEV,and the infection rate is high.The mice inoculated with TMEV BeAn strain show latent infection.The TMEV antibody produced in mice can be detected at 7 dpi and persisted until the end of the experiment.The viruses are found in the liver,heart,lung and stomach for a short time,but are persisted in the brain for a long time.There is a good consistency of TMEV detection between qRT-PCR and ELISA.The qRT-PCR assay can be used as a powerful complement method for the national standard of laboratory animals.