2.The protective effect of bone marrow mesenchymal stem cell on lung injury induced by vibrio vulnificus sepsis
Xiao CHEN ; Huan LIANG ; Jie LIAN ; Yang LU ; Xiaolin LI ; Shaoce ZHI ; Guangju ZHAO ; Guangliang HONG ; Qiaomeng QIU ; Zhongqiu LU
Chinese Critical Care Medicine 2014;(11):821-826
Objective To discuss the protective effect of bone marrow mesenchymal stem cell(BMSC)on lung injury induced by vibrio vulnificus sepsis and its mechanism. Methods BMSCs were isolated by whole bone marrow adherent culture from mouse. Male ICR mice were randomly divided into normal saline control group(NS group),normal saline+BMSC control group(NSB group),vibrio vulnificus sepsis group(VV group),vibrio vulnificus sepsis + BMSC group(VVB group)according to random number table,with 40 mice in each group. Sepsis mouse model was reproduced by injecting vibrio vulnificus(1×107 cfu/mL)5 mL/kg through the left side peritoneal cavity, and caudal intravenous injection of BMSC(4×105 cfu/mL)5 mL/kg for intervention after model reproduction. Ten mice in each group were sacrificed at 6,12,24 or 48 hours after injecting vibiro vulnificus,and their lung tissues were harvested. The lung wet/dry(W/D)ratio was calculated. The expression of nuclear factor-κBp65(NF-κBp65)in nucleus was measured by Western Blot. The levels of tumor necrosis factor-α(TNF-α)and interleukins(IL-1β, IL-6)in lung tissue were detected by enzyme-linked immunosorbent assay(ELISA). The pathological changes in lung tissue were observed after hematoxylin-eosin(HE)staining and uranyl acetate-lead citrate staining. Results After vibrio vulnificus injection,lung W/D ratio,the expression of NF-κBp65 in nucleus,and the levels of TNF-α, IL-1β,IL-6 in the lung tissues were significantly increased in VV group compared with those in NS group at all the time points,and peaked at 12 hours. Compared with the VV group,the VVB group had significantly decreased levels of lung W/D ratio,NF-κBp65 expression,and the levels of TNF-α,IL-1β,IL-6,with significant differences at all the time points〔VV group vs. NS group at 12 hours:lung W/D ratio 7.22±0.03 vs. 5.21±0.02,NF-κBp65 expression (glay scale)1.86±0.74 vs. 0.75±0.07,TNF-α(ng/L)433.24±3.23 vs. 106.57±1.21,IL-1β(ng/L)35.64±0.15 vs. 10.64±0.48,IL-6(ng/L)58.84±0.55 vs. 17.69±1.35,all P<0.05;VVB group vs. VV group at 12 hours:lung W/D ratio 6.49±0.06 vs. 7.22±0.03,NF-κBp65 expression(A value)1.16±0.08 vs. 1.86±0.74,TNF-α(ng/L)357.22±3.25 vs. 433.24±3.23,IL-1β(ng/L)27.77±0.59 vs. 35.64±0.15,IL-6(ng/L)38.68±1.29 vs. 58.84±0.55,all P<0.05〕. There were no significant differences in above indexes between NS group and NSB group. In the NS and NSB groups pathological changes were not obvious under light microscopy,in the VV group lung tissue hyperemia and edema was significant,the edema fluid,red blood cells and inflammatory cells also could be seen, and in the VVB group lung damage that mentioned above could be alleviated. In the NS and NSB groups epithelial cell structure of type Ⅰ and type Ⅱ was completed,and the changes were not obvious under the transmission electron microscopy. In the VV group the alveolar walls were damaged significantly,with type Ⅰ epithelial cell cytoplasm swelling,bubbling and rupture,with type Ⅱ epithelial cells visible cytoplasm decrease,cavitation,addiction to osmium lamellar corpuscle emptying,lysosome hyperplasia,microvilli reduction,and in the VVB group the above damage was alleviated. Conclusion Vibrio vulnificus sepsis can cause acute lung damage and edema,and BMSC can down regulate inflammatory cytokines,reduce lung injury caused by vibrio vulnificus sepsis.
3.Study on genetic approach in the detection of Cryptosporidium parvum and Giardia lamblia in acquired immunodeficiency syndrome patients.
Si-qi LU ; Feng-yun WANG ; Ke ZHANG ; Lian-zhi XU
Chinese Journal of Epidemiology 2006;27(10):884-888
OBJECTIVETo establish genetic method in detecting Cryptosporidium parvum and Giardia lamblia which often coinfected with AIDS patients.
METHODSCryptosporidium oocysts and Giardia cysts were isolated and purified from fecal samples of the individuals infected with C. parvum and G. lamblia, respectively. Genomic DNAs were extracted. Two pairs of specific primers were designed or synthesized according to the 18S rRNA gene from C. parvum or the triose phosphate isomerase (tim ) gene from G. lamblia. Polymerase chain reaction(PCR) technique was used to amplify the DNA samples from the oocysts and the cysts, and those from the 6 control samples, including Schitosoma japonicum, Toxoplasma gondii , Entamoeba histolytica, Trichinella spiralis, Trichomonas vaginalis and human blood cells. DNA samples from 30 fecal samples of AIDS patients were detected with the same method.
RESULTSOne fragment of 500 bp was amplified with the primer of C. parvum, and the other one of 683 bp was amplified with the primer of G. lamblia. Twenty pg and 0.4 pg DNA of C. parvum and G. lamblia could be detected separately. The specificity of these two pairs of PCR primers was confirmed by the failure in the amplification of the control DNA samples. Out of 30 cases of AIDS patients, 7 showed C. parvum positive, while non Giardia was detected.
CONCLUSIONGenetic detection method for C. parvum and G. lamblia detection was established which was more sensitive and specific.
Acquired Immunodeficiency Syndrome ; microbiology ; Cryptosporidiosis ; diagnosis ; Cryptosporidium parvum ; genetics ; DNA, Bacterial ; Giardia lamblia ; genetics ; Giardiasis ; diagnosis ; Humans ; Polymerase Chain Reaction ; methods ; Sensitivity and Specificity
4.Effect of morphine chloride on contractility of small intestinal muscle in vitro or in vivo and its mechanisms.
Zhi-Ying FENG ; Lian-Gen MAO ; Yuan LU
Journal of Zhejiang University. Medical sciences 2008;37(3):271-275
OBJECTIVETo investigate the effect of morphine chloride on small intestinal muscle in vitro or in vivo and its mechanisms.
METHODSContractile amplitude, tension and frequency of the isolated small intestine of rabbits were measured before and after treatment of morphine chloride. The propulsive distance of magenta in intestinal tract was measured when different concentration of morphine chloride was given orally in mice.
RESULTAfter treatment of different concentration of morphine chloride (5 mg/L, 10 mg/L, 30 mg/L), the contractile activities of isolated small intestines of rabbits decreased significantly. The inhibitory effect of morphine chloride was blocked by naloxone, atropine, but potentiated by regitine. The propulsive distance of magenta in intestinal tract of intact mouse decreased after treatment with morphine chloride of various concentration (75, 150, 300 mg/L).
CONCLUSIONMorphine chloride has an inhibitory effect on the contractility of rabbit small intestine in vitro or in vivo. Opioid receptor, choline and adrenal receptor might be involved in this effect.
Animals ; Female ; Gastrointestinal Transit ; drug effects ; In Vitro Techniques ; Intestine, Small ; drug effects ; Male ; Mice ; Morphine ; pharmacology ; Muscle Contraction ; drug effects ; Muscle, Smooth ; drug effects ; Rabbits
5.Analysis of verrucous plaque-type porokeratosis.
Hong FANG ; Qi-fan SHEN ; Lian-zhi LU
Journal of Zhejiang University. Medical sciences 2003;32(3):264-266
Adolescent
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Adult
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Child
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Child, Preschool
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Female
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Humans
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Laser Coagulation
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Male
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Porokeratosis
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pathology
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surgery
6.Effects of different concentration extract from shenghua decoction on contractile activity of the uterine smooth muscle isolated from normal, estrogen-treated and postpartum mice.
Ding ZHAO ; Wen-Hong ZHAN ; Lian-Huai LI ; Feng-Zhi NIE ; Jun-Jie JIAO ; Yang LU
China Journal of Chinese Materia Medica 2006;31(3):243-246
OBJECTIVETo investigate the effects of different concentration extract from Shenghua decoction on contractile activity of the uterine smooth muscle isolated from normal, estrogen-treated and postpartum mice.
METHODMedlab/4 s vital signal recorder was used to measure the effects of extract from Shenghua decoction (3-12 mg x mL(-1)) on contractile amplitude and frequency of the isolated uterus from normal, estrogen-treated and postpartum mice.
RESULTShenghua decoction extract (3-12 mg x mL(-1)) significantly decreased the contractile activity of the mouse isolated uterus in normal non-pregnancy and postpartum, but significantly increased that of the mouse isolated uterus treated with estrogen, and didn't show significant concentration-response relationship.
CONCLUSIONThe effects of Shenghua decoction extract on contractile activity of mouse-isolated uterus treated with estrogen cannot represent the pharmacological effects on that of in normal non-pregnancy and postpartum uterus.
Angelica sinensis ; chemistry ; Animals ; Dose-Response Relationship, Drug ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Estrogens ; pharmacology ; Female ; In Vitro Techniques ; Mice ; Muscle, Smooth ; drug effects ; Plants, Medicinal ; chemistry ; Postpartum Period ; Uterine Contraction ; drug effects ; Uterus ; drug effects
7.Propylthiouracil-induced overt hepatic injury in patients with hyperthyroidism.
Xiao-lan LIAN ; Yao BAI ; Wei-xin DAI ; Zhi-sheng GUO ; Wei LI ; Lin LU
Acta Academiae Medicinae Sinicae 2004;26(2):172-177
OBJECTIVETo study the incidence, clinical features and related factors of propylthiouracil (PTU)-induced hepatic injury in patients with hyperthyroidism.
METHODSA prospective study were carried out in 70 patients of hyperthyroidism with normal liver function. Every patient was treated with PTU 300 mg/d until the thyroid functions recovered to normal, following by decease and maintenance PTU dose in period of six months. Liver function, including serum levels of alanine aminotransferase (ALT), alkaline phosphatase (ALP), aspartate aminotransferase (AST), total bilirubin (TBIL) and direct bilirubin (DBIL), thyroid function (serum thyroxine, triiodothyronine, free thyroxine, and free triiodothyronine and thyrotropin) and blood routine items were measured before therapy and once a month for six months after PTU therapy was begun.
RESULTSSixty-four cases of 70 patients completed the therapy for 6 months. Hepatic injury developed in 33 patients (51.6%). Asymptomatic, transient hepatic injury was shown in 22 patients (34.4%). Slight symptomatic hepatic injury occured in 6 cases (9.4%) and overt hepatic injury in 5 patients (7.8%) after PTU administration. However, all the patients who developed overt hepatic injury did not stop PTU. Hepatic function returned normal one month after stopping PTU. No one finally suffered from viral hepatitis and autoimmune hepatitis in patients of symptomatic and overt hepatic injury.
CONCLUSIONSPTU-induced symptomatic hepatic injury is not rare and usually develops within the first few months of PTU administration. Its clinical course is relatively benign. However, it may be difficult to predict its development, so all patients should be monitored for liver function test during the administration in early stage.
Adolescent ; Adult ; Aged ; Antithyroid Agents ; adverse effects ; therapeutic use ; Chemical and Drug Induced Liver Injury ; Female ; Follow-Up Studies ; Humans ; Hyperthyroidism ; drug therapy ; Liver ; pathology ; physiopathology ; Liver Diseases ; physiopathology ; Liver Function Tests ; Male ; Middle Aged ; Propylthiouracil ; adverse effects ; therapeutic use ; Prospective Studies
8.Influence of drug treatment on glucocorticoid receptor levels in patients with coronary heart disease.
Hong JI ; Wei-Zao GUO ; Zhi-Hong YAN ; Di LI ; Cui-Lian LU
Chinese Medical Journal 2010;123(13):1685-1689
BACKGROUNDGlucocorticoid signaling exerts major roles in inflammation, metabolism and depression, which are three crucial factors accompanying or underlying coronary heart disease. Although accumulating evidence indicates the influence of glucocorticoids on the pathology and treatment of coronary heart disease, there is still a dearth of pharmaceutical mechanisms for this relationship. This study aimed to investigate the influence of drug treatment on glucocorticoid receptor levels in coronary heart disease.
METHODSEighty hospitalized patients (average age (59.0 +/- 7.5) years, 46 male and 34 female) with coronary heart disease were categorized into four groups with 20 members in each according to one of the four drugs they were treated with. The four drugs were: nitrated derivative isosorbide dinitrate, the beta-adrenergic receptor blocker metoprolol, the calcium antagonist nifedipine, and the HMG-CoA reductase inhibitor lovastatin. Glucocorticoid receptor protein levels of peripheral blood lymphocytes were tested using immunoblotting analysis before and after one month of treatment.
RESULTSImmunoblotting analysis showed increased glucocorticoid receptor levels after treatment with metoprolol and nifedipine. There were no statistically significant changes of glucocorticoid receptor levels after treatment with isosorbide dinitrate or lovastatin, although there were trends of up-regulation of glucocorticoid receptor expression after both treatments.
CONCLUSIONSBoth the beta-blocker and the calcium blocker can increase glucocorticoid receptor levels after chronic administration. This effect suggests a mechanism for their anti-inflammatory and other therapeutic roles for coronary heart disease and comorbid disorders.
Aged ; Blotting, Western ; Coronary Disease ; drug therapy ; metabolism ; Female ; Humans ; Isosorbide Dinitrate ; therapeutic use ; Lovastatin ; therapeutic use ; Male ; Metoprolol ; therapeutic use ; Middle Aged ; Nifedipine ; therapeutic use ; Receptors, Glucocorticoid ; metabolism
9.Study on effects of sulfur fumigation on chemical constituents of Chrysanthemum morifolium cv. Boju.
You-lian LI ; Shan WANG ; Jing-jing ZHU ; Wei-hao WANG ; Shi-xie XIANG ; Wei-hong FENG ; Liang-mian CHEN ; Zhi-min WANG ; Tu-lin LU
China Journal of Chinese Materia Medica 2015;40(13):2624-2628
A comprehensively comparison of the chemical profiles between sun-drying BJ (NBJ) and sulfur-fumigated BJ (SBJ) was conducted by HPLC analysis and the discrepant peaks were identified or tentatively assigned by HPLC-ESI-MSn. A total of 32 chemical components were used for qualitative comparison. Meanwhile, a quantitative comparison of BJwere conducted by HPLC analysis and determining seven compounds from 3 NBJ and 3 SBJ samples dramatic chemical changes were found. After sulfur fumigation, the contents of flavonoids glycosides and phenolic acids were remarkably reduced, but the contents of flavonoids aglycones were significantly increased. Multivariate statistics, including principle component analysis (PCA) and partial least squares discriminate analysis (PLS-DA) were used to investigate the potential damaging effect of sulfur-fumigating process. The PCA score plots showed six samples were clearly classified into the sun-drying and sulfur-fumigating groups. And according to VIP >1, the most important chemical markers were apigenin, luteolin and 3,5-dicaffeoylquninic acid which could be used to distinguish NBJ and SBJ samples. Combining the results of qualitative and quantitative analysis, it showed that the sulfur fumigation has a significant effect on BJ.
Chromatography, High Pressure Liquid
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Chrysanthemum
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chemistry
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Fumigation
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Least-Squares Analysis
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Principal Component Analysis
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Sulfur
10.Lovastatin changes activities of lactate dehydrogenase A and B genes in rat myocardial cells.
Wei-Zao GUO ; Hong JI ; Zhi-Hong YAN ; Lin LI ; Di LI ; Cui-Lian LU
Chinese Medical Journal 2011;124(3):423-428
BACKGROUNDLactate dehydrogenase (LDH) is a crucial regulator of energy metabolism in many organs including the heart. Lovastatin is widely used in prevention and treatment of coronary heart disease and is a drug with substantial metabolic influences. Our study aimed to determine the activities of the lactate dehydrogenase A and B (LDHA and LDHB) genes following lovastatin treatment.
METHODSThe rat myocardial cell line H9c2(2-1) in culture was exposed to 100 nmol/L lovastatin for 24 hours or for five days. The functions of the LDHA and LDHB genes were examined at the transcriptional (mRNA) level with quantitative real-time polymerase chain reaction (Q-RT-PCR), and at the translational (protein) level with immunoblotting.
RESULTSWhen compared with control levels, the LDHA mRNA went up by (151.65 ± 16.72)% (P = 0.0132) after 24 hours and by (175.28 ± 56.54)% (P = 0.0366) after five days of lovastatin treatment. Although 24 hours of lovastatin treatment had no significant effects on LDHB mRNA levels, when the treatment was extended to five days, LDHB mRNA levels were significantly down-regulated to (63.65 ± 15.21)% of control levels (P = 0.0117). After 24 hours of treatment with lovastatin, there were no significant changes in protein levels of either LDHA or LDHB. When treatment time was extended to five days, the protein levels of LDHA were up-regulated by (148.65 ± 11.81)% (P = 0.00969), while the protein levels of LDHB were down-regulated to (64.91 ± 5.47)% of control levels (P = 0.0192).
CONCLUSIONSLovastatin affects gene activities of LDHA and LDHB differently, which may reveal novel pharmacological effects of lovastatin.
Animals ; Anticholesteremic Agents ; pharmacology ; Blotting, Western ; Cell Line ; Isoenzymes ; genetics ; metabolism ; L-Lactate Dehydrogenase ; genetics ; metabolism ; Lovastatin ; pharmacology ; Myocytes, Cardiac ; drug effects ; enzymology ; Rats ; Reverse Transcriptase Polymerase Chain Reaction