2.Study on transfection of adeno associated virus 2-ND4 gene into mitochondria
Shuo, YANG ; Lei, LIU ; Han, PEI ; Xing, WAN ; Duoduo, LU ; Weikun, HU ; Bin, LI
Chinese Journal of Experimental Ophthalmology 2014;32(8):693-695
Background Leber hereditary optic neuropathy (LHON) is mitochondrial DNA (mtDNA) disease and mainly leads to optical nerve degeneration.Its primary mechanism is synthesis disorder of DN4 protein due to variation of mtDNA 11778 locus.So to construct a vector with exogenous normal ND4 and transfect into mitochondria is a key of gene therapy for LHON.Objective This study was to investigate the in vitro transfection of adeno-associated virus (AAV)-ND4 gene into mitochondria.Methods Human renal epithelial cell lines transfected adenovirus E1A (293 cells) were regularly cultured and divided into two groups.Framework plasmids of recombinant AAV-ND4 or simple AAV2 were added to the cell medium respectively.The expression of ND4 in cells were located 12,24,36 and 48 hours after transfected by Y03 dual fluorescent quantum dots staining.The positive response for ND4 showed the green fluorescence.Results Cultured 293 cells grew well with 80% confluence.Abundant green fluorescence particles were seen in cytoplasm in the AAV-ND4 transfected group,but only red fluorescence from mitochondrial protein was seen in the simple AAV transfected group under the fluorescence microscope.Conclusions Exogenous ND4 protein can been successfully transfected into mitochondria using the ND4 gene constructed AAV.This result provides experimental evidence for the further study on gene therapy of LHON.
3.Relationship between ERCC2 single nucleotide polymorphisms and male idiopathic infertility in Ningxia.
Chun-Lian LIU ; Hai-Yan JIAO ; Qiang MA ; Xing WU ; Shao-Tong JIA ; Wan-hong JING ; Qing-Hu YANG
National Journal of Andrology 2014;20(5):419-422
OBJECTIVETo explore the influence of the DNA repair gene ERCC2 single nucleotide polymorphisms (SNPs) rs13181, rs1618536, and rs1799793 on male idiopathic infertility in Ningxia, China.
METHODSUsing MassArray, we conducted a case-control study and genotyped three ERCC2 SNPs rs13181, rs1618536, and rs1799793 for 351 males (aged 31.0 +/- 4.2 years) with idiopathic infertility and another 327 normal fertile men (aged 33.0 +/- 5.9 years) as controls.
RESULTSThe ERCC2 AnyG-anyA-anyA genotypes were significantly associated with an increased risk of idiopathic infertility (OR 0.414, 95% CI 0.176 - 0.970), while the three single ERCC2 SNPs rs13181, rs1618536, and rs1799793 showed no significant differences between the cases and controls (P > 0.05).
CONCLUSIONThe ERCC2 SNPs rs13181, rs1618536, and rs1799793 play a role of interaction in male idiopathic infertility in Ningxia, contributing to the risk of the disease.
Adult ; Case-Control Studies ; China ; DNA Repair ; Genotype ; Humans ; Infertility, Male ; genetics ; Male ; Polymorphism, Single Nucleotide ; Xeroderma Pigmentosum Group D Protein ; genetics
4.Study on infiltration glass for machinable-infiltrated-ceramic and on its colorants.
Xiaoyu YANG ; Yunmao LIAO ; Suqin XIAN ; Xing LIANG ; Wei WEI ; Meihua WAN
Journal of Biomedical Engineering 2003;20(4):664-667
In order to develop tinted infiltration glass and its colorants, which can make the ceramic have good spectrum transmittance, color space, color stability and be suitable for clinical use, we selected the best prescription and confirm the ingredient and content of the colorants. Molten glass was prepared in Al2O3 crucible by heating the components to 1420 degrees C for 2 hours. The refractive index and thermal stability of the glass were investigated. The refractive index of the MIC infiltration glass was 1.5969(587.6 nm, nd). It was close to the index of aluminous matrix 1.759(546.07 nm, ne), which increased the spectrum transmittance of MIC. The thermal expansion coefficient of the glass was 7.565 microns/m/degree C, which was compatible with the thermal expansion coefficient of aluminous matrix(8.214 microns/m/degree C). This study proved that the tinted infiltration glass has good color stability, spectrum transmittance, and thermal expansion properties.
Aluminum Silicates
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Coloring Agents
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Crystallization
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Dental Porcelain
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Elasticity
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Glass
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Hardness
5.Professor LIU Yanxia's Experience in Treating Gynecological Diseases with Chinese Medicine Application Therapy
Journal of Zhejiang Chinese Medical University 2024;48(1):46-49,58
[Objective]To summarize Professor LIU Yanxia's academic experience in clinical application of Chinese medicine application therapy in the treatment of various gynecological diseases.[Methods]To review and analyze Professor LIU's clinical experience in the treatment of gynecological refractory diseases by using Chinese medicine patch for lower abdomen and vulva application,and systematically summarize Professor LIU's clinical experience from the aspects of etiology,pathogenesis,treatment principles,methods and empirical prescriptions.A medical case of female vulvar lichen sclerosus was attached for further evidence and explanation.[Results]Professor LIU bases on the circulation characteristics of the lower abdominal meridians,and in accordance with the characteristics of etiology and pathogenesis,forms an empirical formula for applying Chinese medicine patch to the lower abdomen,represented by"warming the kidney and warming the uterus""removing stasis and relieving pain""strengthening the spleen and removing dampness""strengthening the body and preventing infection",which has been respectively used to treat infertility,gynecological pain,obesity and sub health conditions.At the same time,Professor LIU divides vulvar lichen sclerosus lesions into proliferative and atrophic types based on the different characteristics,and uses different empirical formulas for vulvar plaster therapy.The case of atrophic vulvar lichen sclerosus was characterized by liver and kidney deficiency syndrome.The overall treatment principle was to tonify the kidney,nourish blood,and eliminate blood stasis and relieve itching,and modified Erxian Decoction and Danggui Buxue Decoction was used,combined with Chinese medicine application therapy for removing stasis and relieving itching at the local area of the vulvar lesion,which resulted in a significant therapeutic effect.[Conclusion]Professor LIU Yanxia has developed an empirical prescription for applying Chinese medicine patch to different diseases based on the different etiology,pathogenesis and characteristics of the disease.The medication features are distinctive,and treatment effect is satisfactory,which can provide reference for clinical practice.
6.Relativity of commercial specification of Menthae Herba based on chemical analysis.
Dan YE ; Ming ZHAO ; Yang SHAO ; Zhen OUYANG ; Hua-sheng PENG ; Han BANG-XING ; Wei-wan-qi ZHANG ; Xue-mei GU
China Journal of Chinese Materia Medica 2015;40(2):251-257
In order to compare the differences of 35 Menthae Herba samples collected on the market and at producing areas, the contents of six total terpenoids, the essential oil and chromatographic fingerprints were analyzed, which provided evidences for drawing up the commodity specifications and grading criteria of Menthae Herba. GC-MS method was used to analyze the chemical constituents of 35 different samples. The chromatographic fingerprints obtained by using GC were then evaluated by similarity analysis, hierarchical clustering analysis and principal component analysis. The relativity between the content of six terpenoids and the essential oil were studied. In this study, the chemical profiles of 35 samples from different producing areas had significant disparity. All samples collected in the report could be categorized into four chemical types, L-menthol, pulegone, carvone and L-menthone, but the chemical profiles had no relationship with the areas. The chromatographic fingerprints of the samples from different types were dissimilar, while the different producing areas were difficult to be separated. It was indicated that the content of volatile oil was positively correlated with the content of L-menthol and the sum of six total terpenoids. The content of the essential oil, L-menthol and the sum of six total terpenoids of Menthae Herba were considered as one of the commercial specifications and grading criteria. These results in the research could be helpful to draw up the commercial specification and grading criteria of Menthae Herba from a view of chemical information.
Cluster Analysis
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Gas Chromatography-Mass Spectrometry
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Mentha
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chemistry
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Oils, Volatile
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analysis
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Principal Component Analysis
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Terpenes
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analysis
7.Protective effect of Shenxiong injection on cerebral ischemia-reperfusion injury of rats.
Xing-Yu WANG ; Yu-Yan ZHANG ; Hai-Tong WAN ; Tao ZHAO ; Wei FU ; Hui-Fen ZHOU ; Peng ZHOU ; Ze-Feng WANG ; Jie-Hong YANG
China Journal of Chinese Materia Medica 2014;39(3):503-506
OBJECTIVETo study the protective effect of Shenxiong injection on the cerebral ischemia-reperfusion injury of senile rats.
METHODTotally 108 Sprague-Dawley (SD) rats were randomly divided into the sham operation group, the model group, the Ni-modipine group and Shenxiong injection groups (low, middle, and high doses). The rat brain ischemia-reperfusion model was established by the middle cerebral artery occlusion (MCAO) method in rats, in order to observe the effect of Shenxiong injection on neurological score and brain infarct volume of rats with cerebral ischemia-reperfusion injury, and determine the contents of NOS, NO, SOD, MDA and LDH in brain tissues. The contents of TNF-alpha and IL-1beta levels in brain tissues were measured by enzyme-linked immunosorbent assay (ELISA) method.
RESULTShenxiong injection could significantly decrease neurological score, injury degree of brain tissues and brain infarct volume of rats with cerebral ischemia-reperfusion injury, increase the vigor of SOD, decrease the levels of MDA, NO, NOS and LDH, and inhibit IL-1beta and TNF-alpha expressions.
CONCLUSIONShenxiong injection has the obvious protective effect on the brain ischemia-reperfusion injury in rats. Its mechanism may be related to the improvement of neurological function, the reduction of free radical injury, and the inhibition of inflammation factor expression.
Animals ; Brain ; blood supply ; drug effects ; metabolism ; Brain Ischemia ; complications ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Injections ; L-Lactate Dehydrogenase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; complications ; drug therapy ; enzymology ; metabolism ; Superoxide Dismutase ; metabolism
8.Mutation analysis and prenatal diagnosis of families affected with Duchenne and Becker muscular dystrophy.
Wan-jun WANG ; Hai-yan ZHU ; Rui-fang ZHU ; Ying YANG ; Xiang-yu ZHU ; Hong-lei DUAN ; Ying ZHANG ; Xing WU
Chinese Journal of Medical Genetics 2013;30(1):45-48
OBJECTIVETo detect potential mutations for probands from families affected with Duchenne/Becker muscular dystrophy (DMD/BMD), and to carry out prenatal diagnosis through identification of female carriers.
METHODSA total of 43 DMD/BMD families were recruited. Multiplex PCR was used to analyze 18 exons within hotspots for DMD gene deletions. Multiplex ligation-dependent probe amplification (MLPA) was used to detect potential deletions and duplications of DMD gene for 43 patients and 36 females from 32 families. Prenatal diagnosis was performed for 27 families.
RESULTSDeletional mutations were detected in 26 patients with multiplex PCR. In addition, MLPA has detected 3 deletions and 6 duplicational mutations, and the ranges of mutations were all determined. Among 36 female members, 18 were determined as carriers of deletional mutations, 10 were excluded as mutation carriers. The status of remaining 8 could not be determined. For prenatal diagnosis, 3 out of 18 male fetuses were diagnosed as patients and 1 female fetus was identified as carrier.
CONCLUSIONMLPA is an accurate and reliable method for detecting deletional/duplicational mutations of DMD gene as well as for prenatal diagnosis and detection of female carriers.
Adolescent ; Child ; Child, Preschool ; Dystrophin ; genetics ; Female ; Heterozygote ; Humans ; Infant ; Male ; Multiplex Polymerase Chain Reaction ; Muscular Dystrophy, Duchenne ; diagnosis ; genetics ; Mutation ; Pedigree ; Pregnancy ; Prenatal Diagnosis
9.Clinical study of chest contour deformity after harvesting of costal cartilage for total ear reconstruction.
Wan-Hou GUO ; Qing-Hua YANG ; Hai-Yue JIANG ; Hong-Xing ZHUANG
Chinese Journal of Plastic Surgery 2008;24(5):365-367
OBJECTIVETo investigate the chest contour deformities after harvesting of costal cartilage for total ear reconstruction, so as to prevent donor site morbidity.
METHODS107 cases of microtia who underwent ear reconstruction with costal cartilage during the period from 1994 to 2003, were retrospectively studied. Both the chest depression deformity and the costal arch deformity were evaluated. The possible related factors to the chest contour deformities were analyzed statistically with Stata software.
RESULTSThe incidence of chest depression was 54.55% (48/88), including 35 cases of mild depression (39.77%) and 13 cases of severe depression (14.77% ). The incidence of costal arch deformity was 52.77% (46/88), including 33 cases of mild deformity (37.50%) and 13 cases of severe deformity (14.77%). The incidence of both deformities was significantly different at different age (P < 0.01), but not at different follow-up period (P = 0.294 and P = 0.225, respectively). Gender was related to the incidence of chest depression (P < 0.01), but not to the costal arch contour deformity (P = 0.619). Comparing resection of the 6th and 7th rib cartilage with resection of 7th and 8th rib cartilage, the incidence of costal arch deformity was significantly different (P < 0.01), but the incidence of depression deformity was not (P = 0.657).
CONCLUSIONSThe incidence of chest deformities increases with early operation time (< 13 years old). The male patients have higher incidence of depression deformity than female. The arch deformity is higher when the 6th and 7th rib cartilage were resected. The follow-up period is not related to the chest deformities.
Adolescent ; Adult ; Child ; Child, Preschool ; Ear, External ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Postoperative Complications ; Reconstructive Surgical Procedures ; adverse effects ; Retrospective Studies ; Ribs ; abnormalities ; transplantation ; Sex Factors ; Thorax ; abnormalities ; Young Adult
10.The predictive value of plasma visfatin and adiponectin in early atherosclerosis in patients with type 2 diabetes
Weidong WANG ; Qinjuan YANG ; Baohong WAN ; Ling XING
Chinese Journal of Geriatrics 2017;36(12):1304-1307
Objective To evaluate the association between plasma levels of visfatin and adiponectin with carotid arterial intima-media thickness in patients with type 2 diabetes.Methods A total of 100 elderly patients with type 2 diabetes and 30 healthy elderly controls were recruited from August 2015 to August 2016 at our hospital.Based on carotid arterial intima-media thickness(IMT) assessed via color Doppler ultrasonography,patients with diabetes were further divided into a thickened IMT group (40 cases)and a non-thickened IMT group (60 cases).Plasma levels of visfatin and adiponectin were analyzed and compared,and their associations with carotid arterial IMT were evaluated.Results Compared with the control group,patients in both the thickened IMT group and the non-thickened IMT group had significantly higher serum levels of visfatin(19.1 ± 2.8)μg/L vs.(40.6±3.9)μg/L,(28.9±3.6)μg/L,which was markedly higher in the thickened IMT group than in the non-thickened IMT group(P<0.05).In addition,compared with the control group,patients in the thickened IMT group and the non-thickened IMT group had significantly lower serum levels of adiponectin(1 590±330)mg/L,(610±170)mg/L,(930±210)mg/L,which was markedly lower in the thickened IMT group than in the non-thickened IMT group (P < 0.05).Furthermore,Pearson's analysis showed that the plasma level of visfatin was positively correlated with carotid arterial IMT(r =0.721,P<0.05),and the plasma level of adiponectin was negatively correlated with carotid arterial IMT(r=-0.688,P<0.05).Conclusions Plasma levels of visfatin and adiponectin are closely related to carotid arterial IMT in patients with type 2 diabetes,andmay be used as plasma markers for the prediction of early atherosclerosis.