2.Fluorescent measurements of H~+/K~+ exchange in primary cultured inner medullary collecting duct cell monolayers or rabbit kidney
Qianming XIA ; Jian HUANG ; Zhenliang XIAO ; Guisheng QIAN
Chinese Pharmacological Bulletin 1987;0(02):-
AIM To establish the fluorescent measurement technique for H+ /K+ exchange in primary cultured IMCD cell monolayers of rabbit kidney. METHODS To measure H+ /K+ exchange in the primary cultured IMCD cell monolayers under different CO2 concentrations by the BCECF/AM fluorescent probe. RESULTS [pH], recovery rate after acid loaded in the respiratory acidosis group was 0.0620 ? 0.012, and was significantly higher than that of the control group (0.0504 ? 0.009, P 0.05). CONCLUSION The fluorescent measurements of H+ /K+ exchange in primary cultured IMCD cell monolayers of rabbit kidney has many benefits which are convenient manipulation, stable and credit result, economy and safety, this method should further be popularized.
3.Fluorescent measurements of H+/K+ exchange in primary cultured inner medullary collecting duct cell monolayers of rabbit kidney
Qianming XIA ; Jian HUANG ; Zhenliang XIAO ; Guisheng QIAN
Chinese Pharmacological Bulletin 2001;17(2):230-232
AIM To establish the fluorescent measurement technique for H+/K+ exchange in primary cultured IMCD cell monolayers of rabbit kidney. METHODS To measure H+/K+ exchange in the primary cultured IMCD cell monolayers under different CO2 concentrations by the BCECF/AM fluorescent probe.RESULTS [pH]i recovery rate after acid loaded in the respiratory acidosis group was 0.0620±0.012,and was significantly higher than that of the control group(0.0504±0.009,P<0.05);which in the respiratory alkalosis group was 0.0476±0.007, there was no signifacant difference between ALG and CG (P>0.05). CONCLUSION The fluorescent measurements of H+/K+ exchange in primary cultured IMCD cell monolayers of rabbit kidney has many benefits which are convenient manipulation, stable and credit result, economy and safety , this method should further be popularized.
4.Establishment of model of diabetes and lens posterior capsule opacification induced by alloxan in rabbit
Qi, WEI ; Jin-mao, CHEN ; Min-li, HUANG ; Xia, LI ; Jian-feng, HE ; Shao-jian, TAN
Chinese Journal of Experimental Ophthalmology 2011;29(2):130-134
Background The establishment of diabetic animal model is a crucial step for the study about diabetic eye diseases. At present,the main modeling method include the injection of streptozotocin and alloxan. But the shortcoming of the former is an expensive price, and that of the later is high death rate of animals. Objective This experiment was to discuss the way which decrease the death of alloxan-injected animal and explore the effects of high blood glucose on the posterior capsular opacification (PCO). Methods Forty clean healthy male New Zealand white rabbits were randomly divided into 2 groups. 90mg/kg of alloxan were injected via ear vein once in 20 rabbits to create the diabetic animal models,and the equivalent amount of normal saline solution was injected at the same way as normal blood glucose group. The successful models were selected in the animals with the blood glucose level over 12. 0 mmol/L two weeks later, and PCO of lens were graded based on the method of Odrieh under the slit lamp. Extracapsular lens extraction was then performed on the right eye of rabbits in both groups, and the posterior capsules were obtained from these eyes at the 6th, 10th and 14th days after operation. The expression of proliferating cell nuclear antigen ( PCNA ) in posterior capsular lens epithelial cell was detected by immunohistochemistry. Results The modeling successful rate was 70% after injection of alloxan. The body weight of rabbits in high blood glucose group was significantly lowed and the blood glucose was significantly elevated in comparison with normal blood glucose group ( all P<0. 05). Two weeks after surgery ,2 eyes occurred 2 grade of PCO and only one eye showed the 1 grade of PCO in the high blood glucose group. However, 1 grade of PCO was found in 3 eyes in the normal blood glucose group. Biopsy revealed that PCNA was positively expressed in the cell nuclei of LECs in high blood glucose group rather than the normal blood glucose group from the 10th day after surgery. The proliferation index of PCNA was 0. 86±0. 04 and 0. 25±0. 03 respectively in high blood glucose group and normal blood glucose group, showing a significant difference between them (t = -16. 171 ,P = 0. 000). Conclusion Stable diabetic models of rabbits can be created by intravenous injection of 90 mg/kg alloxan. High blood glucose level is one of the important factors for the development of PCO.
5.Type of sperm DNA strand breaks in infertile men and its clinical implication.
Ren-xiong WEI ; Jian-wei CHEN ; Ji-hong HUANG ; Xiao-xia ZHANG ; Yun CUI
National Journal of Andrology 2015;21(7):604-609
OBJECTIVETo observe the characteristics of sperm single-stranded DNA breaks (SSB) and double-stranded DNA breaks (DSB) in infertile men, explore the association of DSB with male infertility, and provide a new observation index and idea for the diagnosis and treatment of the disease.
METHODSThis study involved 60 infertile men (infertility group) and 30 normal healthy males with infertile wives (control group). We comparatively analyzed the seminal parameters of the two groups, determined sperm concentration and viability using the computer aided sperm analysis system, measured the sperm survival rate by hypoosmotic swelling (HOS) test, examined sperm morphology by Diff-Quick staining, and detected sperm DNA damage by two-tail comet assay.
RESULTSNine two-tail comet models were established for detecting sperm DNA integrity. Comparisons between the fertility and control groups showed that the sperm DNA fragmentation index (DFI) was (33.8 ± 13.1) vs (16.3 ± 7.9)% (P < 0.01), the SSB-DFI was (19.2 ± 11.4) vs (14.9 ± 7.6)% (P > 0.05), the SSB-DFI/DFI was (56.8 ± 32.4) vs (91.4 ± 27.8)% (P < 0.01), the DSB-DFI was (23.9 +13.4) vs (6.1 ± 2.7)% (P < 0.01), and the DSB-DFI/DFI was (70.8 ± 19.5) vs (37.4 ± 11.3)% (P < 0.01). The optimal cut-off value of DSB-DFI/DFI in the diagnosis of male infertility was 39.5%, with the AUG, sensitivity, and specificity of 0.969, 98.3%, and 90%; that of DSB-DFI was 15.85%, with the AUC, sensitivity, and specificity of 0.912, 86.7%, and 80%; and that of DFI was 18.65%; with the AUC, sensitivity, and specificity of 0.861, 90%, 70%, respectively. In the infertile men, neither SSB-DFI nor SSB-DFI/DFI exhibited any correlation with semen parameters (P > 0.05); DFI was correlated negatively with the percentage of progressively motile sperm, sperm survival rate, and the percentage of morphologically normal sperm (P < 0.05 or P < 0.01), but not correlated with sperm concentration (P > 0.05); both DSB-DFI and DSB-DFI/DFI showed a negative correlation with sperm concentration, sperm survival rate, and the percentages of progressively motile sperm and morphologically normal sperm (P < 0.05 or P < 0.01).
CONCLUSIONDouble-stranded, rather than single-stranded DNA breaks, may be a factor inducing male infertility. The type of sperm DNA strand damage is of much reference value for the assessment of male fertility.
Case-Control Studies ; Comet Assay ; DNA Breaks, Double-Stranded ; DNA Breaks, Single-Stranded ; DNA Fragmentation ; Fertility ; Humans ; Infertility, Male ; diagnosis ; genetics ; Male ; Semen Analysis ; Sensitivity and Specificity ; Sperm Count ; Spermatozoa ; Staining and Labeling
6.Literature study on species of honeysuckle flower.
Wei ZHANG ; Lu-Qi HUANG ; Chao-Xia LI ; Jian LI ; Rui-Xian ZHANG
China Journal of Chinese Materia Medica 2014;39(12):2239-2245
Honeysuckle flower is a traditional herbal medicine in China Through systemically sorting and studying literature of Chinese medicine, this article pointed out that leech used by the traditional Chinese medicine in ancient time has the features of twist vine, slight purple stem with clothing hair; opposite growing leaves, ovule shape with clothing hair on both side; two flowers growing from one pedicel, labiate corolla with 3.2 cm longth, flower grows from white color to yellow color, each branch axil grows only one pedicel, the involucre is ovoid shape, and the flower season is from mid-March to mid-May. Among all species of caprifoliaceae, only Lonicera japonica Thunb. meets these botanic features. Therefore, L. japonica Thunb. should be used as the orthodox species of herbal honeysuckle flower.
China
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Flowers
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anatomy & histology
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classification
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History, Ancient
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Lonicera
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anatomy & histology
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classification
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Medicine in Literature
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Medicine, Chinese Traditional
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history
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Plants, Medicinal
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anatomy & histology
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classification
7.Expression of interleukin-13 receptor and its relationship to proliferation activity of human gliomas
Zhi-Bai XIA ; Xin-Jian WU ; Tie-Wei QI ; Hai-Jun WANG ; Zheng-Song HUANG ;
Chinese Journal of Microsurgery 2006;0(06):-
Objective To explore the gene expression of interleukin-13 receptor (IL-13R)?2 and its relationship to proliferation activity of human gliomas.Methods The gene expression of IL-13R?in 50 hu- man gliomas,2 malignant human glioma cell lines and 6 normal brain tissues were studied by RT-PCR.Ki-67 labeling index (Ki267 LI) of all sample were detecteded by immunohistochemical staining.Results Only one normal brain tissues expressed very low IL-13R?2 mRNA,whereas 35 (70%) of 50 human brain tumors expressed 1L-13R?2 mRNA.The positive rate and expression level of IL-13R?2 mRNA were increased with the ascending of WHO tumor grade.(former:rs=0.87;letter:rs=0.69,P<0.01).The difference of posi- tive rate and expression level of IL-13R 2?mRNA between the low grade and high grade tumors was statistical- ly significant,the proliferation activity of gliomas evaluated by Ki-67LI (Ki-67 Labeling Index,Ki-67LI) was positively correlated with IL-13R?2 gene expression and the tumor grade.Conclusion In human cerebral gliomas,IL-13R?2 genes may play an role in the malignant progression.The expression level of malignancy in molecular level and selecting the target of gene therapy.
8.Expression of Thraustochytrium sp.FJN-10 ?~4-Desaturase Gene in Saccharomyces cerevisiae
Jin-Qing CHEN ; Xian-Zhang JIANG ; Li-Xia LIU ; Jian-Zhong HUANG ;
Microbiology 1992;0(06):-
1.6 kb ?4-desaturase gene(FAD4)was amplified by PCR using plasmid pGEM-TFAD4 as template.The fragment was subcloned into the HindⅢ/XbaⅠrestriction site of pYES2.0 vector.Recombinant plasmid pYFAD4 was transformed into Saccharomyces cerevisiae strain INVScl for expression.It was found to exhibit ?4-fatty acid desaturase activity in the recombinant S.cerevisiae YFAD4 in the presence of exogenous fatty acid substrate docosapentaenoic acid(100?mol/L)under introduction of GAL1.Expression of the FAD4 under appropriate media and temperature conditions led to the production of DHA and it reached 41.13% of the total yeast fatty acid by GC detection.It was suggested that the protein encoded by FAD4 could specifically catalyze DPA into DHA.
9.Contrast study of assessment of Her2 gene by fluorescence in situ hybridization and immunohistochemistry technique in neoadjuvant chemotherapy breast cancer patients
Juan HUANG ; Xueli HU ; Jian HAI ; Feiyu CHEN ; Ting XIA ; Yaning HE ; Lili TANG
Journal of Chinese Physician 2012;14(1):5-8
ObjectiveTo evaluate the consistency of HER2 gene status before and after neoadjuvant chemotherapy in breast cancer by FISH (fluorescence in situ hybridization) and IHC (immunohistochemistry) techniques,and analyze the factor of the difference in the result and the feasibility of HER2 gene tested by FISH in the neoadjuvant chemotherapy breast cancer patients.Methods FISH and IHC for HER2 gene expression status was performed on the archival paraffin-embedded sections of breast cancer tissues before and after neoadjuvant chemotherapy from 135 Chinese female patients,x2 test of paired comparison of enumeration data and Kappa analysis were used to compare the difference and consistency of this two techniques.ResultsThe detection rate of HER2 status in punctured cancer tissues before neoadjuvant chemotherapy by FISH and HER2 did not show statistical difference in our research while the opposite result were showed in cancer tissues after neoadjuvant chemotherapy.Moreover,the two techniques of HER2 test were less concordant in patients accepted taxanes neoadjuvant chemotherapy than CAF treatment.ConclusionsThe consistency of FISH and IHC techniques of cancer tissues before neoadjuvant chemotherapy gained advantage compared to the ones after neoadjuvant chemotherapy.Patients received neoadjuvant chemotherapy especially taxanes should take the test of HER2 gene status by FISH technique.
10.Lung protective strategies of ventilation in hyaline membrane disease of neonates
Xiufang YANG ; Kaijun ZHENG ; Ronghua XIA ; Jingguo CHEN ; Jian CHEN ; Huijuan HUANG
Chinese Journal of Primary Medicine and Pharmacy 2008;15(12):2002-2004
Objective To explore the protective ventilation strategies in hyaline membmne disease(HMD) of neonates,in order to decrease the incidence of ventilator-associated lung injury(VAU).Methods 68 newborn infants,diagnosed as of neonatal hyaline membrane disease and needed ventilator support,were recruited from the neonatal intensive care unit from 2003 to 2005.Infant with HMD were divided into the conventional ventilation practice group (the control group) and protective ventilation strategies group (the experimental group).The ventilator settinns,artery blood gas analysis,ventilator-associated lung injury and other clinical data were analyzed and compared.Results The incidence of ventilator-associated lung injury in the experimental group wag significantly lower than that in the cntrol group(2.94% VS 17.6%,P<0.05).PaCO2 in experimental group was significantly higher than that in control group.The mortality in the experimental group was significantly lower than that in control group.Condusion The protective ventilation strategies can decrease the incidence of ventilator-associated lung injury in neonatal HMD.the protective ventilation strategies can decrease the mortality in neonatal HMD.