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.Comparison of vascular complications between adults latent autoimmune diabetes and type 2 diabetes
Xiaohua WANG ; Zhiguang ZHOU ; Xia LI ; Hongguang HUANG ; Changluo LI ; Jian LIN ; Lin YANG
Clinical Medicine of China 2008;24(5):422-424
Objective To compare the prevalence of microvascular and macrovascular complications between adult latent autoimmune diabetes(LADA)and type 2 diabetic subjects.Methods A cross-sectional study was performed in 203 LADA(GADA positive)and 203 T2DM(GADA negative)subjects matched with age,gender and duration as well as family history of diabetes.Parameters of microvascular(albuminuria、eyeground microscopy or fundus fluorescence photography,electromyogram)and macrovascular complications(electrocardiogram,blood pressure,blood lipid,body mass index)as well as blood sugar were compared.Results ①Microvascular complications:Compared with T2DM,LADA cases showed higher prevalence of diabetic nephropathy(39.9%vs.28.6%,P<0.05)and similar prevalence in diabetic neuropathy and retinopathy(P>0.05).②Macrovascular complications:LADA cases presented lower prevalence of hypertension(38.9%vs.55.7%,P<0.01),lower prevalence of metabolic syndrome (33.0%vs.45.3%,P<0.01=,but similar orevalence of coronarv heart disease (CHD) and erebral infarction(CVD)(P>0.05).Conclusion LADA patients present less metabolic syndrome,hypertension and more diabetic nephropathy compared with T2DM.
8.Medical students' cognition on laboratory biosafety
Yunshu LI ; Jian XU ; Xia JIANG ; Shijiao ZHAO ; Qun HUANG ; Yachun GUO
Chinese Journal of Infection Control 2017;16(1):73-77
Objective To investigate cognitive status of medical students of a medical university on laboratory biosafety,and provide basic data for laboratory biosafety management in Chinese universities.Methods 900 full-time undergraduate medical students were chosen by cluster random sampling,questionnaires were filled out in by them.Results 900 questionnaires were distributed,877 (98.21%) valid questionnaires were obtained,49.03% (n =430) were from sophomores,50.97 % (n =447) from juniors,148 (16.88 %) students have ever participated in students'scientific research.The overall awareness rate of laboratory biosafety was 58.72%,only 32.16% of students understood the detailed contents of laboratory biosafety regulations,only 8.21% of students have received training in laboratory biosafety;the awareness rate of laboratory biosafety cabinet was only 14.14%,only 7.75% of students knew which operation should be performed in biosafety cabinet;28.28% of students could deal with waste according to the rules,68.19% of students were able to identify warning signs of biological hazard;92.82% of the students thought that laboratory biosafety-related courses should be set up.The overall awareness rate of laboratory biosafety knowledge and safety behavior was low,which were 42.65% and 41.96% respectively,juniors was higher than that of sophomores(P<0.05);in the aspect of chemical hazards and biological hazards,students with scientific research experience scored higher than those who did not participate in scientific research(all P<0.05).Conclusion Medical students' cognition on knowledge of laboratory biosafety is not optimistic,it is imperative to strengthen the management of education and publicity of laboratory biosafety.
9.Efficient isolation of bovine keratocytes utilizing two step enzymatic digestion
Jie, LI ; Xia, LI ; Shao-jian, TAN ; Bao-yu, HUANG ; Wei-wei, ZHOU ; Ying-ying, CHEN
Chinese Journal of Experimental Ophthalmology 2011;29(5):398-401
Background Efficient and lowcost way to isolate keratocytes is helpful for research on cornea.Either relatively expensive or inefficient is the shortage of those means now applied,while raising the keratocytes through passage will change the phenotype of them quickly.Our aim is to approach the way getting keratocytes effectively utilizing modified two step enzymatic digestion by type I collagenase. Objective To evaluate the effect of isolating the bovine keratocytes utilizing two step enzymatic digestion and observe the morphological changes of the keratocytes during cultivation in vitro. Methods Keratocytes were isolated from bovine corneas using 0.5 mg/mL and 1 mg/mL type I collagenase digestion.The harvesting rate and viability rate of the primary keratocytes were evaluated.During the primary cultivation in vitro,the morphological changes of the keratocytes and their F-action distribution were observed.Results(1)The extracellular matrix of the bovine corneas were almost dissolved by the two step enzymatic digestion,followed the keratocytes completely isolated from the solid matrix.The amount of the harvested keratocytes from each cornea was(2.11±0.15)X106 on average while the viability rate was(91.69±3.73)% and the inoculation rate Was(81.20±1.25)%.(2)The primary keratocytes attached and spreaded out with dendritic and stellate morphology.After 3 days cultured,the branches of the keratocytes were contacting and formed networks.The F-actin detected by phalloidin binding showed a limited cortical localization. Conclusion (1)The method of two step enzymatic digestion can make the extracellular matrix of bovine cornea stroma completely degraded with the advantages in high efficiency of harvesting keratocytes and high cell viability and relatively simple manipulation. (2) The primary bovine keratocytes have dendritic morphology and with limited F-action distribute in cellular cortex.
10.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.