1.Effects of Ding Kundan on expression of TGF-β1,CTGF and AR in PCOS model rats
Lan CHEN ; Yong TAN ; Shu-Ping CHEN
Chinese Journal of Immunology 2018;34(2):218-222
Objective:To investigate the effect of Ding Kundan on the expression of TGF-β1,CTGF and AR in PCOS model rats,and to explore its treat mechanism on PCOS.Methods: The PCOS model rats induced by DHEA were intervened with high, medium and low dose of Ding Kundan(4.5,2.25,1.125 g/kg),ELISA,immunohistochemistry,RT-PCR and Western blot were used to detect serum hormone levels,mRNA and protein expression of TGF-β1,CTGF and AR in each group of rats.Results: Compared with model group Ding Kundan high,medium and low dose group of E2,LH and T were decreased,while FSH was increased.And the differences were statistically significant.Protein expression and mRNA of TGF-β1,CTGF and AR were decreased,and the differences were statistically significant.Conclusion: Ding Kundan shows significantly therapeutic efficacy on PCOS rats,and its possible mechanism is related to regulating expression levels of TGF-β1,CTGF and AR in ovarian tissue.
2.Analysis of DNA fingerprint of Mycobacterium tuberculosis enterbacterial repetitive intergenic consensus-polymerase chain reaction
De-Cui PEI ; Qing-Hua LUO ; Xiang WANG ; Shu-Lan WANG ; Ya WANG ; Jin-Yong WANG ;
Chinese Journal of Laboratory Medicine 2003;0(07):-
Objective To analyze the epidemiological characteristics of Mycobacterium tuberculosis by enterbaeterial repetitive intergenic consensus-polymerase chain reaction(ERIC-PCR)DNA fingerprint. Methods Mycobacterium tuberculosis positive sputum samples between September 2003 to May 2006 were collected and cultured.Chromosomal DNA were extracted and ERIC-PCR DNA fingerprinting was analyzed by software,such as RAPD PHYLIP and Treeview.Results A total of 42 different fingerprints were detected.Phylogenetic analysis showed that they could be classified into three clusters,the clustering rate was 72.6%.The characteristics of ERIC-PCR fingerprint patterns were related to age,drug resistance,and type of resistance.Conclusions ERIC-PCR DNA fingerprinting technique used in this study is good for epidemiological studies with its strong discrimination,simplicity and rapidness.A high level of recent transmission is found in our city.
3.Inhibitory effect of eicosapentaenoic acid on proliferation of human vascular endothelial cells induced by vascular endothelial growth factor
Yong-qin, BAO ; Jing-xue, MA ; Shu-fen, WANG ; Lan-cun, L(U) ; Ying-hua, DU
Chinese Journal of Experimental Ophthalmology 2011;29(1):38-42
Background Eicosapentaenoic acid(EPA)function as the critical lipid mediators involved in several biological events in human body and play important role in suppressing the genesis of vascular endothelial growth factor (VEGF),migration and proliferation of vascular endothelial cells.Many ocular diseases were proved to be associated with neovascularization.Objecfive The purpose of this study was to investigate the inhibitory effect of EPA on the proliferation of human umbilical vein endothelial cells (HUVEC) indueed by VEGF. Methods HUVEC strain was cultured and passaged,and difierent concentrations of EPA were added to the medium with and without VEGF.The cultured cells were identified by antiofactor Ⅷ polyclonal antibody.The suppressing role of different concentrations of EPA on the proliferation of VEGF-induced or-uninduced HUVEC was assessed by MTT method.The influence of difierent concentrations of EPA on the cellular cycle of VEGF-induced HUVEC was assayed using flow eytometry.The expression of Flk-1,a receptor of VEGF,in the HUVEC Was detected by immunohistochemistry. Results Cultured HUVEC showed the ftlsiform in shape and presented with the cobblestone-like arrangement with the positive response for Ⅷ factor-related antigen.Various concentrations of EPA showed obviously inhibitory effect on VEGF-induced or-unindueed HUVEC at a dose-dependent manner (F=23.072.P=0.000).The inhibitory ability of EPA on VEGF-induced HUVEC was stronger than VEGF-uninduced HUVEC(F=41.417,P=0.000).In 24,48 and 72 hours,the action of EPA on the proliferation of HUVEC was gradually enhanced with the prolong of time(F=1.495,P=0.236).Cell cycle analysis indicated that EPA arrested VEGF-induced HUVEC in G0/G1 phase.The ratio of HUVEC in G0/G1 phase in EPA group was(75.83±1.56)%,and that in control groups was(68.62±1.44)%,showing a significant difference between them(t=-5.88,P=0.00),and no apoptosis of HUVEC was found in both groups.Flk-1 was strongly expressed in the cellular nucleus and cytoplasm in control group.However,the positive expressing intensity of Flk-1 in the HUVEC weakened,and the positive cell number was evidently less in EPA group. Conclusion EPA can inhibit the proliferation of VEGF induced HUVEC through arresting the synthesis of DNA of HUVEC and downregulate the expression of Flk-1 in HUVEC.These results suggest that EPA might exert an antiangiogenic effect.
4.Response of gastric-related neurons in the hypothalamic paraventricular nucleus to acupuncture at Neiguan and Zusanli in a rat model of gastric distension
Shu CHEN ; Chunyan YONG ; Heng CHEN ; Xiao CHU ; Chao ZHANG ; Cheng TAN ; Lan YE ; Jiangshan LI
Chinese Journal of Tissue Engineering Research 2014;(5):675-680
BACKGROUND:Previous studies concerning neuromechanism of acupuncture for regulating gastric function mainly focused on peripheral nerve, involving in central nervous system. In particular, there are few studies addressing higher central nervous system.
OBJECTIVE:To explore the effects of acupuncture at Neiguan (PC6) and Zusanli (ST36) on gastric-related neurons in the hypothalamic paraventricular nucleus.
METHODS:Using microelectrode extracellular recording technique, gastric related neurons in the paraventricular nucleus were found in gastric distension rats. Stimulation was conducted for 30 seconds by hand acupuncture at Neiguan and Zusanli. The effects of acupuncture on gastric-related neurons in the paraventricular nucleus were observed.
RESULTS AND CONCLUSION:The discharge of 109 neurons in the hypothalamic paraventricular nucleus of 60 rats was recorded. A total of 56 gastric-related neurons of the 109 neurons were observed. The number of neurons response to acupunctures at Zusanli, Neiguan, Pishu (BL20) and Weishu (BL21) were respectively 44, 47, 29, and 33, with the reaction proportion of 78.57%, 83.47%, 51.79%and 58.93%. Results suggested that there exist gastric distension stimulation and acupuncture stimulations from neurons soma to visceral reaction in the hypothalamic paraventricular nucleus, which simultaneously participates in the regulation of acupuncture on the gastric function.
5.Gastric distention-related neurons in dorsal nucleus of the vagus nerve of rats in response to different acupuncture acupoints
Shu CHEN ; Chunyan YONG ; Heng CHEN ; Xiao CHU ; Chao ZHANG ; Cheng TAN ; Lan YE ; Jiangshan LI
Chinese Journal of Tissue Engineering Research 2014;(36):5842-5846
BACKGROUND:Preliminary studies of our research group mainly focus on the role of nucleus of solitary tract in gastric noxious stimulation and acupuncture point, while dorsal nucleus of vagus nerve, nucleus of solitary tract and area postrema are the necessary component for vagus nerve complex, and dorsal nucleus of vagus nerve plays a crucial role in the regulation of gastric functions. OBJECTIVE:To observe the effect of acupuncture at Zusanli, Neiguan, Pishu and Shenshu acupoints on the gastric-related neurons in dorsal motor nucleus of the vagus nerve. METHODS:The gastric related neurons in dorsal motor nucleus of the vagus nerve of gastric distension rats were identified using extracellular recording technique, according to neuroelectrophysiology method. Acupuncture stimulation was given to Zusanli, Neiguan, Pishu and Shenshu for 15 seconds. The effect of body stimulation on the gastric distension related neurons in dorsal motor nucleus of the vagus nerve of rats was recorded. RESULTS AND CONCLUSION:Among the involved 24 rats, 90 neurons at paraventricular hypothalamic necleus exhibited discharge. Among the discharged neurons, 54 neurons were related to gastric distension. After acupuncture stimulation at Zusanli, Neiguan, Pishu and Shenshu, there were 44, 39, 38, 27 neurons giving a response, the reaction rate was 82%, 72%, 70%and 50%, respectively. Acupunture at different nerve segment acupoints can activate the dorsal motor nucleus of the vagus nerve to different degrees, which is involved in the regulation of gastric motility.
6.Relationship between the Pattern of Methylation of Calcitonin Gene and Activity of Methyltransferase in 8 Tumor Cell Lines
Xiao-Chuan BAI ; Zhi-Yong BAI ; Shu-Lan WU
Chinese Journal of Cancer 2001;20(2):156-159
Objective: The current study was designed to investigate the relationship between the pattern of methylation of calcitonin (CT) gene and the activity of DNA methyltransferase (MTase). Methods: The methylation rate of 5′ region of CT gene (CTMR) and the activity of MTase in 6 solid tumor cell lines and 2 leukemia cell lines were determined. CTMR was detected by polymerase chain reaction(PCR) with inter and external references in combination with restriction endonuclease and laser scan technique. MTase activity was examined by isotope labeled microassay. Results:Both CTMR and MTase activity in all tumor cell lines were significantly higher than that of control cells. The increased MTase activity was relative to elevated CTMR. Conclusion: There is prevalence of hypermethylation of CT gene and elevated activity of MTase in malignant cells. The increased MTase activity is one of the possible reasons for CT gene hypermethylation in tumor cells.
7.Gain of human telomerase RNA gene is associated with progression of cervical intraepithelial neoplasia grade I or II.
Yong-Lian LAN ; Lan YU ; Chan-Wei JIA ; Yu-Mei WU ; Shu-Yu WANG
Chinese Medical Journal 2012;125(9):1599-1602
BACKGROUNDThe 3q26 chromosome region, where the human telomerase RNA gene (hTERC) is located, is a biomarker for cervical cancer and precancerous lesions. The aim of this study was to confirm the value of measuring hTERC gene gain in predicting the progression of cervical intraepithelial neoplasia grade I or II (CIN-I and -II, respectively) to CIN-III and cervical cancer.
METHODSLiquid-based cytological samples from 54 patients with CIN-I or CIN-II lesions were enrolled in this study. Follow-up was performed with colposcopy and biopsy within 24 months after the diagnosis of CIN-I or CIN-II. Copy numbers of the hTERC gene were measured by fluorescence in situ hybridization with a dual-color probe mix containing the hTERC gene probe (labeled red) and the control, the chromosome 3 centromere-specific probe (labeled green).
RESULTSAll patients whose lesions progressed from CIN-I or CIN-II to CIN-III displayed a gain of the hTERC gene, whereas patients where the hTERC gene was not amplified did not subsequently progress to CIN-III or cervical cancer. The signal ratio pattern per cell was recorded as N:N (green:red). The numbers of cells with the signal ratio pattern of 4:4 or N:≥5 in patients whose lesions progressed to CIN-III were significantly higher than those whose lesions did not progress. Significantly, none of the patients with a 4:4 signal ratio pattern regressed spontaneously.
CONCLUSIONSIn conclusion, measurement of hTERC gene gain in CIN-I or CIN-II patients using liquid-based cytological samples could be a useful biomarker to predict the progression of such cervical lesions. In addition, a 4:4 or N:≥5 signal ratio pattern may indicate the unlikeness of spontaneous regression of CIN-I or CIN-II lesions.
Adult ; Cervical Intraepithelial Neoplasia ; genetics ; pathology ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Middle Aged ; RNA ; genetics ; Telomerase ; genetics
8.Effects of needling acupoints at different nerve segments on oxytocin neurons in rat's hypothalamic paraventricular nucleus and intragastric pressure
Chun-Yan YONG ; Shu CHEN ; Heng CHEN ; Xiao CHU ; Chao ZHANG ; Cheng TAN ; Lan YE ; Jiang-Shan LI
Journal of Acupuncture and Tuina Science 2019;17(5):297-304
Objective:To compare and explore the effects of needling acupoints at different nerve segmentson the oxytocin (OT) neurons in the paraventricular nucleus of hypothalamus (PVN) and the intragastric pressure, and discuss the possible mechanisms. Methods: Thirty-two healthy adult Sprague-Dawley (SD) rats were numbered and divided into 4 groups according to the random number table, a Zusanli (ST 36) group, a Neiguan (PC 6) group, a Weishu (BL 21) group and a control group, with 8 rats in each group. Except the control group, rats in the other three groups received acupuncture at the corresponding acupoints. To observe the differences in double-labeled OT neurons and c-fos neurons of the hypothalamic PVN and the intragastric pressure after acupuncture among the three groups of needling acupoints at different nerve segments. Results:Compared with the control group, the numbers of double-labeled cells in the PVN of the Zusanli (ST 36) group and the Neiguan (PC 6) group decreased significantly, while the intragastric pressure increased significantly (allP<0.05), and the inter-group differences were statistically significant (P<0.05). The intragastric pressure in the Weishu (BL 21) group decreased significantly, and the inter-group difference was statistically significant (P<0.05). Compared with the Weishu (BL 21) group, the numbers of OT/c-fos double-labeled cells in PVN of the Zusanli (ST 36) group and the Neiguan (PC 6) group decreased significantly, and the intragastric pressure increased significantly, the inter-group differences were statistically significant (allP<0.01). Conclusion:Acupoints at different nerve segments have different regulation effects on intragastric pressure. The difference may be related to the different nerve conduction pathways by acupoints at different nerve segments in regulating the intragastric pressure. The PVN may be one common integration center for the regulation of gastric function in the three acupoints [Zusanli (ST 36), Neiguan (PC 6) and Weishu (BL 21)] at different nerve segments.
9.The expression and significance of glucose-6-phosphate isomerase mRNA in peripheral blood mononuclear cells of patients with rheumatoid arthritis
Guan-Fei ZHAO ; Yong-Zhe LI ; Yu-Guo SONG ; Chao-Jun HU ; Da-Wei TONG ; Shu-Lan ZHANG ;
Chinese Journal of Rheumatology 2001;0(04):-
Objective To investigate the mRNA level of glucose-6-phosphate isomerase(GPI)ex- pression in patients with rheumatoid arthritis(RA).Method Reverse transcription-polymerase chain reaction (RT-PCR)was applied to semiquantitatively analyze GPI mRNA expression in the peripheral blood mononu- clear cells(PBMCs)of 30 active RA patients,30 stable RA patients,30 patients with other rheumatic disease and 30 healthy subjects.Results There was statistically significant differences between patients with RA and other rheumatic diseases(P
10.Pathologic feature and diagnosis of pulmonary alveolar proteinosis.
Zhi-lan MENG ; Hong-rui LIU ; Zhi-yong LIANG ; Shu-ying ZHANG
Chinese Journal of Pathology 2005;34(9):575-578
OBJECTIVETo describe the pathologic features and diagnostic algorithm of pulmonary alveolar proteinosis (PAP).
METHODSThirty-nine biopsy and postmortem cases of PAP were studied by light microscopy and histochemical staining using periodic acid-Schiff (with digestion) (PAS-D), mucicarmine (with digestion) (mucicarmine-D) and alcian blue.
RESULTSHistologically, the affected lung tissue displayed the following characteristic features: (1) alveoli and some of the small bronchioles were filled with eosinophilic and fine granular proteinaceous material with needle-like clefts; (2) proteinaceous material was seen admixed with various numbers of degenerated and sometimes exfoliated pneumocytes; (3) pneumocytes were hyperplastic; (4) alveolar capillaries and alveolar septa had become hyperemic, but pulmonary interstitial inflammation was not obvious; (5) no significant inflammation was identified in the bronchial wall; (6) compensating emphysema was noted in the surrounding lung parenchyma. Fragments of eosinophilic, finely granular proteinaceous material with needle-like clefts were also found in the bronchoalveolar lavage fluid under light microscopy. The proteinaceous material was stained red by PAS-D. The staining for mucicarmine-D was negative, while alcian blue staining was either weakly positive (faint blue staining) or negative. Pathologic examination of lung biopsies and bronchoalveolar lavage fluid thus remaines the gold standard for diagnosis of PAP.
CONCLUSIONSIdentification of homogeneous, eosinophilic, finely granular and PAS-D-positive proteinaceous material with needle-like clefts in alveolar spaces or bronchoalveolar lavage fluid is of diagnostic importance in PAP. Bronchoalveolar lavage, being a relatively safe and non-invasive procedure, can be a useful adjunct in arriving at the final conclusion.
Adult ; Bronchoalveolar Lavage ; Bronchoalveolar Lavage Fluid ; cytology ; Female ; Humans ; Lung ; pathology ; Male ; Middle Aged ; Periodic Acid-Schiff Reaction ; Pulmonary Alveolar Proteinosis ; pathology ; therapy ; Pulmonary Alveoli ; pathology