2.Effects of bone marrow stromal cells on endothelial cells proliferation and microvessel formation in vitro
Peng-fei ZHANG ; Ya-zhuo ZHANG ; Qing-guo LI ; Meizhen SUN ; Hongyun WANG ; Le HE
Chinese Journal of Rehabilitation Theory and Practice 2006;12(1):14-15
ObjectiveTo observe the effects of bone marrow stromal cells (BMSCs) on vessel endothelial cells proliferation and microvessel formation in vitro.MethodsBMSCs and brain vessel endothelial cells were separated from adult and divided into co-culture group of BMSCs and endothelial cells, medium group of BMSCs, comparison group. Endothelial cells proliferation and microvessel formation were observed. ResultsEndothelial cells were promoted to proliferate and formate the microvessel in medium group and co-culture group. And the effect was prominence in co-culture group.ConclusionBMSCs can promote the proliferation and microvessel formation of endothelial cells.
3.Clinical Analysis of Human Papillomavirus in Paraffin-embedded Cervical Lesion Tissues by Gene Clip Technology
Wen-Sheng FAN ; Ya-Li LI ; Yi-Zhuo YANG ; Yali LV ; Mei ZHONG ; Ai-Jun LIU ; Lin LIU ;
Chinese Journal of Nosocomiology 2006;0(07):-
OBJECTIVE To investigate the clinical significance of human papillomavirus in paraffin-embedded cervical cancer and precancerous lesion tissue by gene clip technology.METHODS 153 Patients with paraffin-embedded examples.DNA was extracted from paraffin-embedded tissues and amplified by polymerase chain reaction(PCR).RESULTS The positive rate of high-risk HPV of inflammation was 8.33%,CINⅠ45.83%,CINⅡ/CINⅢ 87.50% and invasive cancer 92.21%.The HPV infection rate of squamous cell carcinoma was 94.12%.The HPV infection rate of adenocarcinoma was 88.46%.Among all the patients with cervical cancer and CIN,the infection rate of HPV16,the most genotype,was 88.98%.The infection rate of HPV18,the second most subtype,was 33.06%.In addition,the minority were infected HPV52、33、59、68.Among 48 cases of cervical squamous cell carcinoma,the infection rate of HPV16,HPV18 was 93.73% and 27.08% respectively.Among 23 cases of cervical adenocarcinoma,the infection rate of HPV16,HPV18 was 82.61% and 52.17% respectively.On the other hand,all the patients with cervicitis were HPV single infection.The HPV multiple infection rate of CINⅠ,CINⅡ/CINⅢ,cervical cancer was 20.00%,28.57%,36.62% respectively.CONCLUSIONS Gene chip technology can detect multiple HPV genotypes in paraffin-embedded tissues with high sensitivity and specificity,which is useful in the pathogenesis and prevention of cervical cancer.
4.Expression pattern of genes involved in tropane alkaloids biosynthesis and tropane alkaloids accumulation in Atropa belladonna.
Wei QIANG ; Ya-Xiong WANG ; Qiao-Zhuo ZHANG ; Jin-Di LI ; Ke XIA ; Neng-Biao WU ; Zhi-Hua LIAO
China Journal of Chinese Materia Medica 2014;39(1):52-58
Atropa belladonna is a medicinal plant and main commercial source of tropane alkaloids (TAs) including scopolamine and hyoscyamine, which are anticholine drugs widely used clinically. Based on the high throughput transcriptome sequencing results, the digital expression patterns of UniGenes representing 9 structural genes (ODC, ADC, AIH, CPA, SPDS, PMT, CYP80F1, H6H, TRII) involved in TAs biosynthesis were constructed, and simultaneously expression analysis of 4 released genes in NCBI (PMT, CYP80F1, H6H, TRII) for verification was performed using qPCR, as well as the TAs contents detection in 8 different tissues. Digital expression patterns results suggested that the 4 genes including ODC, ADC, AIH and CPA involved in the upstream pathway of TAs, and the 2 branch pathway genes including SPDS and TRII were found to be expressed in all the detected tissues with high expression level in secondary root. While the 3 TAs-pathway-specific genes including PMT, CYP80F1, H6H were only expressed in secondary roots and primary roots, mainly in secondary roots. The qPCR detection results of PMT, CYP80F1 and H6H were consistent with the digital expression patterns, but their expression levels in primary root were too low to be detected. The highest content of hyoscyamine was found in tender stems (3.364 mg x g(-1)), followed by tender leaves (1.526 mg x g(-1)), roots (1.598 mg x g(-1)), young fruits (1.271 mg x g(-1)) and fruit sepals (1.413 mg x g(-1)). The highest content of scopolamine was detected in fruit sepals (1.003 mg x g(-1)), then followed by tender stems (0.600 mg x g(-1)) and tender leaves (0.601 mg x g(-1)). Both old stems and old leaves had the lowest content of hyoscyamine and scopolamine. The gene expression profile and TAs accumulation indicated that TAs in Atropa belladonna were mainly biosynthesized in secondary root, and then transported and deposited in tender aerial parts. Screening Atropa belladonna secondary root transcriptome database will facilitate unveiling the unknown enzymatic reactions and the mechanisms of transcriptional control.
Alkaloids
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biosynthesis
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genetics
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metabolism
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Atropa belladonna
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genetics
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metabolism
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Gene Expression Regulation, Plant
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genetics
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Hyoscyamine
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genetics
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metabolism
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Plants, Medicinal
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genetics
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metabolism
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Scopolamine Hydrobromide
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metabolism
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Tropanes
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metabolism
5.Effect of moxibustion on expressions of HSP70 mRNA and protein in gastric cancer-bearing rats
Jing TAN ; Ya-Ping LIN ; Shou-Xiang YI ; Huan ZHAO ; Zhuo-Jun PENG ; Li-Zhi OUYANG ; Yan PENG
Journal of Acupuncture and Tuina Science 2019;17(6):395-401
Objective:To observe the effect of moxibustion on the mRNA and protein expressions of heat-shock protein 70 (HSP70) in gastric cancer-bearing rats. Methods: A total of 40 healthy Sprague-Dawley (SD) rats were adaptively fed for one week. The gastric cancer model was prepared by Walker-256 cancer tissue transplantation. After 7 d, 10 rats were randomly selected to verify the successful modeling, and the remaining 30 rats were divided into a model group, a moxibustion group and an infrared group by the random number table method, with 10 rats in each group. After enrollment, the moxibustion group received suspended moxibustion at Zhongwan (CV 12), Guanyuan (CV 4) and bilateral Zusanli (ST 36), (the first group of acupoints) on the 1st day, and suspended moxibustion at bilateral Pishu (BL 20) and Weishu (BL 21), (the second group of acupoints) on the 2nd day, 20 min each time, once a day. Moxibustion was alternately performed every other day at the two groups of acupoints for 21 d. From the day of enrollment, rats in the infrared group were irradiated with the infrared radiation at the stomach area on the 1st day, and at the T12-T13 interspinous region on the 2nd day, 20 min each time, once a day, and the two locations were alternately irradiated every other day for 21 d. During the treatment, rats in the model group were intervened by grasping and fixation without treatment. At the end of the treatment, blood was collected from the inner eye orbit, and the HSP70 expression in peripheral blood was determined by enzyme linked immunosorbent assay (ELISA). Rats were sacrificed, the tumor volume and growth inhibition rate were measured. The position and changes of HSP70 in gastric cancer were observed by streptavidin-perosidase (SP); HSP70 protein expression was determined by ELISA; HSP70 mRNA expression in cancer tissues was determined by reverse transcription-polymerase chain reaction (RT-PCR) assay. Results: In comparison of the model group, the volume growth of the gastric cancer in the moxibustion group was significantly restricted (P<0.01); the volume growth inhibition rate in the moxibustion group was 37.93%; the HSP70 expression in peripheral blood and the cancer tissues was significantly increased (both P<0.01); the expression of HSP70 mRNA and HSP70 content in gastric tumor were both obviously increased in the moxibustion group (P<0.01); and a large amount of HSP70 was released to the outside of cancer cells in the moxibustion group. In comparison of the model group, the volume growth of the gastric cancer in the infrared group was slightly restricted (P<0.05) with a volume growth inhibition rate of 15.89%; the HSP70 expression in the infrared group was increased significantly in peripheral blood (P<0.01) and in the gastric cancer tissues (P<0.05); more HSP70 was released outside of the cancer cells in the infrared group. In comparison of the infrared group, the volume growth of gastric cancer was more restricted in the moxibustion group (P<0.05), and the HSP70 expression in the gastric cancer tissues was also higher (P<0.05); more HSP70 was released outside of the cancer cells in the moxibustion group. Conclusion: Moxibustion and infrared treatment inhibit the gastric cancer growth in the gastric cancer-bearing rats, up-regulate the HSP70 expression in gastric cancer tissues, and promote the production and extracellular release of HSP70, and the effect of moxibustion is more obvious.
6.Effect of moxibustion on T lymphocyte subsets in peripheral blood of rats with gastric cancer
Jing TAN ; Ren-Da YANG ; Huan ZHAO ; Zhuo-Jun PENG ; Li-Zhi OUYANG ; Ya-Ping LIN ; Yan-Ping YANG
Journal of Acupuncture and Tuina Science 2019;17(3):161-167
Objective:To observe the effect of moxibustion on T lymphocyte subsets in peripheral blood of rats with gastric cancer. Methods:Sixty healthy Sprague-Dawley (SD) rats were adaptively fed for one week. By the random number table method, 10 rats were randomly selected as a blank group, and 12 rats were randomly selected to simulate the tumor transplantation process; after transplantation, 10 rats were randomly selected as a sham operation group. The remaining 38 rats were used to prepare gastric cancer models by gastric transplantation of the Walker-256 tumor tissue; 8 rats were randomly selected to verify the successful modeling after 7 d; the remaining 30 rats were randomly divided into a model group, a moxibustion group and an infrared group by the random number table method, with 10 rats in each group. From the first day of enrollment, the rats in the moxibustion group received mild moxibustion at Zhongwan (CV 12), Guanyuan (CV 4) and bilateral Zusanli (ST 36) (the first group) and bilateral Pishu (BL 20) and Weishu (BL 21) (the second group), and the two groups of acupoints were alternated every other day. The rats in the infrared group received infrared radiation on the stomach area and the area on the back between the T12-T13 spinous processes, the two areas were alternated every other day. Rats in the moxibustion group and the infrared group were treated for 20 min each time, once a day for 21 d. Rats in the blank group, the sham operation group, and the model group were simultaneously grasped and fixed, and no other treatment was performed. After 21 d of intervention, the rats in each group were fasted for 12 h, and blood was collected from the orbits. The numbers of CD3+, CD3+CD4+, CD3+CD8+ T lymphocytes in peripheral blood were determined by flow cytometry, and the ratio of CD3+CD4+/CD3+CD8+ was calculated. The rats were sacrificed and the thymus was dissected under sterile conditions to calculate the thymus index. Results:Compared with the blank group, the thymus index, peripheral blood CD3+, CD3+CD4+, CD3+CD8+ and CD3+CD4+/CD3+CD8+ ratio in the sham operation group did not change significantly (allP>0.05). Compared with the blank group, the thymus index of the model group was increased (P<0.05), the CD3+ and CD3+CD8+ T lymphocytes were increased (bothP<0.01), and the CD3+CD4+/CD3+CD8+ ratio was decreased (P<0.05). Compared with the model group, the thymus index of the moxibustion group was increased (P<0.01), and CD3+, CD3+CD4+ and CD3+CD8+ T lymphocytes and the ratio of CD3+CD4+/CD3+CD8+ in peripheral blood were increased (allP<0.05). Compared with the infrared group, the thymus index of the moxibustion group was significantly increased (P<0.05), the CD3+ and CD3+CD4+ T lymphocytes in the peripheral blood were significantly increased (bothP<0.01), and the CD3+CD8+ was increased (P<0.05). Conclusion:Moxibustion can significantly increase the thymus index of gastric cancer-bearing rats and activate CD3+CD4+ and CD3+CD8+ T lymphocytes in peripheral blood.
7.Effects of TNF-alpha receptor blocking peptide on adjuvant arthritis in rats.
Ya-ping HE ; Zhuo-ya LI ; Xiao-dan JIANG ; Wei FENG ; Yong XU ; Ping XIONG
Acta Pharmaceutica Sinica 2003;38(12):889-892
AIMTo study the effects of TNF receptor blocking peptide on adjuvant arthritis in rats.
METHODSThe model of rat adjuvant arthritis was induced by injection of complete Freund's adjuvant. The TNF receptor blocking peptide was injected locally in the ankle. The ankle swelling, the pathologic changes in the ankle joint and the expression of IL-1 beta mRNA and TNF-alpha mRNA by peritoneal macrophages (RT-PCR) were observed.
RESULTSThe model of rat adjuvant arthritis induced by injection of complete Freund's adjuvant was similar to human rheumatoid arthritis. The treatment with TNF receptor blocking peptide for 10 days resulted in complete inhibition of joint swelling, a decrease in infiltration of inflammatory cell into joint tissue, an obvious alleviation of inflammatory pathological damages and an apparent decline of TNF-alpha mRNA and IL-1 beta mRNA of peritoneal macrophages of rats.
CONCLUSIONThe TNF receptor blocking peptide can protect the joint from inflammatory damage induced by adjuvant arthritis by suppression of TNF-alpha and IL-1 production, thereby alleviating the pathological injury of joint and controlling effectively the clinic course of arthritis.
Animals ; Ankle Joint ; pathology ; Arthritis, Experimental ; immunology ; pathology ; Interleukin-1 ; biosynthesis ; genetics ; Macrophages, Peritoneal ; metabolism ; Male ; Peptides ; pharmacology ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Receptors, Tumor Necrosis Factor ; antagonists & inhibitors ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics
9.Study on expression of estrogen receptor isoforms in eutopic and ectopic endometrium of ovarian endometriosis.
Ai-jun LIU ; Zheng GUAN ; Zhuo-mei ZHANG ; Li-xin WEI ; Ya-li LI
Chinese Journal of Pathology 2008;37(9):584-588
OBJECTIVETo investigate the distribution of ER isoforms in endometriosis and eutopic endometrium.
METHODSTissue samples of patients with ovarian endometriosis, treated in People's Liberation Army General Hospital from January 2004 to December 2006, were retrieved. A total of 60 cases of ovarian endometriotic cysts with their corresponding eutopic endometrium (30 cases of proliferation phase and 30 of secretary phase eutopic endometrium) and 30 cases of normal endometrium (15 proliferative and 15 secretary phase endometrial samples respectively) were included. Expressions of ERalpha and ERbeta were analyzed using immunohistochemistry and the expression ratio was statistically analyzed by using SPSS 12.0 software.
RESULTSExpressions of both ERalpha and ERbeta in epithelial cells were positively correlated with that of the stromal cells. The expression of ERalpha in eutopic endometrium (73.3% in epithelium and 76.7% in stroma) was significantly higher than that in ovarian endometriotic cysts (43.3% in epithelium and 46.7% in stroma), or normal control (56.7% in epithelium and 50.0% in stroma, respectively, each P < 0.05. However, the expression of ERbeta (90.0% in epithelium and 76.7% in stroma) was higher in ovarian endometriotic cysts than that in the eutopic endometrium (68.0% in epithelium and 63.3% in stroma respectively, P < 0.05), and ERbeta expression in eutopic endometrium was higher than that in the normal control endometrium (36.7% in epithelium and 26.7% in stroma, respectively, P < 0.05). The expressions of both ERalpha and ERbeta changed periodically in eutopic and normal endometrium, whereas ERalpha and ERbeta level were less variable in the ectopic endometrium. The expression of ERbeta was statistically higher than that of ERalpha (P < 0.05) in ectopic endometrium, whereas no significant difference was seen between the two isoforms in the eutopic or normal endometrium.
CONCLUSIONSBoth ERalpha and ERbeta have higher expression levels in eutopic endometrium of patients with ovarian endometriotic cysts. ERbeta is predominantly expressed in endometriotic cysts, where the expression of ERalpha is limited. The different distribution of ERalpha and ERbeta may play an important role in the development of ovarian endometriosis.
Adult ; Choristoma ; pathology ; Endometriosis ; metabolism ; pathology ; Endometrium ; metabolism ; pathology ; Epithelium ; Estrogen Receptor alpha ; analysis ; Estrogen Receptor beta ; analysis ; Female ; Humans ; Immunohistochemistry ; Middle Aged ; Protein Isoforms ; analysis ; Receptors, Estrogen ; analysis ; Stromal Cells ; metabolism
10.Clinical application of flexible bronchoscopy and balloon dilatation in pediatric respiratory diseases under local anesthesia.
Wei GUO ; Li-Ya WAN ; Yong-Sheng XU ; Li-Xin REN ; Han-Quan DONG ; Zhuo FU
Chinese Journal of Contemporary Pediatrics 2012;14(11):859-862
OBJECTIVETo study the significance and safety of flexible bronchoscopy and balloon dilatation in the diagnosis and treatment of respiratory diseases in children.
METHODSA total of 438 children (236 males and 202 females) with respiratory diseases who were aged from 17 days to 15 years, were examined and/or treated by bronchoscopy (including bronchoscopic intervention) under local anesthesia.
RESULTSOf the 438 children, 311 were diagnosed with pulmonary infection, 68 with atelectasis, 36 with recurrent cough and asthma, 6 with hemoptysis of unknown origin, 6 with bronchial foreign body, 5 with congenital bronchopulmonary dysplasia, 2 with bronchiectasis, 1 with ciliary dyskinesia syndrome, 1 with lung tumor, and 2 with congenital immunodeficiency disease. After bronchoscopic examination, local flushing or bronchoalveolar lavage, and foreign body extraction, marked response was seen in 379 cases and response was seen in 46 cases. High-pressure balloon dilatation under bronchoscopy was performed in 5 cases with inflammatory stricture and achieved satisfying clinical effect. No severe complications were found in bronchoscopy.
CONCLUSIONSBronchoscopy and balloon dilatation under local anesthesia is safe and effective for the diagnosis and treatment of respiratory diseases in children.
Adolescent ; Anesthesia, Local ; Bronchoscopy ; adverse effects ; methods ; Child ; Child, Preschool ; Dilatation ; methods ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Respiratory Tract Diseases ; diagnosis ; therapy