1.Clinical application value of CT guided percutaneous lung biopsy in patients with negative fiberoptic bronchoscopy
Lifeng ZHANG ; Quan SHA ; Shumiao ZHANG ; Yanhua ZHANG
Chinese Journal of Postgraduates of Medicine 2017;40(1):36-39
Objective To evaluate the clinical application value of CT guided percutaneous lung biopsy in patients with negative fiberoptic bronchoscopy. Methods The clinical data of 51 patients with central type lung lesions were retrospectively analyzed and all patients had a negative result of fiberoptic bronchoscopy. Then they underwent the CT guided percutaneous lung biopsy. The number of paracentesis, complication and pathology were recorded. The patients were divided into 3 groups according to the tumor size:2.5-4.5 cm group, 4.6-6.5 cm group and>6.5 cm or tumor reaching the segmental bronchus group. Results The tumor diameter of 51 patients was 2.5-10.8 cm, average 4.9 cm. The number of paracentesis was 86 times, and 43 cases (84.3%) obtained definite pathological diagnosis: small cell carcinoma of lung in 14 cases, lung squamous cell carcinoma in 11 cases, adenocarcinoma in 6 cases, neuroendocrine carcinoma in 1 case, adenosquamous carcinoma in 4 cases, sclerosing hemangioma in 2 cases, tuberculosis in 2 cases, inflammatory pseudotumor in 2 cases, and large cell carcinoma in 1 case. The incidence of complication was 47.1%(24/51), among which the total slight pneumothorax occurred in 10 cases, intraoperative and postoperative hemoptysis, sputum with blood in 6 cases, greater amounts pneumothorax in 5 cases (3 cases underwent the closed drainage of pleural cavity), puncture path errhysis in 2 cases, thoracic cavity hemorrhage and thoracic wall haematoma in 1 case. No patient died. The 2.5-4.5 cm group had 22 cases, 4.6-6.5 cm group had 17 cases and >6.5 cm or tumor reaching the segmental bronchus group had 12 cases, and there was no statistical difference in the rate of definite pathological diagnosis among 3 groups:81.8%(18/22), 14/17 and 11/12, P>0.05;and there was statistical difference in the incidence of complication among 3 group:63.6% (14/22), 7/17 and 2/12, P<0.05. Conclusions The preferred examination of central type lung lesions patients is fiberoptic bronchoscopy, but because of diversity of disease, the CT guided percutaneous lung biopsy in the patients with negative fiberoptic bronchoscopy is feasible, safe, with high positive rate of biopsy and lower risk.
2.Evaluation of immune responses of human T lymphocyte subsets to Mycobacterium tuberculosis lipid antigens
Liang JIN ; Xi ZHANG ; Baiqing LI ; Quan SHA
Chinese Journal of Immunology 2016;(2):159-164
Objective:Some antigens of M.tb to culture with peripheral blood mononuclear cells ( PBMC) for assaying their proliferation and activation,so as to signify whether lipid antigens of M.tb have specific immune responses in host against M.tb infection or not.Methods:We treated PBMC with several lipid antigens of M.tb to explore the ability of these antigens to activate immunity in healthy individuals.We measured and analyzed cell proliferation by labeling cells with carboxyfluorescein succinimidyl amino ester (CFSE) and subjecting them to flow cytometry (FCM).The production of IFN-γ,TNF-αand IL-4 by T cell subsets (NKT,CD4+, CD8+,andγδT) from healthy donors was analyzed by FCM after stimulation with autologous immature dendritic cells pre-cultured with M.tb lipid antigens.The tested M.tb lipid antigens were the total lipid (TLIP),Acetone-Soluble Lipids (ASLIP),Purified Sulfolipid (PSLIP),Lipoarabinomannan (LAM) and Lipomannan (LM) levels.Medium free of lipid antigens(WCL,CFP,LPS,Mtb-HAg and blank) was used as a control.Results:We found the proportion of proliferative NKT and CD8+T cells significantly increased in all lipid groups (P<0.05).ASLIP,LAM and LM promoted non-proliferative CD4+T cells to secrete IL-4 and proliferative ones to secrete IFN-γ( P<0.05).All lipid antigens promoted both proliferativeγδT cells and CD8+T cells to secrete IFN-γand TNF-α,but the proportion of TNF-α-secreting cells in these populations decreased in the LM group ( P<0.05 ).Conclusion: Lipid antigens may affect the CD1-restricted T cells of the host to fight M.tb infection.
3.Advances in loop-mediated isothermal amplification in the diagnosis of tuberculosis
LUO Li-sha ; LIU Lin ; FENG Pin ; LAI Ji-jia ; CHEN Xue-yuan ; KONG Qing-quan
China Tropical Medicine 2022;22(11):1097-
Abstract: The loop-mediated isothermal amplification (LAMP) technique is a technique for the specific and efficient amplification of target fragments at a constant temperature using two pairs of specially designed primers and a strand displacement activity DNA polymerase. LAMP technique is a simple, rapid, specific, sensitive and cost-effective nucleic acid amplification method, and therefore has a promising future in the field rapid detection of Mycobacterium tuberculosis and grassroots applications. In this review, the basic principles and characteristics of the LAMP technique, the main molecular markers for the diagnosis of tuberculosis, and the use of different molecular markers and various types of novel techniques in the diagnosis of pulmonary tuberculosis, extrapulmonary tuberculosis, and drug-resistant tuberculosis were described. The LAMP technique has been widely used in the diagnosis of tuberculosis with high sensitivity and specificity, but the technique still has some shortcomings. This paper reviews the progress of its application in tuberculosis in recent years and provides an outlook on its development, with a view to providing a rational research direction for rapid diagnosis of tuberculosis in a resource-limited environment.
4.Signaling transduction pathways involved in basophil adhesion and histamine release
Quan SHA ; Poulsen K. LARS ; Gerwien JENS ; Nielsфdum ; Skov Stahl PER
Chinese Medical Journal 2006;19(2):122-130
Background Little is known about basophil with respect to the different signaling transduction pathways involved in spontaneous, cytokine or anti-IgE induced adhesion and how this compares to IgE-dependent and IgE-independent mediator secretion. The purpose of the present study was to investigate the roles of β1 andβ2 integrins in basophil adhesion as well as hosphatidylinositol 3-kinase (PI3K), src-kinases and extracellular signal regulated kinase (ERK)1/2 in basophil adhesion and histamine release (HR). Methods Basophils (purity of 10%-50%) were preincubated with anti-CD29 or anti-CD18 blocking antibodies before used for adhesion study. Basophils were preincubated with the pharmacological inhibitors wortmannin, PP1, PD98059 before used for adhesion and HR study. Cell adherence to bovine serum albumin (BSA) or fibronectin (Fn) was monitored using cell associated histamine as a basophil marker and the histamine was measured by the glass fiber assay.Results Basophil spontaneous adhesion to Fn was inhibited by anti-CD29. Interleukin (IL)-3, granulocyte/macrophage colony stimulating factor (GM-CSF) induced adhesion to BSA was inhibited by anti-CD18. Wortmannin at 1 μmol/L and PP1 at 20 μmol/L strongly interfered with, whereas PD98059 at 50 μmol/L weakly inhibited basophil spontaneous adhesion to Fn. One μmol/L wortmannin strongly inhibited IL-3, IL-5, GM-CSF and anti-IgE induced adhesion to BSA. PP1 at 20 μmol/L partly inhibited anti-IgE induced adhesion. Fifty μmol/L PD98059 marginally inhibited IL-5, weakly inhibited anti-IgE, partly inhibited GM-CSF induced adhesion. Wortmannin, PP1 and PD98059 inhibited anti-IgE (1:100 or 1:1000) induced basophil HR in a dose dependent manner. They inhibited calcium ionophore A23187 (10 μmol/L, 5 μmol/L) induced basophil HR in a dose dependent manner, but to different extend with PP1 being the most efficient.Conclusions Basophil spontaneous adhesion to Fn is mediated by β1-integrins whereas cytokine induced adhesion to BSA is mediated by β2-integrins. PI3K, src-kinases and ERK1/2 play distinct signaling roles in basophil adhesion and HR. PI3K is the key player while ERK1/2 is the weakest participant.
5.Lipoprotein glomerulopathy: a case report.
Hong-ling HAN ; Shan LIN ; Lin SONG ; Li-sha SUN ; Hui LIANG ; Li-quan ZHAO ; De-pei ZHAI
Chinese Journal of Pathology 2005;34(7):443-444
6.Inflammation activated bone marrow mesenchymal stem cell conditioned medium repairs radiation-induced acute injury to intestinal epithelial stem cells
Faxin MA ; Weihong SHA ; Qiyi WANG ; Jinliang LI ; Quan LU ; Yujun LUO
Chinese Journal of Tissue Engineering Research 2019;23(12):1324-1329
BACKGROUND: Our previous findings indicate that inflammation-activated bone marrow mesenchymal stem cell conditioned medium (MSC-CM) contribute to repairing the structure and function of the small intestine after radiation-induced acute intestinal injury. However, it is unclear whether the repair effect can be achieved by regulating small intestinal stem cells. OBJECTIVE: To investigate the effects of inflammation-activated bone marrow MSC-CM on the small intestinal epithelial stem cells after acute radiation-induced intestinal injury and to further discuss the repairing mechanism. METHODS: Bone marrow mesenchymal stem cells of Sprague-Dawley rats were separated, cultured and identified. Then, the bone marrow mesenchymal stem cells were co-cultured with normal or radiation-induced IEC-6 cell lines in the Transwell system for 24 hours. Inflammation-activated bone marrow mesenchymal stem cells were cultured alone for 48 hours. Non-activated MSC-CM (MSC-CMNOR) and MSC-CM under radiation-induced inflammatory condition (MSC-CMIR) were collected. Adult Sprague-Dawley rats (provided by the Experimental Center of Sun Yat-Sen University North Campus) were randomly divided into four groups with 20 rats in each group: control group, radiation group, radiation+MSC-CMNOR group and radiation+MSC-CMIR group. The rats in the latter three groups were exposed to one-off 14 Gy whole abdominal radiation to make a rat model of acute radiation-induced small intestinal injury. Three-day continuous administration beginning within 4 hours after successful modeling was given via the tail vein and intraperitoneal implantation of Alzet micro-osmotic pumps: EMEM-F12 (200 μL/d) for the radiation group, MSC-CMNOR for radiation+MSC-CMNOR group and MSC-CMIR for radiation+MSC-CMIR group. There was 2 mL of concentrated conditioned medium in the pump which was released at a constant rate of 10 μL/h into the abdominal cavity after implantation. Intestinal samples were collected at 1, 3, 5, 7 days after radiation for immunochemistry staining, western blot and qRT-PCR detection. RESULTS AND CONCLUSION: (1) On the 3rd day after radiation, Lgr5 positive cells, which were actively proliferating on the base of crypts, became significantly reduced compared with the normal control group, and there was nearly no existing Lgr5 positive cells. However, after infusion of MSC-CMIR, Lgr5 positive intestinal stem cells were significantly increased compared with the radiation group, while in the radiation+MSCNOR group, there was no significant increase in Lgr5 positive intestinal stem cells. (2) On the 3rd day after radiation injury, Bmi1 positive intestinal stem cells were almost invisible. After infusion of MSC-CMIR, Bmi1 positive intestinal stem cells increased significantly, and it was observed not only in the +4 cell position but also in the common position used to be Lgr5 stem cells, indicating that Bmi1 stem cells could differentiate into Lgr5 positive cells to act its repairing effect. (3) Western blot and qRT-PCR further confirmed that the radiation+MSC-CMIR group was significantly higher on the Lgr5 expression level than the radiation group and the radiation+MSC-CMNOR group, and it returned to the normal level on the 7th day after the continuous high expression level. The repair effect of radiation+MSC-CMNOR group was weaker, and only on the 7th day, the expression level of Lgr5 was statistically different from the radiation group. To conclude, inflammation-activated bone marrow MSC-CM exert a protective effect on the small intestinal epithelial stem cells after acute radiation-induced intestinal injury
7.Sesquiterpenes from stem of Schisandra glaucescens.
Pei-Pei ZHANG ; Sha-Sha GAO ; Tian-Tian ZHANG ; Jia-Chun CHEN ; Hong-Quan DUAN ; Jin-Bo FANG
China Journal of Chinese Materia Medica 2012;37(22):3426-3429
OBJECTIVETo study the chemical constituents from Schisandra glaucescens.
METHODThe chemical constituents were separated and purifed with silica gel, gel column chromatography preparative HPLC, and their structures were identified by such spectral methods as MS and NMR.
RESULTTwelve compounds were separated from petroleum ether fractions, and identified as t-cadinol (1), alpha-cadinol (2), torreyol (3), (+)-ent-epicubenol (4), ent-T-muurolol (5), (-)-15-hydroxycalamenene (6), (-)-cubebol (7), 4-epi-cubebol (8), caryophyllenol-I (9), caryophyllenol-II (10), oxyphyllenodiols A (11), caryolane-1,9/3-diol (12).
CONCLUSIONCompounds 4, 6-12 were separated from the genus for the first time, while compounds 1-12 were separated from this plant for the first time.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plant Stems ; chemistry ; Schisandra ; chemistry ; Sesquiterpenes ; chemistry ; isolation & purification
8.SMAD4 gene polymorphisms and genetic susceptibility of coal work's pneumoconiosis.
Jin XU ; Meng ZHU ; Hong-Quan CAI ; Kai ZHANG ; Wei-Wei DUAN ; Ting WANG ; Li WANG ; Sha-Sha WANG ; Yang YE ; Chun-Hui NI
Chinese Journal of Industrial Hygiene and Occupational Diseases 2010;28(10):766-771
OBJECTIVEto explore whether 6 tagging single nucleotide polymorphisms (SNPs) within SMAD4 gene were involved in the genetic susceptibility of coal worker's pneumoconiosis (CWP) by case-control study.
METHODSthis study consisted of 438 CWP patients and 448 controls. All study subjects were Han Chinese, underground coal miners and recruited from coal mines of Xuzhou Mining Business Group Co Ltd. The 5 ml venous blood sample was obtained from all studied subjects and extracted genome DNA from the isolated leucocytes. Six SNPs were selected from the HapMap and detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).
RESULTSthe single SNP analyses showed that the genotype frequencies of SMAD4 (rs10502913) was significantly different from those in controls (P < 0.05). Multivariate logistic regression analyses revealed that SMAD4 (rs10502913) AA genotype was associated with increased risk of CWP (adjusted OR = 1.63, 95%CI = 1.00 - 2.69, P = 0.05) and this was evident among subgroups of those smoker (adjusted OR = 2.28, 95%CI = 1.09 ∼ 4.80, P < 0.05) and cases with stage I (adjusted OR = 2.42, 95%CI = 1.41 ∼ 4.14, P < 0.01). The SMAD4 (rs9304407) GG genotype was associated with an decreased risk of CWP (adjusted OR = 0.65, 95%CI = 0.43 ∼ 0.98, P < 0.05) and the further stratification analysis showed that the risk of CWP was decreased in nonsmoking groups.
CONCLUSIONSour results suggest that individuals with the SMAD4 (rs10502913) AA genotype was associated with an increased risk of CWP. However, carriers of SMAD4 (rs9304407) GG genotype have a protective effect on the developing CWP.
Aged ; Anthracosis ; genetics ; Case-Control Studies ; Coal Mining ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Smad4 Protein ; genetics
9.Chemical constituents from stems of Hedyotis hedyotidea and their immunosuppressive activity.
Tian-tian ZHANG ; Sha-sha GAO ; Jun-jie HOU ; Yong-qin ZHOU ; Jie-wen ZHOU ; Xiao-gang WANG ; Nan QIN ; Jia-chun CHEN ; Hong-quan DUAN ; Jin-bo FANG
China Journal of Chinese Materia Medica 2015;40(12):2357-2362
Hedyotis hedyotidea has been traditionally used for the treatment of arthritis, cold, cough, gastro-enteritis, headstroke, etc. But few studies have screened the active compounds from extracts of H. hedyotidea. In this study, the structure of the chemical constituents from stems of H. hedyotidea were determined and the immunosuppressive activity of the compounds was evaluated. The compounds were separated and purified with silica gel, gel column chromatographies and preparative HPLC, and their structures were identified by spectral methods such as MS and NMR. Eleven compounds were obtained and identified as(6S,9S) -vomifoliol (1), betulonic acid (2), betulinic acid (3), betulin(4), 3-epi-betulinic acid (5), ursolic acid (6), β-sitosterol (7), stigmast-4-en-3-one (8), 7β-hydroxysitosterol (9), (3β,7β) -7-methoxystigmast-5-en-3-ol (10) and morindacin (11). This is the first report of compounds 1, 2, 4, 8, 9, 10 and 11 from H. hedyotidea. Compounds 1, 2 and 8-11 were firstly isolated from the genus Hedyotis, and compounds 9 and 10 were isolated from the family Rubiaceae for the first time. The immunosuppressive activity of these compounds was tested using the lymphocyte transsormationtest. Compounds 4, 6 and 9 showed significant immunosuppressive activity.
Animals
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Drugs, Chinese Herbal
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chemistry
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isolation & purification
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pharmacology
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Hedyotis
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chemistry
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Immunosuppressive Agents
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chemistry
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isolation & purification
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pharmacology
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Lymphocytes
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drug effects
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immunology
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Male
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Mass Spectrometry
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Mice
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Mice, Inbred C57BL
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Molecular Structure
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Plant Stems
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chemistry
10.Primary study on measuring the internal transcribed spacer I regions of rRNA genein seeds of Gentiana dahurica.
Ke-ping JI ; Xi-ling ZHANG ; Li-sha LIU ; Quan-yun LU ; Che CHENG
China Journal of Chinese Materia Medica 2003;28(4):313-316
OBJECTIVETo amplify the PCR with the internal transcribed spacerl regions measure the base sequence of the amplified products of DNA, and to set up an identified standard on the level of molecule.
METHODDNA from the seeds of G. dahurica was extracted by conventional method, and composed peculiar primer was used to amplify with the internal transcribed spacerl regions of the rRNA gene, and the base sequence of the amplified products by stopping the circle of the end of double deoxidation of four color fluorescent mark was measured.
RESULTIt was proved by agar sugar gel electrophoresis that the PCR amplified products of the internal transcribed spacerl regions of the rRNA gene existed. The base sequence of the seeds of G. dahurica's internal transcribed spacerl regions of the rRNA gene was measured.
CONCLUSIONTo measure the base sequence of internal transcribed spacerl regions of the rRNA gene in the seeds of G. dahurica's is a method to identify vegetal Chinese traditional medicine on the level of molecule.
Base Sequence ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Gentiana ; genetics ; Molecular Sequence Data ; Plants, Medicinal ; genetics ; Polymerase Chain Reaction ; RNA, Ribosomal ; genetics ; Seeds ; genetics