1.Long-term neurotoxic effect of intracephalic inflammation on substantia nigra dopaminergic neurons and the changes of asctrocytes in rats
Yongmei ZHAO ; Fengyue LV ; Qiuyan XU ; Ying YAN ; Qunyuan XU
Chinese Journal of Geriatrics 2010;29(5):416-419
Objective To investigate the role of asctrocytes in the process of chronic degeneration of dopaminergic neurons in intracephalic inflammation rat model induced by intracerebroventricularly injection of lipopolysaccharide.Methods Sixty healthy male SD rats were assigned into lipopolysaccharide group or saline control group randomly.All injections were made intracerebroventricularly on right side of the rats.Ethovison software was used to measure the movement distance of rats within 30 minutes.Specific antibody for glial fibrillary acid protein (GFAP) was used in immunohistochemistry stain to detect the changes of asctrocytes in the substantia nigra of rats.Results Movement distance of lipopolysaccharide-injected rats decreased by 21.2% compared with saline-injected rats at 16 weeks after injection (t=2.54,P<0.05)by 27.0% (t=3.55,P<0.01) at 24 weeks and by 31.4% (t=3.91,P<0.01) at 28 weeks after lipopolysaccharide injection.The asctrocytes were activated obviously in the substantia nigra of lipopolysaccharide-injected group at 2 weeks,while the numbers of GFAP-positively stained cells (228.60 + 22.35) increased significantly compared with saline-injected group ( 165.20 ± 25.97) (t = 4.14,P< 0.05).The activation of asctrocytes was not found in lipopolysaccharide-injected group at 4 weeks and 12 weeks.The asctrocytes were re-activated in the substantia nigra of lipopolysaccharide-injected group at 24 weeks,while the numbers of GFAP-positively stained cells (220.00±21.01 ) increased significantly compared with saline-injected group (169.00± 19.00) (t= 4.03,P<0.05).The activation of asctrocytes was not seen at any time point in saline-injected group.Conclusions Intracephalic inflammation induces chronic degeneration of substantia nigral dopaminergic neurons in rats.The asctrocytes exhibite "acute activation-quiescing-reactivation" state,indicating that they might be involved in the mechanism of dopaminergic neurons degeneration.
2.Mechanism of cyclin-dependent inhibitor p27~(Kip1) in regulating the differentiation of immortalized human neural progenitor cells
Yongmei ZHAO ; Qiuyan XU ; Weihong LI ; Qunyuan XU ; Haiyan ZHANG
Acta Anatomica Sinica 2010;41(1):22-26
Objective To investigate whether there is any functional link between p27~(Kip1) function and all-trans retinoic acid (RA) in the control of neuronal differentiation of immortalized human neural progenitor cells (hSN12W-TERT cells). To investigate the mechanism by which p27~(Kip1) regulates the differentiation of immortalized human neural progenitor cells. Methods hSN12W-TERT cells were derived from the striatums of human embryos at 12 weeks gestation and cultured with serum-free medium in presence of EGF and bFGF. At the appropriated time, hSN12W-TERT cells were exposed to 1μmol/L RA for 3, 5, 7 days respectively. The experiment was repeated there times. Cell cycle analysis was performed by flow cytometry analysis (FACS). The expression of p27~(Kip1), p21~(cip1), cyclin-dependent kinase 2 (cdk2), p-cdk2 and S-phase kinase-associated protein 2 (skp2) in hSN12W-TERT cells before and after RA treatment cells were determined by using Western blotting analysis. Results FACS result showed that 77.25% of proliferating hSN12W-TERT cells were in the G1/G0-phase while 9.38% of cells in the S-phase. Following RA treatment, cell growth was arrested, and 85.68% of cells accumulated in G1/G0-phase while 8.57% of cells in the S-phase. Western blotting analysis demonstrated that the levels of p27~(Kip1) in the hSN12W-TERT cells increased following 3 days' treatment with RA compared with those of normal untreated cells, with a peak at 5 days (P<0.05). The similar results were acquired both in nuclear proteins and in cytoplasm proteins of hSN12W-TERT cells. The expression level of p21~(cip1) decreased in response to RA treatment. RA did not affect the expression of cdk2, but the expression of p-cdk2, which represented the activity of cdk2, was markedly decreased in response to RA treatment. Skp2, which was required for the ubiquitin-mediated degradation of p27~(Kip1), was detected in proliferating hSN12W-TERT cells. The expression of skp2 reduced dramatically in response to RA treatment in a time-dependent manner.Conclusion There is a functional link between RA and p27~(Kip1) function in the control of neuronal differentiation in hSN12W-TERT cells. P27~(Kip1) plays a key role during neuronal differentiation. Moreover, high levels of p27~(Kip1) are associated with its degradation inhibiting through reducing proteasome-dependent proteolysis.
3.Impacts on asthma at persistent stage and immune function in the patients treated with acupuncture for warming yang and benefiting qi.
Yilin XIE ; Wenrong WAN ; Yinlong ZHAO ; Junjie XIE ; Qiuyan WU
Chinese Acupuncture & Moxibustion 2015;35(11):1089-1093
OBJECTIVETo compare the difference in clinical efficacy on bronchial asthma at chronic persistent stage between acupuncture for warming yang and benefiting qi and seretide.
METHODSOne hundred and eighty patients of bronchial asthma at chronic persistent stage were randomized into an acupuncture group and a western medication group, 90 cases in each one. In the acupuncture group, acupuncture for warming yang and benefiting qi was applied at Dazhui (GV 14), Feishu (BL 13), Danzhong (CV 17), Dingchuan (EX-B 1), Jianshi (PC 5), Zhigou (TE 6), Taixi (KI 3) and Zusanli (ST 36), once every two days. In the western medication group, inhalation therapy with seretide was applied, 1 to 2 inhalations each time, twice a day. The treatment for 20 days was as one session in the two groups, at the intervals of 2 days after each session. Four sessions of treatment were required. The immune function indices were observed before and after treatment in the patients of two groups, named immunoglobulin IgG, IgM and IgE; peripheral T lymphocytes (CD3+), helper T lymphocytes (CD4+), inhibitory T lymphocytes (CD8+) and the ratio of CD4+ and CD8+; as well as the pulmonary ventilation function indices, named maximum pulmonary expiratory flow (PEF), forced expiratory volume in one second (FEV1) and forced vital capacity (FVC). The clinical efficacy was compared between the two groups.
RESULTSThe total effective rate was 93.3% (84/90) in the acupuncture group, better than 88.9% (80/90) in the western medication group (P < 0.05). After treatment, the immune function indices and pulmonary ventilation function indices were apparently improved as compared with those before treatment in the two groups (all P < 0.05). The improvements in the acupuncture group were much more significant (all P < 0.05).
CONCLUSIONAcupuncture for warming yang and benefiting qi effectively controls the symptoms of bronchial asthma at chronic persistent stage and improves immune and pulmonary functions. The efficacy is better than that of seretide.
Acupuncture Points ; Acupuncture Therapy ; instrumentation ; methods ; Adolescent ; Adult ; Asthma ; immunology ; physiopathology ; therapy ; Female ; Forced Expiratory Volume ; Humans ; Immunoglobulins ; immunology ; Male ; Middle Aged ; Qi ; T-Lymphocytes ; immunology ; Treatment Outcome ; Young Adult
4.Role of MicroRNA-21 in Pathogenesis,Diagnosis and Treatment of Pancreatic Cancer
Qiuyan ZHAO ; Sumin CHEN ; Yingchun REN ; Baiwen LI
Chinese Journal of Gastroenterology 2017;22(8):502-505
Pancreatic cancer is characterized by high invasiveness,high malignancy and poor prognosis. Therefore,it is urgent to find biomarkers for early diagnosis and to develop targeting drugs for treatment of pancreatic cancer. MicroRNAs (miRNAs)are single-stranded,non-coding small RNA molecules with a length of 18-25 nucleotides and are highly expressed in pancreatic cancer. MiRNAs play important roles in cell proliferation,differentiation,apoptosis,invasion and metastasis,and are associated with adverse prognosis and resistance to chemotherapy in pancreatic cancer. As a new target for diagnosis and treatment of pancreatic cancer,miRNA-21 has become the hot spot of clinical research. This article reviewed the advances in studies on miRNA-21 in the pathogenesis,diagnosis and treatment of pancreatic cancer.
5.Regulation of prostaglandin E_2 on orphan nuclear receptor Nurr1 expression in MN9D cells
Yongmei ZHAO ; Haiyan ZHANG ; Qiuyan XU ; Fengyue LU
Chinese Pharmacological Bulletin 1987;0(01):-
Aim To investigate whether prostaglandin E2(PGE2)can increase endogenous Nurr1 expression in a dopamine-synthesizing cell line(MN9D) with immature characteristics and to study whether tyrosine hydroxylase(TH) expression is up-regulated in response to Nurr1-overexpression,in order to investigate the role of Nurr1 during dopaninergic neurons development.Methods MN9D cells were treated with 100 ?g?L-1 PGE2 for 2 h to 6 h.The morphology changes of MN9D cells were observed under phase-contrast microscope. The expression of Nurr1 and TH in MN9D cells was analyzed by using immunohistochemical staining and Western blot analysis.Results ① The morphology of MN9D cells did not change significantly following PGE2 treatment for 2 h,4 h and 6h compared with that of MN9D cells left untreated.② Nurr1-positive staining in MN9D cells treated with PGE2 for 2 h,4 h and 6 h was much stronger than that of untreated cells while the percent of TH-positive MN9D cells after PGE2 treatment for 2 h,4 h and 6 h was similar to that of untreated.③ The expression of Nurr1 protein in MN9D cells treatment with PGE2 for 6 h was significantly higher than that of untreated(P
6.Clinical study of clearing liver fire, removing dampness, strengthening slpeen and tonifying kidney methods in treating middle-aged and old woman with chronic urinary tract infection
Kaisheng ZHAO ; Baoli LIU ; Wei WEI ; Qiuyan MA ; Wenjing ZHAO ; Shengrong ZHANG
International Journal of Traditional Chinese Medicine 2011;33(11):976-978
ObjectiveTo observe the clinical effect of the treatment of clearing liver fire,removing dampness,strengthening spleen and tonifying kidney in treating middle-aged and old woman with chronic urinary tract infection.Methods82 cases were randomly divided into two groups.42 cases in treatment group were treated with herbal prescription of clearing liver fire,removing dampness,strengthening spleen and tonifying kidney,and 40 cases in control group were treated with sensitive antibiotics.The clinical effect,safety and changes of immune index were compared between two groups.ResultsIn the treatment group,the recent cure rate was 76.8%,the total effective rate was 90.5% and the complete cure rate was 61.9%; while in the control group,the recent cute rate was 50%,the total effective rate 65% and the complete cure rate 27.5%.It showed significant difference between two groups (Z=2.740,P<0.05).There was no significant difference between two groups in hepatic and renal function,serum and urine β2-MG level before and after treatment and it showed a better clinical safety in treatment group.The level of IgA,IgG and IgM increased after the treatment in treatment group.Comparing with the control group,the results showed significant differences(t=9.1,10.5 and 11.2 respectively,P<0.01).In treatment group the level of CD4+was increased,CD8+was decreased,and the ratio of CD4+/CD8+ was increased obviously after the treatment.The differences before and after the treatment and between groups were significant(t=13.3,8.6,P<0.01).In control group the level of CD4+、CD8+、CD4+/CD8 +did not change significantly.The differences before and after the treatment had no statistical significance (P>0.05).ConclusionThe treatment with herbs of clearing liver fire,removing dampness,strengthening slpeen and tonifying kidney had better effect,less side effect and good research prospect.
7.Progress in establishment and application of feline calicivirus reverse genetics operating system.
Yanli ZHAO ; Hongwei DONG ; Xiaoqing CHEN ; Chao GAO ; Qiuyan LIU ; Songtao YANG ; Guixue HU
Chinese Journal of Virology 2015;31(1):74-79
Feline calicivirus (FCV) is an important and highly prevalent pathogen of cats that causes feline respiratory disease. The reverse genetic systems for FCV have been established in national and international laboratories since 1995. This technique has been used widely in FCV basic research and good progress has consequently been made to determine the relationship between viral genome structures and the function of their proteins, the expression of foreign proteins, virus-host interactions, and viral pathogenic mechanisms. In this article,we review the state of progress with regards to the establishment and application of the FCV reverse genetic operating system,which will provide a useful reference tool for future related research.
Animals
;
Caliciviridae Infections
;
veterinary
;
virology
;
Calicivirus, Feline
;
genetics
;
metabolism
;
Cat Diseases
;
virology
;
Cats
;
Reverse Genetics
;
methods
;
trends
;
Viral Proteins
;
genetics
;
metabolism
8.Cytogenetic and molecular genetics of a rare case with Turner syndrome
Junke XIA ; Yanxia LIU ; Yongjiang ZHAO ; Yaqin HOU ; Ning LU ; Qiuyan ZHANG ; Xiangdong KONG
Chinese Journal of Endocrinology and Metabolism 2021;37(2):162-164
Turnner syndrome is a common sex chromosome disorder. We reported a rare case with Turnner syndrome caused by abnormal number and structure of sex chromosomes. Hereby fluorescence in situ hybridization (FISH) and copy number variation by whole genome low depth sequencing (CNV-seq) were used to clarify the abnormal chromosome. This study provides a diagnostic strategy for clinicians and genetic researchers.
9. Role of MicroRNA-21 in Proliferation and Migration of Pancreatic Cancer Cells
Chinese Journal of Gastroenterology 2019;24(10):586-590
Background: Studies have shown that microRNAs are closely related to the occurrence and development of tumors. As one of the most common carcinogenic microRNAs, the specific role and mechanism of miR-21 in the proliferation and migration of pancreatic cancer cells have not been fully elucidated. Aims: To investigate the effect and specific molecular mechanism of miR-21 on proliferation and migration of pancreatic cancer cells. Methods: PANC-1 and MIA PaCa-2 cells were transfected with lentiviral vectors to construct stable cell lines with overexpression and knockdown of miR-21, respectively. The transfection efficiency was verified by qRT-PCR. CCK-8 assay was used to detect cell proliferation, scratch test was used to detect cell migration ability. The expressions of Spry2 mRNA and protein were determined by qRT-PCR and Western blotting, respectively, and luciferase report assay was used to detect the regulatory effect of miR-21 on Spry2. Results: Overexpression of miR-21 significantly promoted PANC-1 cells proliferation and migration (P<0.05). Knockdown of miR-21 significantly inhibited MIA PaCa-2 cells proliferation and migration (P<0.05). Compared with controls, expressions of Spry2 mRNA and protein were significantly decreased after up-regulation of miR-21 (P<0.05), however, expressions of Spry2 mRNA and protein were significantly increased after down-regulation of miR-21 (P<0.05). Overexpression of miR-21 could inhibit luciferase activity of plasmids containing wild-type Spry2 3'-UTR sequence (P<0.05), and knockdown of miR-21 could enhance luciferase activity (P<0.05). Spry2 could reverse miR-21-mediated cell proliferation and migration (P<0.05). Conclusions: MiR-21 can affect biological function of pancreatic cancer cells by targeted regulating the expression of Spry2.
10.Lymphogranuloma venereum complicated by squamous cell carcinoma: a case report
Quanzhong LIU ; Yan LI ; Qiuyan YANG ; Mei WANG ; Weifeng YAO ; Liyuan ZHAO ; Yong LIU ; Suying ZHANG ; Zhiyi FU
Chinese Journal of Dermatology 2011;44(8):567-570
A 53-year-old man was admitted to the hospital for verrucous hyperplasia on the circular sulcus for 2 years as well as erythematous painful swelling, ulcer and rupture of inguinal lymph nodes for more than 2 months. Physical examination revealed erythematous, indurated and painful swelling of bilateral inguinal lymph nodes. Fluctuation could be felt at the centre of the right swollen inguinal lymph nodes, where several pores were seen with yellowish-white purulent fluid flowing out, giving the appearance of a watering can. Multiple swollen or ulcerative lymph nodes were separated by the inguinal ligament forming the groove sign.There were irregular, indurated, verrucous, proliferative and keratinized lesions sized 1 cm × 2 cm on both sides of the circular sulcus. The right scrotum was obviously swelling. Erythematous, fluctuating swelling, ulcer and rupture of the glans fraenum were also observed with yellowish-white purulent exudates. Neither the secretion from the skin lesions on the surface of bilateral inguinal lymph nodes nor the puncture sample from the right groin was positive for multiple fungal or bacterial culture, acid-fast stain or first culture of C. trachomatis.However, the endogenous plasmid of Chlamydia was successfully amplified by PCR from these samples, and restriction fragment length polymorphism (RFLP) analysis of the major outer membrane protein (MOMP) suggested that the genotype of the Chlamydia strain was L3. Western blot revealed the presence of anti-MOMP antibodies and anti-Pmp H antibodies (titer: > 1: 800) in serum. Culture of C. trachomatis also gave positive results after multiple passage. Biopsy of the verrucous hyperplasia on the right groin is consistent with well-differentiated squamous cell carcinoma of the skin. A diagnosis of lymphogranuloma venereum complicated by cutaneous squamous cell carcinoma was made.