Background Most animal models of experimental autoimmune uveitis (EAU) are single attacked procedure,with a different feature from the natural course of human recurrent autoimmune uveitis.So establishing a recurrent EAU model is necessary for the clinical study on EAU.Objective This study was to establish the recurrent EAU model in rat and investigate the ocular inflammation and pathological manifestation and interleukin-17 (IL-17)expression in the eye.Methods T cells isolated from the spleen and draining lymph nodes of Lewis rats immunized with interphotoreceptor retinoid-binding protein (IRBP) 1177-1191 peptide fragments (R16) 10 days earlier were re-stimulated with R16 in vitro and injected into naive syngeneic rats to establish the recurrent EAU models,and the normal Lewis rats were used as controls.The eyes of model rats were then examined daily for clinical signs of uveitis by slit-lamp biomicroscopy and scored Caspi's criteria.The rats were sacrificed 1 month,2,3months after injection respectively,and the retinal sections were prepared for the pathological examination by hemotoxylin & eosin staining.Immunohistochemistry was performed to detect the expression of IL-17 in the retina.Results Adoptive transfer of R16-specific T cells to Lewis rats induced recurrent uveitis.The inflammatory scores on the fourth day,the sixth day,and the inflammatory response disappeared on the tenth day after injection.However,the inflammatory reaction occurred repeatedly 4 or 5 times in the 2-month duration after that,and the right and left eyes of a single recipient showed a different pattern of relapse,and the clinical manifestations of EAU was similar to the natural course to those of human autoimmune uveitis.In the retinal specimens of 1-,2-and 3-month group,the number of inflammatory cells was gradually decreased as the time lapse.Compared with the normal group,the thicknesses of the entire retina,outer nuclear layer and inner nuclear layer decreased with a significant difference among the 4 groups (F=20.46,288.40,4.43,all P=0.00).The number of RGCs in the normal group,1-,2-and 3-month group was 231.27 ± 15.36,225.36 ± 17.79,132.18 ±9.39 and 67.45 ± 11.90,respectively,showing a significant difference among them (F=68.94,P=0.00).Immunohistochemistry showed that the scores of the IL-17 expression in the rat retina were 0.64 ± 0.17,1.92 ± 0.19,1.17 ± 0.23 and 0.83 ± 0.23,showing statistically significant difference (F=64.10,P=0.00).Conclusions The stimulation of R16-specific T cells can induce recurrent EAU in Lewis rat.Th17 is involved in the disease course.

OBJECTIVETo observe the effect of Yangjing Zhongyu Decoction (YZD) on mRNA and protein expression of PCNA, StAR, and FSHR in ovarian granulose cells (GCs) cultured by excess androgen.

METHODSOvarian GCs from porcine follicles were isolated and cultured in vitro. Follicular stimulating hormone (FSH) or YZD was added in the GCs treated by excess testosterone propionate. Totally 48 h later mRNA and protein expression of PCNA, StAR, and FSHR were detected by RT-PCR and Western blot.

RESULTSExcess androgen inhibited mRNA and protein expression of PCNA, StAR, and FSHR of GCs. FSH and YZD could antagonize inhibition of excess androgens, and promote mRNA and protein expression of PCNA, StAR, and FSHR in GCs.

CONCLUSIONYZD could antagonize the inhibition of excess androgen on mRNA and protein expression of PCNA, StAR and FSHR in GCs. Thus, we inferred that YZD could improve the follicle dysplasia by promoting mRNA and protein expression of PCNA, StAR and FSHR in GCs.

Androgens ; pharmacology ; Animals ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Female ; Follicle Stimulating Hormone ; pharmacology ; Granulosa Cells ; cytology ; drug effects ; metabolism ; Membrane Transport Proteins ; genetics ; metabolism ; Ovarian Follicle ; cytology ; drug effects ; Proliferating Cell Nuclear Antigen ; genetics ; metabolism ; RNA, Messenger ; genetics ; Receptors, FSH ; genetics ; metabolism ; Swine

Adult ; Aged ; Aged, 80 and over ; Epistaxis ; therapy ; Female ; Gloves, Protective ; Hemostatic Techniques ; Humans ; Male ; Middle Aged

Objective:To explore the relationship between electrocardiogram QRS wave duration and pathological parameters of acute myocardial infarction and its prognosis.Methods:The patients with acute myocardial infarction treated in Hunan Brain Hospital from January 2018 to June 2019 were divided into normal group (≤110 ms, NG group) and extended group (>110 ms, EG group) according to the time limit of QRS wave group of electrocardiogram (ECG). The clinicopathological data of the two groups were collected, and the differences of relevant pathological parameters between the two groups were compared. The correlation between ECG QRS complex time limit and pathological parameters of acute myocardial infarction and the independent risk factors affecting the occurrence of malignant cardiovascular events after acute myocardial infarction were analyzed.Results:A total of 134 patients were enrolled, 73 in the EG group, and 61 in the NG group. The plasma levels of N-terminal pro-brain natriuretic peptide (N-proBNP) in EG group were significantly higher than those in NG group ( P<0.05), and left ventricular ejection fraction (LVEF) was significantly lower than that in NG group ( P<0.05). The time limit of QRS wave group in EG group was positively correlated with the plasma levels of N-proBNP ( r=0.981, P<0.001) and negatively correlated with LVEF ( r=-0.979, P<0.001). The ratio of Killip grade Ⅰ patients in EG group were lower than those in NG group ( P<0.05), while the ratio of grade Ⅱ and Ⅲ patients were significantly higher than those in NG group ( P<0.05), and there was no significant difference between the two groups in grade Ⅳ patients ( P>0.05). The incidence of malignant cardiovascular events (cardiogenic shock, malignant arrhythmia, acute pulmonary edema, cardiogenic death and re-infarction) in EG group was significantly higher than that in NG group ( P<0.05). Multivariate logistic regression analysis showed that elevated serum N-proBNP, decreased LVEF, prolonged QRS wave duration and high Killip cardiac function were independent risk factors for malignant cardiovascular events after acute myocardial infarction. Conclusions:After acute myocardial infarction, EG patients have worse cardiac function, higher incidence of malignant cardiovascular events and worse prognosis than NG patients.

OBJECTIVETo construct an oncolytic adenovirus with the DD3 promoter regulation, expressing small hairpin RNA and targeting the SATB1 gene (SATBI-shRNA), and to evaluate its potential for inhibiting the growth of human prostatic carcinoma cells (LNCaP) in vitro.

METHODSSATB1-shRNA expression cassettes containing the H1 promoter were produced by PCR from pSilencer3. 1-SATB1 and inserted into the pZD55 vector, and the recombinant plasmid pZD55-SATB1-shRNA was constructed, pZD55SATB1-shRNA and pZXC2-DD3-E1A were digested with EcoRV and Xba I , and the obtained expression cassettes linked each other to construct the recombinant plasmid pDD3-ZD55-SATB1, which was cotransfected with the pBHGE3 packaging plasmids mixture into 293 cells by Effectence. The recombined adenoviruses DD3-ZD55-SATB1 were identified by PCR. Viruses were propagated on HEK293 cells and purified by standard techniques, and the functional PFU titers determined by plaque assay on 293 cells. The antitumor potential of DD3-ZD55-SATB1 to LNCaP was evaluated by the crystal violet dye method. The expression level of the E1A gene was detected by Western blot, and that of the SATB1 gene in the adenovirus-infected LNCaP cells by both Western blot and RT-PCR.

RESULTSAn oncolytic adenovirus expressing SATB1-shRNA with the DD3 promoter regulation, DD3-ZD55-SATB1, was constructed successfully, whose functional PFU titer was 1.2 x 10(10) PFU/ml. DD3-ZD55-SATB1 showed an obvious cytopathic effect and a selective expression of E1A in the adenovirus-infected LNCaP cells; it exhibited a high LNCaP-targetability and specific SATB1-silencing effect.

CONCLUSIONThe successful construction of the oncolytic adenovirus DD3-ZD55-SATB1 offers a new tool for researches on the gene therapy for human prostate cancer.

Adenoviridae ; genetics ; Carcinoma ; therapy ; Cell Line, Tumor ; Genetic Vectors ; Humans ; Male ; Matrix Attachment Region Binding Proteins ; genetics ; Oncolytic Virotherapy ; methods ; Oncolytic Viruses ; genetics ; Promoter Regions, Genetic ; Prostatic Neoplasms ; therapy ; RNA Interference ; RNA, Small Interfering ; genetics

Sodium calcium exchanger (NCX), which is widely expressed in the plasma membrane, mitochondrial membrane and secretory vesicles in diverse kinds of cells, belongs to a type of cation translocators. NCX works in two modes, the forward mode and reverse mode, to regulate the intracellular Ca(2+) concentration bi-directionally. In the forward mode, NCX carries Ca(2+) out of the cell against its electrochemical gradients coupled to the influx of Na(+) down its electrochemical gradients; alternatively, Ca(2+) enters through the reverse mode of NCX, and Na(+) is carried out of the cell. Exactly through the two-way modes, NCX can regulate intracellular Ca(2+) concentration fleetly and accurately, and plays a critical role in a series of physiological processes including intracellular signal transduction, growth and development of cells, excitation and its coupled functions of excitable cells. NCX are acknowledged to be involved in myofiber contraction, neurotransmission, migration and differentiation of neurogliocyte, activation of immune cells, secretion of cytokines and hormones etc. Moreover, abnormal activation of the reverse mode of NCX plays a vital role in many pathological processes including cell apoptosis, ischemia-reperfusion injury, insulin secretion, tumor etc. Here we reviewed the research status about the NCX's participation in some physiological and pathophysiological processes, so as to provide comprehensive understanding about its functions.
Animals ; Apoptosis ; Calcium ; physiology ; Humans ; Ion Transport ; Reperfusion Injury ; physiopathology ; Signal Transduction ; Sodium ; physiology ; Sodium-Calcium Exchanger ; physiology

OBJECTIVESTo further discuss the method of evaluating curative effect on erectile dysfunction(ED).

METHODSClinical trials(Phase II) of sildenafil and phentolamine were both 8-week-long, randomized, double-blind and placebo-controlled. Not only the curative effect between each treatment group and placebo group but also between the treatment groups were compared. The dosage of sildenafil and phentolamine was respectively 50-100 mg and 40 mg.

RESULTSEfficiency, success rate of sexual intercourse and general curative effect in sildenafil and phentolamine groups were respectively 79.17%, 75.00%, 83.33% and 52.38%, 85.71%, 52.38%. The result revealed a statistically significant (P < 0.05) improvement over placebo. But no obvious differences were shown between the two treatment groups(P > 0.05).

CONCLUSIONSMost assessments of curative effect of ED drugs come from questionnaires answered subjectively by patients. The Lack of objective assessment criteria way lead to the non-conformity between trial results and clinical practices. So the curative effect of drugs shall be assessed more accurately and from more perspectives.

Adrenergic alpha-Antagonists ; therapeutic use ; Double-Blind Method ; Erectile Dysfunction ; drug therapy ; Humans ; Male ; Phentolamine ; therapeutic use ; Piperazines ; therapeutic use ; Purines ; Sildenafil Citrate ; Sulfones ; Surveys and Questionnaires ; Treatment Outcome ; Vasodilator Agents ; therapeutic use

OBJECTIVETo explore the relationship between hypoxia and the apoptosis of corpus cavernosum smooth muscle cells (CCSMC) in SD rats.

METHODSCCSMCs were cultured in vitro and identified by immunohistochemistry, and then underwent hypoxia interference at the concentration of 1% O2 for 12, 24, 48 and 72 hours, with normal oxygen concentration as the control. Flow cytometry was used to determine the cycles and apoptosis of the cells.

RESULTSThe cultured CCSMCs grew well, positive for anti-smooth muscle alpha-actin monoclonal antibody immunohistochemical staining. Flow cytometry showed that the number of CCSMCs in G0/G1 was gradually increased within 48 hours and then decreased, just opposite to the proportion of the S phase cells. But no regular change was found in the proportion of the cells in the G2/M phase.

CONCLUSIONHypoxia promotes the apoptosis of CCSMCs in a time-dependent manner, to the maximum at 48 hours, and then cell lysis may occur, but with no further apoptosis.

Animals ; Apoptosis ; Cell Hypoxia ; Cells, Cultured ; Male ; Muscle, Smooth, Vascular ; cytology ; Penis ; pathology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo compare the effects on lumbar disc snake herniation between acupuncture in combination of snake moxibustion with simple acupuncture or snake moxibustion.

METHODSOne hundred and twenty cases of lumbar disc herniation patients were randomly divided into an acupuncture and snake moxibustion group (group A, 60 cases), a acupuncture control group (group B, 30 cases) and a snake moxibustion control group (group C, 30 cases). In group A, acupuncture was applied to Shenshu (BL 23), Dachangshu (BL 25) and tender points bilateral to the spine, once a day. Additionally, at the end of acupuncture on the 1st day of a week, snake moxibustion was applied for one time. In group B and group C, acupuncture or snake moxibustion was applied respectively.

RESULTSThe excellent rate was 95.0% (57/60) in group A, which was superior to that of 70.0% (21/30) in group B and 66.7% (20/30) in group C (both P < 0.05). JOA scores in each group after treatment increased apparently (all P < 0.01), of which, the scores in group A increased much more apparently as compared with those of the other two groups after treatment (both P < 0.01). The efficacy in group A was relevant obviously with the duration of sickness, the shorter the duration of sickness was, the higher the excellent rate would be. The age of patients in group A was relevant obviously with the efficacy, the younger patient was, the higher excellent rate would be.

CONCLUSIONThe efficiency on lumbar disc hernation treated with acupuncture and snake moxibustion is superior to that with either acupuncture or snake moxibustion.

Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Animals ; Female ; Humans ; Intervertebral Disc Displacement ; physiopathology ; therapy ; Lumbar Vertebrae ; physiopathology ; Male ; Middle Aged ; Moxibustion ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the effect of adenovirus expressing human bone morphogenetic protein-7 (hBMP-7) on proliferation and differentiation of human dental pulp cells.

METHODSThe replication-deficient adenoviral vector encoding hBMP-7 gene was constructed by using homologous recombinant modality. The efficiency of transfection was evaluated by fluorescent microscopy and flow cytometry. The expression of hBMP-7 protein in adenovirus-infected dental pulp cells was determined by Western blot. The proliferation of cells was tested by MTT method, the activity of alkaline phosphatase was assayed, von Kossa staining was used to detect mineralized nodule formation, and the expression of DSPPmRNA in cells was detected using semi-quantitative RT-PCR.

RESULTSGreen fluorescent protein was visible under fluorescent microscopy. Higher transfection efficiency (91.1 +/- 1.0)% could be obtained at MOI of 75. Western blot from dental pulp cells infected with Ad-hBMP-7 for 48h detected protein expression of a hBMP-7 gene. The activity of alkaline phosphatase in cells was significantly higher than those of the control groups (P < 0.05). The cells infected with Ad-hBMP-7 had the ability of mineralization. DSPP mRNA expression of cells was in a time- and dose- dependent manner.

CONCLUSIONSAd-hBMP-7 can induce human pulp cells into odontoblasts, but has no obvious effect on their proliferation.

Adenoviridae ; genetics ; Bone Morphogenetic Protein 7 ; genetics ; metabolism ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Dental Pulp ; cytology ; Humans ; Odontoblasts ; cytology ; Transfection