Apoptosis ; Down-Regulation ; Electron Transport Complex IV ; chemistry ; genetics ; metabolism ; Humans ; Mitochondria ; metabolism ; Mutation ; Neoplasms ; genetics ; metabolism ; pathology

OBJECTIVETo study the effects of Citalopram on the mRNA expression of bax and bel-2 in frontal cortical neurons and on cell apoptosis of rats after stress.

METHODSTwenty-four healthy male SD rats were randomly divided into three groups (n = 8). The control group did no receive any treatment, the stress group was subject to stress and given normal saline and experimental group was given Citalopram irrigation stomach after stress. Rats were forced to swim to establish chronic stress model (15 min/d, 4 weeks), bax, bcl-2 mRNA expression were tested by in situ hybridization technique (ISH), TUNEL assay was used to determine cell apoptosis, Nikon image analysis software were used to measure the number of positive cells in each index.

RESULTSCompared with the control group, the stress group showed a larger number of bax mRNA expressing cells( P < 0.01), a smaller number of bcl-2 mRNA expressing cells (P < 0.01), and the staining intensity of positive cells was significantly reduced( P < 0.01). Compared with the stress group, the experiment group showed more reduced number of bax mRNA positive cells( P < 0.01) and significantly increased bcl-2 mRNA positive cells( P < 0.05), a small amount of positive cells were found, compared with that in the stress group, nuclear condensation in the experimental group was reduced significantly and the staining was obviously weaker( P < 0.01).

CONCLUSIONCitalopram significantly antagonizes bax mRNA and potentiatesbcl-2 mRNA protein expression and inhibits apoptosis of rat prefrontal cortical neurons caused by chronic stress, which might be one possible mechanism of Citalopram for prevention and treatment of psychosis caused by chronic stress.

Animals ; Apoptosis ; Citalopram ; pharmacology ; Male ; Neurons ; drug effects ; metabolism ; Prefrontal Cortex ; cytology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Stress, Physiological ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo study the distribution laws of the Chinese medicine syndrome patterns and its correlated symptoms in patients with polycystic ovarian syndrome (PCOS), and the possible correlation between Chinese medicine syndrome patterns and PCOS associated parameters, thus to provide a guidance for selecting proper indices in curative effectiveness assessment.

METHODSUsing clinical epidemiological methods and mathematical statistics, the Chinese medicine syndrome patterns were studied in 228 PCOS patients. The distribution features of Chinese medicine syndrome patterns were summarized.

RESULTSShen-deficiency blood-stasis syndrome was the most frequently seen in PCOS patients, followed by Pi-deficiency phlegm-dampness syndrome, Pi-Shen yang-deficiency syndrome, and Shen-yin deficiency syndrome. Positive correlation existed between serum levels of follicle stimulating hormone (FSH) and Pi-Shen yang-deficiency syndrome. Positive correlation existed between fasting blood sugar (FBS), waist to hip ratio (WHR), body mass index (BMI), fasting insulin (FIN), and Hirsutism score and Pi-deficiency phlegm-dampness syndrome. Positive correlation existed between serum levels of prolactin (PRL), estradiol (E2 ) and qi stagnancy and blood stasis syndrome. Positive correlation existed between serum progesterone (PRG) level, FSH, FIN, BMI, acne score and Gan stagnancy and blood heat syndrome. Positive correlation existed between luteinizing hormone (LH) and Shen-deficiency blood-stasis syndrome. Besides, LH/FSH >3 was possibly more frequently seen in Pi-deficiency phlegm-dampness syndrome and Pi-Shen yang-deficiency syndrome. Family heritability could be seen in each syndrome patterns. Among them, female heritability was more often seen in Shen-deficiency blood-stasis syndrome, while male heritability was more often seen in Shen-yin deficiency syndrome.

CONCLUSIONSShen-deficiency blood-stasis syndrome, Pi-deficiency phlegm-dampness syndrome, Pi-Shen yang-deficiency syndrome, and Shen-yin deficiency syndrome were most frequently seen in PCOS patients. The sex hormones (including 6 items), FBS, FIN, WHR and BMI, etc. were correlated with each Chinese medicine syndrome pattern to various extents, which could be taken as reference in Chinese medicine syndrome differentiation.

Adult ; Body Mass Index ; Female ; Humans ; Medicine, Chinese Traditional ; Polycystic Ovary Syndrome ; diagnosis

OBJECTIVETo investigate the risk factors for malignant transformation of oral leukoplakia.

METHODSA total of 409 cases with oral leukoplakia was retrospectively analyzed. Single factor test was first performed to examine the associations between oral leukoplakia's histopathological classification and each of risk factors including sex, age, systemic diseases, course of disease, clinical classification, site, size, numbers of lesion, alcohol and tobacco consumption, and symptom. Then the association of these selected factors with oral leukoplakia's histopathological classification was evaluated using multiple logistic regression analysis.

RESULTSFifty-two cases of all 409 patients with oral leukoplakia (including 9 severe dysplasia) developed oral cancer. The ratio of malignant transformation was 12.7%. Sex, age, clinical type, site and symptom were chosen as risk factors incorporated into the multiple logistic regression models. The risk of mild-moderate dysplasia in female was 2.40 times as high as that in male. The risk of mild-moderate dysplasia of speckled leukoplakia was 2.81 times as high as that of homogeneous leukoplakia. The risk of mild-moderate dysplasia of dangerous site was 1. 98 times as high as that non-dangerous site. The risk of mild-moderate dysplasia with symptom was 1.84 times as high as that without symptom. The risk of severe dysplasia and oral cancer in female was 3.11 times as high as that in male. The risk of severe dysplasia and oral cancer of speckled (4.50 times), ulcerative (5.63 times), verrucous leukoplakia (4.09 times) were much higher than that of homogeneous leukoplakia. The risk of severe dysplasia and oral cancer in dangerous site was 2.79 times as high as in non-dangerous site. The risk of severe dysplasia and oral cancer in leukoplakia with symptom was 4.38 times as high as without symptom.

CONCLUSIONSThe malignant transformation of oral leukoplakia is correlated to sex, clinical type, site and symptom.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Leukoplakia, Oral ; etiology ; Male ; Middle Aged ; Mouth Neoplasms ; etiology ; Retrospective Studies ; Risk Factors ; Young Adult

Background Following the accelerated speed of population aging in China,the incidence of cataract is rising gradually.Researches indicated that senescenee marker protein-30 ( SMP-30 ) is closely associated with the occurring and developing of cataract. Objective This study was to investigate the expression of SMP-30 in human lens epithelial cells(LECs) of age-related cataract and study the relationship between SMP-30 and apoptosis.Methods Capsulotomy was performed on 80 eyes of 59 patients with simple cortex age-related cataract and 70 eyes of 53 age-matched patients with nucleus age-related cataract.The anterior capsular specimens were obtained by circularly capsulorhexis during the operation.Expressions of the SMP-30 protein and mRNA in the LECs of two types of cataract were detected using immunochemistry and real-time PCR respectively.Apoptosis of the LECs was assayed by TUNEL.The differences of expression of SMP-30 and apoptosis were compared between the two types of cataract.Results Immunochemistry showed that SMP-30 was expressed in cytoplasm of LECs.The expression intensity of SMP-30 was higher in the center zone compared with periphery zone.The apoptosis rate of LECs was significantly higher in the center of the anterior capsule than the periphery in both two types of cataract ( nucleus cataract:19.34％±0.11％ vs 8.32 ％ ± 0.57 ％,P =0.025 ; cortex cataract:42.07 ％ ± 0.86 ％ vs 13.55 ％ ± 0.64 ％,P =0.010 ).The expression amount of SMP-30 mRNA was lower at the periphery than the center of lens in both two types of cataract (nucleus cataract:45.21±2.79 vs 76.42±11.21,P=0.042 ;cortex cataract:108.32±4.32 vs 206.34±15.67,P=0.037 ),and that of nucleus cataract was significantly lower than cortex cataract (60.02±9.08 vs 157.33 ± 13.01,P =0.034),and the apoptosis rate of LECs was declined in the nucleus cataract group compared with the cortex cataract group ( 14.05％ ±0.22％ vs 27.70％ ±0.81％,P =0.007 ). Conclusions LECs apoptosis exists in age-related cataract.SMP-30 probably plays an important role in the formation of cataract.

OBJECTIVETo investigate the correlation between the number and activity of circulating endothelial progenitor cells (EPCs), insulin resistance and severity of coronary lesions in patients with coronary artery disease (CAD).

METHODSPatients with coronary angiography evidenced CAD were divided in insulin resistance group (IR, n = 25) and insulin sensitive group (IS, n = 44) according to insulin level, 25 health volunteers served as control. Circulating EPCs were marked as KDR/CD133+ cells via fluorescence-activated cell sorter analysis. EPCs were also isolated from peripheral blood and cultured in vitro for 7 days, identified by DiI-acLDL uptake and lectin staining methods. EPCs migration activities were determined by modified Boyden chamber assay, EPCs proliferation activities were determined by MTT assay.

RESULTCirculating EPCs number was significantly lower in IR group compared with IS group [(0.34 +/- 0.08) per thousand vs. (0.47 +/- 0.09) per thousand, P < 0.01] and control group (P < 0.05). Both insulin resistance index (r = -0.291, P = 0.01)and Gensini score (r = -0.3984, P = 0.006)were negatively correlated with number of circulating EPCs. Proliferation and migration capacities of EPCs were also significantly lower in IR group compared to those in IS group (all P < 0.05) and control group (all P < 0.05).

CONCLUSIONSInsulin resistance/hyperinsulinemia could aggravate severity of coronary artery lesions via reducing the number and activities of circulating EPCs in patients with CAD.

Aged ; Blood Cell Count ; Cell Adhesion ; Cell Movement ; Cell Proliferation ; Cells, Cultured ; Coronary Angiography ; Coronary Artery Disease ; blood ; pathology ; Endothelial Cells ; cytology ; Female ; Humans ; Insulin Resistance ; Male ; Middle Aged ; Stem Cells ; cytology

The aim of this study was to explore the effects of cholecystokinin octapeptide (CCK-8) on cardiac function and the receptor mechanism in anesthetized rats. The mean arterial pressure (MAP), heart rate (HR), the left ventricle systolic pressure (LVP) and the maximal/minimum rate of LVP (+/-LV dp/dt(max)) were measured. The results obtained are as follows. (1) Low dose of CCK-8 (0. 4 microgram/kg i.v.) caused tachycardia and slight increase in MAP, LVP and LV dp/dt(max) (P<0.01), while medium dose (4.0 microgram/kg i.v.) and high dose of CCK-8 (40 microgram/kg i.v.) elicited a bradycardia and marked increase in MAP, LVP and LV dp/dtmax (P<0.01). (2) Proglumide (1.0 mg/kg i.v.), a CCK-receptor (CCK-R) antagonist, significantly inhibited the pressor effects of CCK-8, whilst it reversed the bradycardic responses (P<0.01). (3) Using reverse transcription polymerase chain reaction (RT-PCR), CCK-A receptor (CCK-AR) and CCK-B receptor (CCK-BR) mRNA were expressed in myocardium of rats. The above results indicate that CCK-8 may enhance cardiac function in a dose-dependent manner and elicit a change in HR, which is likely induced by the activation of CCK-R on myocardium.
Animals ; Dose-Response Relationship, Drug ; Heart Rate ; drug effects ; Male ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Cholecystokinin ; drug effects ; Sincalide ; administration & dosage ; pharmacology ; Ventricular Function, Left ; drug effects ; Ventricular Pressure ; drug effects

To study the effect of cholecystokinin-octapeptide (CCK-8) on systemic hypotension and cytokine production in serum and lung of endotoxic shock (ES) rats induced by lipopolysaccharide (LPS) and investigate its signal transduction mechanism of p38 mitogen-activated protein kinase (MAPK), the changes in mean arterial pressure (MAP) were observed by using a polygraph in four groups of SD rats: group of LPS (8 mg/kg i.v.) induced ES, group of CCK-8 (40 microg/kg i.v.) pretreatment 10 min before LPS (8 mg/kg) administration, group of CCK-8 (40 microg/kg i.v.) only, and normal saline (control) group; the contents of proinflammatory cytokines (TNF-alpha, IL-1 beta and IL-6) in the lung and serum were assayed using ELISA kits; and p38 MAPK was detected by Western blot. The results showed that CCK-8 alleviated LPS-induced decrease in MAP of rats; compared with the control, LPS elevated the levels of TNF-alpha, IL-1 beta and IL-6 in serum and lung significantly, while CCK-8 significantly inhibited the LPS-induced increases in TNF-alpha, IL-1 beta and IL-6 in serum and lung. The activation of p38 MAPK in the lung of ES rats was enhanced by CCK-8 pretreatment. These results suggest that CCK-8 can alleviate the LPS-induced decrease in MAP of ES rats and exert an inhibitory effect on the overproduction of proinflammatory cytokines, and that p38 MAPK may be involved in its signal transduction mechanisms.
Animals ; Cytokines ; biosynthesis ; Lung ; metabolism ; Male ; Mitogen-Activated Protein Kinases ; biosynthesis ; physiology ; Rats ; Rats, Sprague-Dawley ; Shock, Septic ; drug therapy ; metabolism ; Sincalide ; pharmacology ; p38 Mitogen-Activated Protein Kinases

OBJECTIVETo investigate the development and maturation competence of oocytes retrieved from cryopreserved and transplanted human fetal ovarian tissue by techniques of tissue culture, inducing ovary, oocyte retrieval, and in vitro maturation (IVM).

METHODSFetal ovaries of 20 weeks were frozen-thawed and cultured for 6 days in vitro, then xenografted into kidney capsules of immunodeficient mice. All mice were stimulated with follicle stimulating hormone every second day for 23 weeks, starting 1 week after grafting. Then oocytes were retrieved from antral follicles 13 hours after human chorionic gonadotrophin injection. IVM was performed to evaluate the maturation competence of the oocytes from ovarian grafts. Human fetal ovarian tissues were examined with histological and proliferating cell nuclear antigen (PCNA) evaluation.

RESULTSThere was no difference between fresh ovarian tissues and frozen-thawed ovarian tissues in the percentage of follicles at different growth stages (P > 0.05). The proportion of the primary follicles and preantral follicles in the cultured ovarian tissues was significantly larger than that of fresh ovarian tissues and frozen-thawed ovarian tissues (P < 0.05). The proportion of the primary follicles, preantral follicles, and antral follicles in the transplanted ovarian tissues was significantly higher than that of cultured ovarian tissues, fresh ovarian tissues, and frozen-thawed ovarian tissues (P < 0.05). No significant signals of PCNA in the primordial follicles in all ovarian tissues were observed. PCNA immunoreactivity first appeared in primary follicles. However, the obviously positive signals of PCNA were seen in the oocytes and/or the granular cells of cultured ovarian tissues and transplanted ovarian tissues. Oocytes from antral follicles were collected and matured in vitro, and 21.43% of the oocytes reached to MII within 48 hours IVM.

CONCLUSIONSHuman ovarian follicles can survive and develop well after cryopreservation, tissue culture, and xenotransplantation. Furthermore, oocytes recovered from grafts have normal maturation competence.

Animals ; Cryopreservation ; Female ; Humans ; Mice ; Mice, Inbred BALB C ; Oocytes ; cytology ; Oogenesis ; Ovarian Follicle ; cytology ; growth & development ; transplantation ; Pregnancy ; Transplantation, Heterologous

OBJECTIVETo explore the role of bone morphogenetic protein-7 (BMP-7) in strontium ranelate (Sr)-induced osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs).

METHODSBMSCs were isolated from 4-week-old rats and cultured in vitro. The third or fourth passages of BMSCs were examined using alkaline phosphatase kit for changes in ALP activity in response to treatment with different concentrations of Sr. Calcium nodules in the induced cells were detected using alizarin red staining, and the cellular BMP-7 expression was detected by Western blotting.

RESULTSWithin the concentration range of 0.1-3.0 mmol/L, Sr dose-dependently increased ALP activity in rat BMSCs. ALP activity reached the highest level after treatment with 3 mmol/L Sr, which also significantly promoted the formation of calcium nodules. Within the range of 0.1-3.0 mmol/L, Sr dose-dependently enhanced the expression of BMP-7, and its peak expression occurred following 3 mmol/L Sr treatment. Noggin (100 ng/ml), an inhibitor of BMP-7, obviously suppressed Sr-induced over-expression of BMP-7 and reduced ALP activity and calcium nodule formation in the BMSCs.

CONCLUSIONSr promotes osteogenic differentiation of rat BMSCs by increasing the expression of BMP-7.

Animals ; Bone Density Conservation Agents ; pharmacology ; Bone Marrow Cells ; cytology ; Bone Morphogenetic Protein 7 ; genetics ; metabolism ; Cell Differentiation ; drug effects ; Cells, Cultured ; Female ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Organometallic Compounds ; pharmacology ; Osteoblasts ; cytology ; Osteogenesis ; drug effects ; Rats ; Thiophenes ; pharmacology