OBJECTIVETo observe the effect of Pinggan Qianyang Recipe (PQR) on inhibiting angiotensin II (Ang II) induced proliferation and migration of vascular smooth muscle cells (VSMCs) and changes of DNA methylation.

METHODSVSMCs were cultured using tissue explant method, and PQR containing serum was prepared. Primarily cultured VSMCs were divided into four groups, the normal group, the model group, the folate group (folic acid intervention) , and the PQR group. The proliferation and migration of VSMCs was duplicated by Ang II. After 24-h Ang II induced culture, 40 microg/mL folic acid was added to the folate group for 48 h, while 5% PQR containing serum was added to the PQR group for 48 h. The cell growth curve of VSMCs was drawn by using Cell Counting Kit (CCK-8). The proliferative activity of VSMC was determined by MTT assay. The migration of VSMCs was measured by Millicell chamber. The general level of cytosine methylation in cell nucleus was detected via 5-mC antibodies immunofluorescence, and mRNA expression levels of DNA methyltransferase 1 (DNMT1) were measured by Real-time q-polymerase chain reaction (q-PCR).

RESULTSVSMCs were promoted by Ang II at 10(-6) mol/L for 24 h. Compared with the normal group, the proliferative activity and migration quantity of VSMCs obviously increased, and DNA methylation level obviously decreased (P < 0.05, P < 0.01). Compared with the model group, the cell growth, proliferative activity and migration quantity of VSMCs obviously decreased and the general DNA methylation level increased in the folate group and the PQR group (P < 0.05, P < 0.01). Compared with the normal group, the mRNA expression of DNMT1 decreased in the model group (P < 0.01). Compared with the model group, mRNA expression of DNMT1 in Ang II induced VSMCs was obviously enhanced in the folate group and the PQR group (P < 0.01).

CONCLUSIONSPQR could inhibit Ang II induced proliferation and migration of VSMCs, and cause high genomic DNA methylation level. Changes of DNA methylation might be associated with DNMT1 expression.

Angiotensin II ; pharmacology ; Cell Movement ; Cell Proliferation ; Cells, Cultured ; DNA (Cytosine-5-)-Methyltransferase 1 ; DNA (Cytosine-5-)-Methyltransferases ; metabolism ; DNA Methylation ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; drug effects

OBJECTIVETo study the impact of the water extract from Codonopsis thalictrifolia Wall (CTW) on the reproductive

METHODSWe divided 32 male SD infant rats into four groups of equal number to be treated intragastrical-system of male infant rats. ly with distilled water (control) and CTW at 10 g/kg (low dose) , 20 g/kg (medium dose), and 40 g/kg (high dose), respectively, twice a day for 2 weeks. Then we killed the rats, measured the levels of testosterone (T), luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in the serum, obtained the testis weight, body weight, testis visceral coefficient and sperm concentration, and detected sperm viability, sperm motility and the level of cyclic adenosine monophosphate (cAMP) in the Leydig cells, followed by

RESULTSCompared with the control group, the low-dose, me-analysis of differences among different groups using the SPSS software. Medium-dose and high-dose CTW groups showed significant decreases in the serum T level ([3.09 +/-0.42] vs [1.22 +/-0. 32] , [1.06 +/- 0.29] and [0.57 +/-0.18] nmol/L, P<0.01), testis weight ([1.40 +/-0.16] vs [0.96 +/-0.09], [0.92 +/-0.11] and [0.91 +/- 0.08] g, P <0.01), and sperm concentration ([1.03 +/-0.16] vs [0.19 +/-0.07], [0.17 +/-0.08] and [0.16 +/-0.07] x 10(6)/ml, P <0.01), but a dramatic elevation in the testis visceral coefficient ([42.22 +/- 3.02] vs [51.39 +/- 3.09], [52.28 +/- 4.86] and [54.13 +/-6.06] mg/10 g, P <0.01); the medium- and high-dose CTW groups exhibited remarkable increases in the levels of serum LH ([13.62+/-0.89] vs [14.69 +/-0.12] and [14.93 +/-0.28] ng/L, P<0.01) and FSH ([4.32 +/-0.18] vs [4.77 +/-0.23] and [4.89 +/-0. 38] IU/L, P <0.05); all the three CTW groups showed markedly inhibited serum T secretion ([1.85 +/- 0.18] vs [1.42 +/-0.15], [1.12+/-0.18] and [0.88 +/-0.21] nmol/L, P<0.01) and intracellular cAMP ([5.51 +/-0.12] vs [4.39+/-0.06], [4.28 +/-0.07] and [4.11 +/- 0.10] nmol/L, P <0.01) in the Leydig cells.

CONCLUSIONThe water extract from CTW may reduce the synthesis of testosterone in the serum of male infant rats through the PKA pathway and consequently inhibit their testicular development and sperm production and affect the development of their reproductive system.

Animals ; Codonopsis ; chemistry ; Cyclic AMP ; metabolism ; Leydig Cells ; metabolism ; Male ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Testosterone ; blood ; Urogenital System ; drug effects

Tumor immune checkpoint therapy is a clinical treatment strategy developed based on the new principle of the inhibition of negative immune regulation. In this article, the tumor immune checkpoint therapy and the drug delivery strategies were reviewed, mainly including immunity and tumor therapy, tumor immune checkpoint therapy and its mechanism of action, clinical application of tumor immune checkpoint therapy and therapeutic drugs, immune resistance of programmed cell death protein 1 (PD1)/programmed cell death ligand 1 (PDL1) treatment and countermeasures, drug delivery strategies for tumor immune checkpoint therapeutic agents, etc. As a revolutionary new immunotherapy strategy, tumor immune checkpoint therapy has shown obvious superior therapeutic efficacy in a variety types of tumor. However, tumor immune checkpoint therapy is also faced with a big challenge, namely, immunotherapy resistance. With the discovery of new mechanism, the continuous development of new therapeutic drugs and delivery strategies, tumor immune checkpoint therapy is expected to further improve the clinical efficacy of tumor.

The study was aimed to investigate the genetic background and proliferation characteristics of multiple myeloma (MM). Myeloma cells were isolated from bone marrow of 19 MM patients by direct immunomagnetic cell sorting and the DNA content and cell cycle analysis were carried out by flow cytometry. The results showed that in 4 patients the myeloma cells were found to be hyperdiploid and in 15 patients those were found to be diploid respectively by DNA content analysis; the proportion of plasm cells from normal controls in S + G(2)/M phase was (1.15 +/- 0.60)%, and that of myeloma cells from MM patients was (10.06 +/- 12.60)% which was significantly higher than that in the former (p = 0.001). The incidence of hyperdiploid in newly diagnosed patients was 11.76%, and that of treated patients was 100.00% which was significantly higher than that in the former (p = 0.035); the proportion of myeloma cells from newly diagnosed patients in S + G(2)/M phase was (7.12 +/- 4.98)%, and that of treated patients was (35.10 +/- 32.56)% which was also significantly higher than that in the former (p = 0.001). It is concluded that the variety of myeloma cells in DNA content and cell cycle suggests the complicated genetic background and abnormal proliferation of MM, which relate with the course of disease to some extent.
Adult ; Aged ; Aged, 80 and over ; Bone Marrow Cells ; metabolism ; pathology ; Cell Cycle ; genetics ; Cell Proliferation ; DNA ; analysis ; genetics ; Diploidy ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; pathology

OBJECTIVETo investigate the clinical significance of the deletion of the long arm of chromosome 13 [del (13 q) ] and translocation of immunoglobulin heavy chain gene [t (14 q) I in multiple myeloma (MM) patients.

METHODSMyeloma cells were isolated from hone marrow by direct immunomagnetic cell sorting and interphase fluorescence in situ hybridization (FISH) was performed in 24 MM patients to detect del (l3q) and t (l4q).

RESULTSThe positive rates of del (l3q) and t (l4q) were 45.83% and 37.50% respectively. Five patients (20.83%) had both two abnormalities and 15 patients (62.50%) had at least one abnormality. Univariate analysis showed that the positive rates of del (l3q) were 35.71% and 66.67% in responders and non-responders (P = 0.214) and the positive rates of t (l4q) were 21.43% and 66. 67% in responders and non-responders (P = 0.077). Multivariate analysis showed that del (13q) (OR = 5.761, 95% CI 0.500-66.391, P = 0.160), t (14q) (OR = 6.576, 95% CI 0.580-74.614, P = 0.129), and corrected serum calcium level (OR = 8.080, 95% CI 0.738-88.427, P = 0.087) were relatively independent negative factors for response to therapy, with the corrected serum calcium level being the strongest reversely-correlated factor.

CONCLUSIONSInterphase FISH is a sensitive method to investigate the cytogenetics of MM. Del (13q), t (14q), and corrected serum calcium level can be used to predict treatment response and prognosis.

Adult ; Aged ; Aged, 80 and over ; Chromosome Deletion ; Chromosomes, Human, Pair 13 ; genetics ; Chromosomes, Human, Pair 14 ; genetics ; Female ; Humans ; Immunoglobulin Heavy Chains ; genetics ; In Situ Hybridization, Fluorescence ; Interphase ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; Translocation, Genetic

OBJECTIVETo compare the effects of amlodipine, benidipine and nifedipine on myocardial hypertrophy and evaluate the underlying mechanism.

METHODSMyocardial hypertrophy model was created by transverse aortic constriction (TAC) in C57 BL/6 mice, and plasma catecholamine concentrations were measured 7 days after surgery to confirm the sympathetic activation. The 3 drugs were administered in TAC mice for 7 days and cardiac hypertrophy was evaluated according to the heart-to-body weight ratio (HW/BW). Effects of those drugs on the protein synthesis stimulated by phenylephrine in cultured neonatal cardiac myocytes were also examined.

RESULTSHW/BW and plasma concentrations of catecholamine were significantly increased in TAC mice one week after surgery in comparison with to sham-operated mice. One week after TAC, the HW/BW ratio was significantly lower in the amolodipine but not nifedipine-treated group than in the TAC group. Administration of nifedipine via minipump infusion for one week did not decrease HW/BW ratio. Treatment with amlodpine or benidipine, but not nifedipine, decreased the neonatal rat myocyte protein synthesis induced by phenylephrine stimulation.

CONCLUSIONAntihypertrophic effect of DHEs on myocardium is dependent on their potential of blocking N-type calcium channel, and the underlying mechanism involves the sympathetic inhibition.

Amlodipine ; pharmacology ; therapeutic use ; Animals ; Calcium Channel Blockers ; pharmacology ; therapeutic use ; Calcium Channels, N-Type ; drug effects ; Cardiomegaly ; drug therapy ; etiology ; Dihydropyridines ; pharmacology ; therapeutic use ; Disease Models, Animal ; Male ; Mice ; Mice, Inbred C57BL ; Nifedipine ; pharmacology ; therapeutic use

Objective To study the effect of metformin on glucolipid metabolic disorders and liver lipid deposition caused by clozapine in rats.Methods From 1 d to 4 d,Clozapine 5 mg·kg -1 ·d -1 was gavaged,and the dose increased to 25 mg·kg -1 ·d -1 from the 5th day.Metformin 100 mg·kg -1 ·d -1 or 400 mg·kg -1 ·d -1 or simvastatin 1 mg· kg -1 ·d -1 was gavaged from the 15th day.The total period of dosing was 8 weeks.Body mass,fasting blood sugar (FBS) and postprandial 2 hours blood glucose (2hPBG)were measured at baseline,3 d,1 week,2 weeks,4 weeks,6 weeks and 8 weeks.At the end of the 8th week,serum cholesterol (TC),triglyceride (TG),low density lipoprotein (LDL -C), high density lipoprotein (HDL -C),fructosamine (FA)and insulin (IRS)were measured and liver HE staining was done.Results There were no significant differences of the measured indexes between control group and metformin group at the all test points.By the end of the 6th and 8th week,compared with control group,the body mass,FBS,2hPBG,IRS, FA,TC,TG and LDL -C were significantly increased in clozapine group (P <0.05 ),while HDL -C decreased in clozapine group (P <0.05).Compared with clozapine group,body mass,FBS,2hPBG,IRS,FA,TC,TG and LDL -C were significantly decreased by metformin or simvastatin administration (P <0.05),while HDL -C increased(P <0.05).Rat liver cells in clozapine group were not neat around the small blood vessels;there were more white fat cells and hepatocellular lipid calm far away from the blood vessels.However,in other groups,there were moderate white fat cells, and there were not much hepatocellular lipid calm far away from the blood vessels.Conclusion Metformin could effectively prevent and treat weight gain,glucolipid metabolic disorder and liver lipid deposition caused by clozapine.

OBJECTIVETo probe the regulative effect of electroacupuncture (EA) at "Fenglong" (ST 40) on blood lipids in hyperlipidemia (HLP) rats and the mechanism.

METHODSEighty Wistar rats were randomly divided into a normal group (fed with basal forage), a model group (fed with high fat forage), an EA group (fed with high fat forage + EA treatment), a western medicine group (fed with high fat forage + Pravastatin sodium). Contents of serum total cholesterol (TC), triacylglycerol (TG), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), endothelin (ET), nitric oxide (NO) and calcitonin gene-related peptide (CGRP) were determined before and after treatment.

RESULTSCompared with the normal group, the body weight, levels of TC, TG, LDL-C and ET were significantly elevated (P < 0.05, P < 0.01) and the levels of HDL-C, NO and CGRP were significantly decreased (P < 0.05) in the model group; compared with the model group, the body weight, levels of TC, TG and LDL-C were significantly decreased (P < 0.01) and the levels of NO and CGRP were significantly increased in the western medicine group and the EA group (P < 0.01, P < 0.05); compared with the EA group, HDL-C level significantly increased in the western medicine group (P < 0.01), and ET level decreased in the EA group and the western medicine group with no significant difference between the two groups (P > 0.05).

CONCLUSIONBoth EA and Pravastatin sodium have better benign regulative effects on TC, TG, LDL-C, NO and CGRP and can decrease ET level to a certain extent in the rat of hyperlipidemia.

Acupuncture Points ; Animals ; Calcitonin Gene-Related Peptide ; blood ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Electroacupuncture ; Endothelins ; blood ; Hyperlipidemias ; blood ; therapy ; Male ; Nitric Oxide ; blood ; Rats ; Rats, Wistar

OBJECTIVESTo investigate the role of focal adhesion kinase (FAK) in the pathogenesis of cardiac hypertrophy induced by hypertension.

METHODSUsing immunofluorescent labeling, confocal microscopy and Western blotting, the expression and subcellular localization of FAK in the cardiac myocytes of left ventricle were determined in 2, 6, 12, and 18 month-old rats with spontaneously hypertensive heart failure (SHHF) along with age-matched control Wistar-Kyoto (WKY) rats.

RESULTSThere was no significant difference of FAK expression between 2 month-old SHHF and WKY rats (50.5+/-6.9 vs. 49.8+/-5.0, n=6, P>0.05). In contrast with the control groups, the expression of FAK significantly increased in 6, 12 and 18 month-old SHHF rats (130.6+/-3.0 vs. 47.3+/-1.3, 144.7+/-5.4 vs. 46.4+/-3.1, 141.4+/-9.8 vs. 48.5+/-2.2, each groups n=6, P<0.05) with FAK protein primarily cumulated in the intercalated disks and nuclei.

CONCLUSIONSFAK may play a role in the cell signaling transduction leading to cardiac hypertrophy, presumably through regulations of hypertrophic gene transcription and RNA processing.

Animals ; Focal Adhesion Kinase 1 ; metabolism ; Heart Ventricles ; pathology ; Hypertension ; complications ; Hypertrophy, Left Ventricular ; enzymology ; etiology ; Male ; Microscopy, Confocal ; Myocytes, Cardiac ; metabolism ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Signal Transduction

OBJECTIVE: To study the expression and effect of Pim1 in primary cortical neurons after hypoxic-ischemic injury. METHODS: Cortical neurons were isolated from 1-day-old C57BL/6 mice and cultured in neurobasal medium. On the 8th day of neuron culture, cells were subjected to oxygen-glucose deprivation/reoxygen (OGD/R) treatment to mimic in vivo hypoxic injury of neurons. Briefly, medium were changed to DMEM medium, and cells were cultured in 1% O for 3 hours and then changed back to normal medium and conditions. Cells were collected at 0 hour, 6 hours, 12 hours and 24 hours after OGD/R. Primary neurons were transfected with Pim1 overexpression plasmid or mock plasmid, and then were exposed to normal conditions or OGD/R treatment. They were named as Pim1 group, control group, OGD/R group and OGD/R+Pim1 group respectively. Real-time PCR was used to detect Pim1 mRNA expression. Western blot was used to detect the protein expression of Pim1 and apoptotic related protein cleaved caspase 3 (CC3). TUNEL staining was used to detect cell apoptosis. RESULTS: Real-time PCR and Western blot results showed that Pim1 mRNA and protein were significantly decreased in neurons after OGD/R. They began to decrease at 0 hour after OGD/R, reached to the lowest at 12 hours after OGD/R, and remained at a lower level at 24 hours after OGD/R (P<0.01). Overexpression of Pim1 significantly upregulated the protein level of Pim1. Under OGD/R conditions, the CC3 expression and the apoptosis rate in cells of the Pim1 group were significantly lower than in un-transfected cells (P<0.01). CONCLUSIONS Hypoxic-ischemic injury may decrease Pim1 expression in neurons. Overexpressed Pim1 may inhibit apoptosis induced by OGD/R.
Animals ; Glucose ; Mice ; Mice, Inbred C57BL ; Neurons ; Oxygen ; Proto-Oncogene Proteins c-pim-1 ; Rats, Sprague-Dawley