Animals ; Humans ; Models, Animal ; Myocytes, Cardiac ; cytology ; Stem Cells ; cytology ; Tissue Engineering

Objective To investigate the change of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the skin of diabetic rats, and to explore the potential role of MMP-9/TIMP-1. Methods Diabetic rats were induced with streptozotocin (STZ). Then all rats were maintained for 6 weeks. Routine pathological examination and immnnohistochemistry were made to reveal the histological and cytological appearances. RT-PCR and Western blotting were used to detect the expression of mRNA and protein of MMP-9 and TIMP-1 in the skin. Results Six weeks after STZ treatment, examination of HE-stained skin sections from normal and diabetic animals revealed that the epidermis and dermis layers were thinner in diabetic rats than those in control rats. The skin of diabetic rats showed features of atrophy such as disorganization of connective tissue fiber bundles and enlarged space between collagen fiber bundles. In contrast, thick bundles of connective tissue were observed in the dermis of normal rat skin. In normal skin, cells had a bipolar, spindle-shaped appearance in the thick collagen bundles, while in the skin of diabetic animals the interstitial cells had a rounded, shrunken and crenated appearance. The relative values of expression of MMP-9 in diabetic group were higher than those in normal group with significant difference, however, the relative values of expression of TIMP-I in diabetic group were lower than those in control group. Conclusion The changes in cutaneous histology and cytology appear earlier than skin wound. These "underlying disorders" may be associated with the imbalance of MMP-9/TIMP-1.

AIM: To observe the effects of six common traditional Chinese herbs of activating blood, paeoniae rubra radix, salviae miltiorrhizae radix, ligustici, rhizome, notoginseng radix, pruni persicae semen and wine staemed radix et rhizome, on atherosclerotic plaque structure and stabilization in ApoE gene-deficient mice. METHODS: Four sections of the aortic root were choosen and stained with hematoxylin and masson. All sections were measured with Image-ProDR○ Plus Version 4.5.1 (IPP) software. RESULTS: Compared with the model group, plaque area corrected by cross-sectional vessel wall area reduced significantly in salviae miltirrhizae radix treatment group, lipid core area reduced in paeoniae rubra radix group, pruni persicae semen and wine steamed radix et rhizome treatment group, minimum thickness of fibrous cap became thicker significantly in salviae miltiorrhizae radix, ligustici, rhizome, pruni persicae semen and wine steamed radix et rhizome treatment group. CONCLUSION: These Chinese herbs may stabilize the atherosclerotic plaques in ApoE gene-deficient mice by interfering their structure, but their effects do not parallel with their activating blood efficacy in traditional Chinese medicine.

Background Channelrhodopsin-2 (ChR2)is a cation channel isolated from the eyespot of Chlamydomonas algae and has been used to control neuron activity.The light stimulation is a more precise fashion whether space or time than that of electrical,magnetic and ultrasound stimulation. Objective This study was to construct a replication deficient recombinant adenovirus cxpression vector of ChR2 and to determine its function.Methods Human embryo kidney 293 (HEK293) cell line was cultured and passaged in DF12 medium containing 10％ fetal bovine serum(FBS).The ChR2 gene was cloned at the downstream of cytomegalovirus(CMV)promoter of the adenoviral shuttle plasmid pSB291 in sense direction,and the resultant recombinant plasmid pSB291-hChR2- GFP was transfected into HEK293 cell together with plasmid pBHG lox ( deltaE1,3 ) containing adenoviral genome,then small amounts replication deficient recombinant adenovirus expression vector of ChR2 (Ad-ChR2) was obtained.Through amplification gradient centrifugation and dialysis,pure Ad-ChR2 was obtained.Visual cortex cells derived from 4 1-day-old clean Long Evans rats were primary cultured with serum-free culture media and infected by AdChR2.When expressing green fluorescencc,those cells received the stimulated of blue light with 460 nm.Patch clamp technique was applied to record an action potential. Results After purification and concentration,the titer of AdhCHR2 reached 7.9×1010 PFU/ml.Twenty-four hours after transfect of Ad-ChR2,HEK293 cell membrane showed the green fluorescence for the recombinant plasmid with green fluorescence protein under the inversed fluorescence microscope.The HEK293 cells change their shape from flat to round 13 days after transfected.The primary cultured visual cortex cells exhibited the green fluorescence 3-5 days after infected by Ad-ChR2.The action potentials evoked by blue light stimulation were recorded with patch clamp on those cells expressing green fluorescence. Conclusions Ad-ChR2 expressing vector is constructed successfully in this study.It is verified that Ad-ChR2 expressing vector can infect visual cortex cells with visual function.This result is very important for visual plasticity study.

OBJECTIVETo investigate plasma endothelin-1 (ET-1) level in patients with prostate cancers and its clinical significance.

METHODSPlasma ET-1 level was measured by radioimmunoassay in 31 patients of prostate cancer (23 with non-HRPC, 8 with HRPC) and 26 patients of BPH.

RESULTSCompared with each other of the ET-1 level, there were no significant difference among the BPH group,non-HRPC group and HRPC group. No significant difference was found either between bone metastasis (BM) and non- BM, between high and middling differentiation prostate cancer group, as well as in different PSA level groups (P >0.05). But the ET-1 level in low differentiation prostate cancer was notably lower than those of the high and middle respectively (P < 0.05).

CONCLUSIONTo detect plasma endothelin-1 (ET-1) level is not a useful method to evaluate the development and the prognosis of prostate cancer.

Aged ; Aged, 80 and over ; Endothelin-1 ; blood ; Humans ; Male ; Prognosis ; Prostatic Hyperplasia ; blood ; Prostatic Neoplasms ; blood ; Radioimmunoassay

Objective To analyze the data of road traffic accidents, deaths and injuries in 31 provinces and municipalities in China from 2010 to 2017, and to predict that the number of deaths and injuries caused by road accidents set in the sustainable development goals (SDGs) will be halved by 2020 compared with the target set in 2015. Methods Describing and analyzing the current situation of road traffic accidents in 31 provinces and municipalities in China, and using the trend extrapolation method to predict whether the SDGs target can be achieved by 2020. Results The numbers of traffic accidents, deaths and injuries in Guangdong, Jiangsu, Zhejiang and Shandong Provinces showed a downward trend from 2010 to 2017, but still ranked the front in China. In 2017, Beijing had the highest death rate followed by Guizhou as the second, and Guizhou had the highest injury rate followed by Tianjin as the second. The projected results showed that the numbers of deaths and injuries caused by road traffic accidents in China would be increased by 17% and 1.3% respectively in 2020 indicating that SDGs target could not be met. Among the 31 provinces/municipalities, it was predicted that the numbers of deaths and injuries in Hunan would be reduced to 50.7% and 65.3% in 2015, respectively by 2020, and the target could be achieved; the number of injuries in Shanghai was decreased by 83.3%, but the deaths was only decreased by 34.5%, and there was still a gap with the target; the rest 29 provinces/municipalities could not meet SDGs target. It was expected that the deaths in 11 provinces and municipalities would increase by 2020, with Beijing, Jilin, Jiangxi, Hubei and Guizhou Provinces showing the most significant growth. The number of injured people in 8 provinces and municipalities shows an increasing trend, with Jilin, Jiangxi, Hubei and Guizhou provinces showing significant growth. Conclusions Except for Hunan Province, it was difficult for the whole country and the rest provinces and municipalities to reach the SDGs target. According to the forecast results and the actual situation, a targeted scientific prevention and control strategy can be formulated. The measures taken by Hunan and Shanghai in traffic accident prevention and control were effective and worth learning.

OBJECTIVETo investigate whether injectable engineering heart tissue (EHT) can survive and improve heart function after transplantation into infarct area.

METHODSVentricular cardiomyocytes from 1-3 day-old Sprague-Dawley (SD) rats were isolated by using trypsin method, and then labeled and cultured. The left coronary of female SD rats was ligated to create a myocardial infarct model. Three weeks later, the qualified animals were randomized into four groups: EHT group (n = 12), which were transplanted with both cardiomyocytes and matrix; cell transplantation group (n = 12); matrix group (n = 12), control (n = 11). Four weeks after implantation, echocardiography and Langendorff model were used to assess heart function, and then the hearts were harvested for pathological examination. Meanwhile polymerase chain reaction (PCR) was performed to detect SRY gene on Y chromosome.

RESULTSThe grafted cells were identified in both EHT and cell transplantation group by either pathology or PCR. But in EHT group, transplanted cells formed more condensed tissue, and produced definite connected protein. Data of fraction shortness from echocardiography are showed as follows: EHT group, (22.82 +/- 3.44)%; cell transplantation group, (20.55 +/- 4.11)%, matrix group, (17.05 +/- 4.57)%; control, (19.80 +/- 3.98)% (P = 0.012). Langendorff examination revealed significant differences among four groups when left ventricular balloon volume was at the level of 0.06 ml, 0.08 ml and 0.10 ml, in which EHT group had the highest developed pressure and dp/dt.

CONCLUSIONIt is feasible to fabricate injectable EHT in vitro. The fabricated EHT could survive in the myocardial infarct area after transplantation in a rat model and improve heart function due to better histological configuration.

Animals ; Cell Transplantation ; methods ; Echocardiography ; Female ; Myocardial Infarction ; physiopathology ; surgery ; Myocytes, Cardiac ; transplantation ; Rats ; Rats, Sprague-Dawley ; Tissue Engineering

OBJECTIVETo investigate the regulatory effect of the nonsteroidal anti-inflammatory drug NS398 on the expression of the RECK gene in the animal model of prostate cancer.

METHODSNude mouse models of prostate cancer were divided into an experimental and a control group, the former fed with NS398 at 0.1 mg/g per day for 10, 20 and 30 days, while latter left without medication. All the mice were killed at 30 days, the mRNA expressions of RECK and MMP-9 in the tumor tissues measured by RT-PCR, and the protein level of RECK evaluated by Western blot.

RESULTSBoth the mRNA and protein expressions of RECK were increased, while the level of MMP-9 decreased, in an obviously time-dependent manner in the experimental group as compared with the control.

CONCLUSIONNS398 obviously inhibits the pathogenesis and metastasis of prostate cancer, which may be attributed to its induction of the expression of the RECK gene and suppression of the expression of MMP-9.

Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; GPI-Linked Proteins ; metabolism ; Gene Expression Regulation ; drug effects ; Humans ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Mice, Nude ; Nitrobenzenes ; pharmacology ; Sulfonamides ; pharmacology ; Tumor Cells, Cultured

OBJECTIVESTo investigate the regulative effect of the nonsteroidal anti-inflammatory drug NS398 on the RECK gene in the prostate carcinoma strain DU145.

METHODSDU145 was treated with various concentrations of NS398 for 48 hours. The mRNA level was measured by RT PCR technique and the expression of the RECK protein determined by Western blot.

RESULTSThe mRNA level of the RECK gene was obviously higher, while the MMP9 level markedly lower in the treated group than in the control, and so was the expression of the RECK protein.

CONCLUSIONNS398 induces the expression of the RECK gene, which might be the mechanism of its anti-tumor effect.

Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Blotting, Western ; Cattle ; Cell Line, Tumor ; GPI-Linked Proteins ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Male ; Membrane Glycoproteins ; biosynthesis ; genetics ; Nitrobenzenes ; pharmacology ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Sulfonamides ; pharmacology

OBJECTIVETo investigate the expression and significance of RECK gene and MMP-9 in prostate cell strains such as BPH-1, DU45, LNCaP and PC-3.

METHODSThe expression of mRNA of RECK and MMP-9 was measured by RT-PCR and RECK protein was evaluated by Western blot.

RESULTSThe mRNA level of the RECK gene in the prostate carcinoma cell strains, such as DU45, LNCaP and PC-3, was lower than that in the benign prostate hyperplasia cell line BPH-1, while MMP-9 had a higher expression. The protein level of RECK in DU-45, LNCaP and PC-3 was lower than that in the BPH-1.

CONCLUSIONThe RECK gene is supposed to be a kind of tumor suppressor gene, which may act by inhibiting the activity of MMP-9.

Blotting, Western ; Cell Line, Tumor ; GPI-Linked Proteins ; Humans ; Male ; Matrix Metalloproteinase 9 ; biosynthesis ; genetics ; Membrane Glycoproteins ; biosynthesis ; genetics ; Prostatic Neoplasms ; metabolism ; pathology ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction