Objective: To observe the expression of the G-protein-coupled-receptor 14 (GPR14), urotensinⅡreceptor, in the cardiovascular system and brain of SD rats. Methods: Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the GPR14 mRNA. Results: In cardiovascular system, GPR14 mRNA was detected in the left ventricle, left atrium, thoratic aorta and carotid aorta. The highest level of expression was found in the left ventricle. In the brain, GPR14 mRNA was detected in cortex, hippocampus, hypothalamus and cerebellum, and higher level of expression was found in the cerebellum. Conclusion: GPR14 mRNA expression is found in the cardiovascular and neural tissues of tested rat, suggesting that urotensinⅡ may play an important role in cardiovasculature and central nervous activity.

BACKGROUND:Osteoblasts with high purity and activity are essential for bone metabolism research. OBJECTIVE:To explore a simple and effective culturing method of primary osteoblasts. METHODS:Osteoblasts were isolated from the parietal and frontal bones of newborn Sprague-Dawley rats using trypsin and collagenase digestion and tissue explant method. The morphology of osteoblasts was observed by inverted phase contrast microscope and transmission electron microscope;the cells was counted to draw the growth curve;the osteoblasts were identified by alkaline phosphatase BCIP/NBT staining and alizarin red staining. RESULTS AND CONCLUSION:The cells showed spindle, triangle or polygon shapes, having two or three protrusions. There were abundant mitochondria and endoplasmic reticulum under electron microscope, which presented the typical characteristics of osteoblasts. The cell growth was slow intially, accelerating at the 3rd day, and peaking at the 7th day. The cells were highly positive for alkaline phosphatase staining and were stained orangered through the alizarin red staining. To conclude, the cells isolated using enzymatic digestion combined with tissue explant method exhibit the typical characteristics and functions of osteoblasts, and this method is an ideal way to culture primary osteoblasts.

Objective · To investigate the difference in myocardial strain of left ventricle between obstructive hypertrophy cardiomyopathy (HCM) and nonobstructive HCM. Methods · Cardiac magnetic resonance imaging (MRI) exam was performed on 48 sequential enrolled patients with HCM (18 with obstructive HCM, and 30 with nonobstructive HCM), whose left ventricular ejection fractions (LVEF) were over 50%. Twenty-five healthy volunteers were examined as normal controls. Global longitudinal strain (GLS), global radial strain (GRS), global circumferentialstrain (GCS), LVEF, left ventricular end diastolic volume (LVEDV), left ventricularmass (LVM), left ventricular end diastolic volume index (LVEDVI), and left ventricular mass index (LVMI) were collected and compared. Radial strain, circumferential strain and peak radial displacement were also measured in medial segment of left ventricle according to American Heart Association (AHA) 17-segment model. Results · ① LVEF of the patients with obstructive HCM was bigger than those of nonobstructive HCM patients and control group (P<0.05). LVM and LVMI of the HCM groups were bigger than those of control group (P<0.01). ② Left ventricle GLS, GRS, and GCS significantly decreased in the patients with nonobstructive HCM compared to those with obstructive HCM (P<0.05). The three parameters of two HCM groups were significantly lower than those of healthy volunteers (P<0.05). ③ Compared with obstructive HCM patients,the segmental parameters of left ventricule, the medial segment circumferential strain and radial strain of nonobstructive HCM patients significantly decreased (P<0.05), and the two parameters of both HCM groups were lower than those in healthy volunteers. Compared with obstructive HCM patients and healthy volunteers, peak radial displacement of left ventricule medial segment in nonobstructive HCM witnessed a significant decrease, while no significant difference was observed between obstructive HCM patients and healthy volunteers. Conclusion · In the LVEF preserved HCM patients, the myocardial strain of left ventricle in nonobstructive HCM patients decrease significantly than that in obstructive HCM patients, which may result in the different clinical outcomes intwo types of HCM patients. It is suggested that the myocardial strain is more sensitive than ejection fraction in the evaluation of myocardial performance of HCM patients.

Pseudomonas aernginosa (PA) is one of the most universal pathogens in clinical diagnosis, and conventional detection assay has many disadvantages. In this research, a pair of specific primers and a TaqMan fluorescent probe were designed in the conservative region of ETA gene by the method of bioinformatics analysis, the detection method for PA was successfully developed. Different gradient concentrations of PA DNA and various pathogen DNA were amplified by fluorescence quantitative PCR (FQ-PCR) to confirm the specificity and sensitivity of the developed method. Results showed that the developed detection assay is more sensible and specific by comparison to the conventional FQ-PCR method, and it is valuable for research and application prospects.
ADP Ribose Transferases ; genetics ; Bacterial Toxins ; genetics ; DNA, Bacterial ; analysis ; Exotoxins ; genetics ; Fluorescent Dyes ; Fluorometry ; methods ; Polymerase Chain Reaction ; methods ; Pseudomonas aeruginosa ; genetics ; isolation & purification ; Sensitivity and Specificity ; Taq Polymerase ; Virulence Factors ; genetics

Objective To investigate the effect of Dexamethasone (Dex) on the proliferation and apoptosis of osteoblasts in vitro and to explore its underlying mechanism.Methods Osteoblasts were acquired by primary culturing from new born SD rats.The inverted microscope was used to observe the cellular appearance.The cells were identified by alkaline phosphatase staining and alizarin red staining.The third generation osteoblasts were divided into four groups.Cells were incubated with different concentrations (0,10-8 mol/L,10-7 mol/L,10-6 mol/L) of dexamethasone for 12 hours,24 and 48 hours.Cell Counting Kit-8 was performed to evaluate the inhibitory effect on cell proliferation.The apoptosis rate was analyzed by flow cytometry with Annexin Ⅴ-FITC/PI double staining.The fluorescence microscopy was used to test the nuclear alteration and the expression of caspase-3.Western blot assay was applied to detect the expression of Bcl-2,Bad,caspase-3 and phosphorylated Akt.One-Way analysis of variance was used to determine the difference between groups.LSD-t was used to compare the difference between any two groups.Results Com-pared with the control group,dexamethasone at dose of 10-8 mol/L,10-7 mol/L and 10-6 mol/L inhibited the proliferation of osteoblasts,most evidently in 48 hours (0.980±0.028 vs 1.143±0.017,t=5.454,P＜0.05;0.798±0.057 vs 1.143±0.017,t=1 1.555,P＜0.05;0.728±0.031 vs 1.143±0.017,t=13.908,P＜0.05).Dexamethasone at dose of 10-7 mol/L and 10-6 mol/L induced apoptosis of osteoblasts at 48 hours,showing significant difference compared with control group [(9.8± 2.6)％ vs (4.1±0.8)％,t=3.508,P＜0.05;(12.4±2.6)％ vs (4.1±0.8)％,t=5.140,P＜0.05].However,10-8 mol/L of dexamethasone had no apparent effect in inducing apoptosis of osteoblasts [(4.9±1.2)％ vs (4.1±0.8)％,t=0.470,P ＞0.05].The immunofluorescene staining result showed that the expression of caspase-3 protein was significantly increased in 10-7 mol/L and 10-6 mol/L dex group (t=4.320,8.475,P＜0.05).The Western blotting results showed that dexamethasoneat the concentration of 10-7 mol/L,10-6 mol/L could significantly increase the expression of Bad and caspase-3 and down-regulate the expression of Bcl-2 and p-Akt.The expression of Bcl-2 was markedly reduced by 53.8％,78.4％ (t=4.019,5.988;P＜0.05),The expression of p-Akt decreased by 37％,49.6％ (t=2.067,3.491;P＜0.05),the expression of Bad protein increased by 276.9％ and 334.8％ respectively (t=7.342,8.872;P＜0.05),the expression of caspase-3 protein were increased by 138.0％ and 193.9％ (t=5.510,7.750;P＜0.05).Conclusion Dexamethasone is capable of inhibiting the proliferation of osteoblast,as well as augmenting the apoptosis.The mechanism of this process is probably related to reduction of the level of Bcl-2 expressionand up-regulation the expression of Bad,caspase-3 with the effects of inhibiting the PI3K/Akt signaling pathway.

OBJECTIVE: To investigate the association of myocardial blood flow (MBF) quantified by dynamic computed tomography (CT) myocardial perfusion imaging (MPI) with troponin level and left ventricle (LV) function in patients with ST-segment elevated myocardial infarction (STEMI). MATERIALS AND METHODS: Thirty-five STEMI patients who successfully had undergone reperfusion treatment within 1 week of their infarction were consecutively enrolled. All patients were referred for dynamic CT-MPI. Serial high-sensitivity troponin T (hs-TnT) levels and left ventricular ejection fraction (LVEF) measured by echocardiography were recorded. Twenty-six patients with 427 segments were included for analysis. Various quantitative parameters derived from dynamic CT-MPI were analyzed to determine if there was a correlation between hs-TnT levels and LVEF on admission and again at the 6-month mark. RESULTS: The mean radiation dose for dynamic CT-MPI was 3.2 ± 1.1 mSv. Infarcted territories had significantly lower MBF (30.5 ± 7.4 mL/min/100 mL versus 73.4 ± 8.1 mL/min/100 mL, p < 0.001) and myocardial blood volume (MBV) (2.8 ± 0.9 mL/100 mL versus 4.2 ± 1.1 mL/100 mL, p = 0.044) compared with those of reference territories. MBF showed the best correlation with the level of peak hs-TnT (r = −0.682, p < 0.001), and MBV showed a moderate correlation with the level of peak hs-TnT (r = −0.437, p = 0.026); however, the other parameters did not show any significant correlation with hs-TnT levels. As for the association with LV function, only MBF was significantly correlated with LVEF at the time of admission (r = 0.469, p = 0.016) and at 6 months (r = 0.585, p = 0.001). CONCLUSION: MBF quantified by dynamic CT-MPI is significantly inversely correlated with the level of peak hs-TnT. In addition, patients with lower MBF tended to have impaired LV function at the time of their admission and at 6 months.
Blood Volume ; Echocardiography ; Heart Ventricles ; Humans ; Infarction ; Myocardial Infarction ; Myocardial Perfusion Imaging ; Reperfusion ; Stroke Volume ; Troponin T ; Troponin

ObjectiveTo evaluate the neurobiological characteristics of human histio-amniotic mesenchymal (hAMCs) and effect of hAMCs transplantation into the brain to treat Parkinson's disease(PD) modle mice.MethodsThe expressions of mesenchymal stem cells, neural stem cells, dopaminergic neurons and markers related to neurogenesis such as Vimentin, STRO-1, nestin, CD133, β-tubulin, TH, DAT, Ngn2 and mash-1 in hAMCs were evaluated through immunocytochemical stain; and the mRNA transcriptions of neural stem cell markers, Vimentin and nestin in hAMCs were detected by RT-PCR. The PD model was induced by MPTP(i.p.) in C57BL/6 mice transplanted with hAMCs into the right striatum. The therapeutical effect of hAMCs on PD mice was evaluated by spontaneous movement, rotating bar test and the immunohistochemistry of anti-human chondrosome and TH antibodies in striatum.ResultshAMCs induced by nerve cells culture medium, expressed mesenchymal stem cells, neural stem cells, dopaminergic neurons and other specific markers related to neurogenesis mentioned above. The frequency of spontaneous movement in PD mice was significantly increased(P<0-05), and the time of rotating bar was obviously prolonged(P<0-05) after transplantation with hAMCs.ConclusionhAMCs possess the characteristics of nerve cells after cultured in vitro and can significantly recover the damage of motor function induced by MPTP after transplantation into striatum in PD model mice.

Porcine deltacoronavirus (PDCoV) has been recently recognized as an emerging viral pathogen that causes diarrhea in newborn piglets. A total of 254 small intestinal or fecal samples collected from 10 provinces including Henan, Hunan, Zhejiang, Jiangxi, Anhui, Hebei, Heilongjiang, Jiangsu, Shandong and Shanghai between 2014 and 2015, were screened by quantitative RT-PCR targeting the viral M gene. Eleven PDCoV positive samples were identified with a total positive rate of 4.33%. An indirect enzyme-linked immunosorbent assay (ELISA) was developed based on the recombinant S1 protein of PDCoV. This assay was used to test 609 serum samples of pigs with diarrhea symptoms collected from 10 provinces between 2015 and 2016. The positive rate of PDCoV antibody was 44.17% (269/609). The two methods can be used to monitor the PDCoV epidemiology in the levels of PDCoV specific RNA or antibody, helping better prevent and control PDCoV.

Objective To explore the value of preoperative detection of soluble fragments of cytokeratin-19 (CYFRA21-1), carcinoembryonic antigen (CEA), and postoperative detection of nuclear proliferation associated antigen Ki67 in prognostic evaluation of non-small cell lung cancer patients. Methods The clinicopathological data and follow-up results of patients with non-small cell lung cancer treated in the Department of Thoracic Surgery of the First Affiliated Hospital of Xiamen University in 2017 were collected. CYFRA21-1>3.39 ng/mL was defined as positive, and CEA>5 ng/mL was defined as positive. The receiver operating characteristic curve (ROC curve) of Ki67 expression level was drawn. The maximum area under the curve (AUC) was the cutoff value of Ki67 expression level, and the Ki67 expression level greater than its cutoff value was defined as positive. Cox regression analysis was used to determine the independent risk factors for poor prognosis in patients with non-small cell lung cancer. Results Finally 248 patients were collected, including 125 males and 123 females, with a median age of 61 years (ranging from 30 to 81 years) at the time of surgery. Univariate analysis showed that positive CYFRA21-1, high expression of Ki67, positive CEA, age≥60 years at operation, distant metastasis, lymph node metastasis, maximum tumor diameter>3 cm, and TNM stage Ⅲ were associated with poor prognosis in patients with non-small cell lung cancer. When combined detection of preoperative tumor markers and postoperative Ki67, the prognosis of all negative patients was the best, and that of all positive patients was the worst. Cox regression analysis showed that positive CEA+positive CYFRA21-1+high expression of Ki67 was an independent risk factor for poor prognosis in patients with non-small cell lung cancer (P<0.05). Conclusion The combined detection of preoperative serum CYFRA21-1, CEA, and postoperative Ki67 has important value in evaluating the prognosis of non-small cell lung cancer patients.

Objective    To investigate the clinical effect of three-port Da Vinci robot-assisted radical resection of lung cancer. Methods    The clinical data of patients who underwent Da Vinci robot-assisted radical resection of lung cancer in the Second Department of Thoracic Surgery, the First Affiliated Hospital of Xiamen University from April 2021 to March 2022 were retrospectively analyzed. According to the number of surgical ports, they were divided into two groups: a three-port group (three-port Da Vinci robot-assisted radical resection of lung cancer), and a four-port group (traditional Da Vinci robot-assisted radical resection of lung cancer). The operation time, intraoperative bleeding, lymphadenectomy, total thoracic drainage, extubation time, postoperative complications and postoperative pain of the two groups were compared and analyzed. Results    A total of 58 patients were included, including 19 males and 39 females, aged 31-79 years. There were 21 patients in the three-port group, and 37 patients in the four-port group. The visual analogue scores on the first and third day after the operation were 4.33±1.20 points and 2.24±0.77 points in the three-port group, and 5.11±1.22 points and 2.78±1.06 points in the four-port group, and there were statistical differences between the two groups (P<0.05). There was no significant difference between the two groups in terms of operation time, intraoperative bleeding, lymph node dissection, postoperative thoracic drainage, time of thoracic tube insertion or postoperative complications (P>0.05). Conclusion    Three-port Da Vinci robot-assisted radical resection of lung cancer can reduce the postoperative pain without increasing the operation difficulty and complications, and can be widely used in  the clinical practice.