Objective: To observe morphological changes of epithelial-mesenchymal transition in the early stage of mice renal interstitial fibrosis. Methods: Renal interstitial fibrosis was induced by unilateral ureteral obstruction(UUO) in mice. Histological and immunohistochemical methods were used to analyze pathological changes and α-SMA expression in renal tissue.Argentum hexamethylenamine staining and transmission electron microscopy were used to observe changes of the renal tubule basement membrane. Gelatin zymographic analysis was used to observe the expression of MMP2 and MMP9 in renal tissue.Results:The mice suffered from renal interstitial fibrosis were identified by histological analysis and α-SMA positive cells in renal tissue. Argentum hexamethylenamine staining and transmission electron-microscopy showed that the renal tubule basement membrane disrupted locally and renal tubule epithelial cells invaded into the renal interstitium in the early stage of renal interstitial fibrosis. Gelatin zymographic analysis showed that the expression of MMP2 and MMP9 was increased transitorily in the early stage of renal interstitial fibrosis. Conclusion: Renal tubule basement membrane disruption, renal tubule epithelial cells invasion into the renal interstitium, and the expression of MMP2 and MMP9 are involved in the development of renal interstitial fibrosis.

ObjectiveTo investigate the correlation between serum C-reactive protein(CRP),tumor necrosis factor-alpha(TNF-α) and atherosclerosis in patients with subclinical hypothyroidism.Methods Seventy-five patients with subclinical hypothyroidism were divided into two groups according to the level of TSH:mild group (42 cases) with TSH between 5.5 and 10.0 mU/L and severe group (33 cases) with TSH ＞10.0 mU/L.Thirty healthy physical examinees or volunteers were selected as control group.TG,LDL-C,HDL-C,TSH,FT4,FT3,CRP,TNF-α and carotid intima-media thickness(CIMT) of all cases were detected and measured.ResultsCIMT of mild,severe and control groups were(0.88 ± 0.20),( 1.12 ±0.21 ),(0.62 ±0.21 ) mm,respectively.CIMT of mild and severe groups were both significantly higher than that of control group,CIMT of severe group was significantly higher than that of mild group (P ＜ 0.01 ).There were no significant differences of CRP and TNF- α among three groups (P ＞ 0.05 ).LDL-C of mild and severe groups were significantly higher than that of control group [ (3.22 ± 0.37),(3.49 ± 0.38 ) mmol/L vs.(2.48 ±0.41 )mmol/L,P ＜0.01 ].HDL-C and TG between mild and control groups had no statistical significance (P ＞0.05).Compared with control and mild groups,HDL-C of severe group decreased [(0.92 ±0.10)mmol/L vs.( 1.21 ± 0.14),( 1.17 ± 0.11 ) mmol/L] and TG increased [ ( 1.50 ± 0.49) mmol/L vs.( 1.11 ±0.53),(1.27 ±0.47) mmol/L](P＜ 0.01 or ＜ 0.05).The levels of TG,LDL-C,TSH,CRP and TNF-α had positive correlation with CIMT (r =0.52,0.37,0.48,0.39,0.45;P ＜0.05 or ＜0.01 ).FT4 was negatively correlated with CIMT (r=-0.24,P＜0.05).HDL-C had no relation with CIMT (r =0.06,P ＞0.05).ConclusionsThere are many abnormalities such as metabolism disorder of blood lipid in patients with subclinical hypothyroidism.The risk of atherosclerosis increases in these patients and inflammatory reaction may not be an important factor of subclinical hypothyroidism complicated with atherosclerosis.

Objective To study the expression and significance of multidrug resistance gene (MDR1) in children with refractory epilepsy (RE). Methods Children with RE (n = 30), non-RE (n = 30) and healthy children (n=30) were collected. The expression of MDR1-mRNA in peripheral blood was analyzed by fluorescence quantitative PCR. The relationship of MDRI-mRNA with epileptic frequency and numbers of antiepileptic drugs (AEDs) were observed. Results The expression of MDR1 in RE group obviously increased when compared with that of non-RE group and healthy group (P < 0.01, P < 0.01) ; MDR1 expression was more among patients with high frequent epilepsy than patients with low frequent epilepsy (P < 0.01) ; more in patients administered with four kinds of AEDs than those with two or three kinds of AEDs (P < 0.01). Concinsions MDR1 overexpression in blood of children with RE may be linked to drug-resistant mechanism of RE. It might be used as a clinical indicator of RE.

Objective To study the application of contrast-enhanced ultrasonography (CEUS) in the evaluation of the stability of carotid atherosclerotic plaque (CAP).Methods 162 patients with CAP were selected as study group.Meanwhile,34 patients with carotid artery strong echo plaques were selected as control group.The color doppler ultrasound was used to observe the CAP.Results The proportions of lipid type,fiber type,calcification and ulcer plaque in the study group were 21.60%,33.33%,34.57% and 10.37%,which were higher than those of the control group (5.88%,2.94%,2.94% and 2.94%),the differences were statistically significant (χ2=4.537,12.859,13.629 and 3.855,all P<0.05).There were 75 patients of soft plaques,36 patients with mixed plaques,51 patients with hard plaque in 162 patients.The new blood vessels classification in soft plaque group (36.00%,45.33% and 10.33%) were higher than the mixed plaque (30.56%,41.67% and 8.33%) and hard plaque group (31.37%,13.72% and 7.84%).The peak intensity (-86.41±7.81) %,tmax (8.34±1.62)s,mean transit time (24.18±8.67)s in the soft plaque group were significantly lower than the mixed plaque [(-100.73±6.52)%,(9.79±2.14)s and (28.93±9.11)s] and hard plaque patients [(-104.14±6.15)%,(10.23±2.33)s and (30.07±9.48)s],the differences were statistically significant (t=9.518,6.966,2.658,13.592,5.374 and 3.064,all P<0.05),but there were no statistically significant differences between mixed plaque and hard plaque (all P>0.05).The plaque diameter (4.13±0.75)mm diagnosed by CEUS was significantly larger than that of conventional ultrasound [(3.62±1.14)mm],the difference was statistically significant (t=4.757,P=0.000).Conclusion The CEUS can qualitatively detect the atherosclerotic plaque angiogenesis,can quantitatively assess plaque,evaluate the stability of the plaques,and the sensitivity is high.

Objective To evaluate the effect of nimodipine combined with 7.5％ hypertonic saline (HS) on postoperative cognitive function in aged rats.Methods Ninety-six healthy male Wistar rats,aged 18 months,weighing 450-500 g,were assigned into 4 groups (n=24 each) using a random number table:splenectomy group (group S),nimodipine group (group N),group HS and nimodipine plus HS group (group N+HS).Nimodipine 1 mg/kg was intraperitoneally injected in group N.In group HS,7.5％ HS 4 ml/kg was injected via the caudal vein.The equal volume of normal saline was injected intraperitoneally or via the caudal vein in group S.Splenectomy was performed under sevoflurane anesthesia at 30 min after the end of administration.On 1 day before operation and 3 and 7 days after operation,Morris water maze test was performed,and blood sainples from the caudal vein were simultaneously collected for determination of the concentrations of serum S100β protein and neuron-specific enolase (NSE) by enzyme-linked immunosorbent assay.Results Compared with group S,the frequency of crossing the original platform was significantly increased,the escape latency was shortened,and the concentrations of serum S100β protein and NSE were decreased at each time point after operation in N,HS and N+HS groups (P＜0.05).Compared with group N or group HS,the frequency of crossing the original platform was significantly increased,the escape latency was shortened,and the concentrations of serum S100β protein and NSE were decreased at each time point after operation in group N+HS (P＜0.05).Conclusion Nimodipine combined with 7.5％ HS exerts better efficacy than either alone in improving postoperative cognitive function in aged rats.

OBJECTIVEConstructing a plasmid containing the shRNA of luciferase to suppress the expression of luciferase in BHK-21 cell.

METHODSA 334 bp human U6 snRNA promoter was amplified from human genomic DNA by PCR and ligated to a 21 bp reverse repeated motif of luciferase target sequence with 9 bp spacer and AAV plasmid pSNAV. The recombinant pSNAV/U6/Luc plasmid cotransfected with pMAMneoLuc or transfected luciferase cell line to detect the effect of luciferase expression separately.

RESULTSpSNAV/U6/luc suppresses the luciferase expression from pMAMneoLuc by 50% and luciferase cell line by 70%.

CONCLUSIONSThe results showed that the short hairpin RNA of luciferase can efficiently suppress its expression in BHK-21.

Cell Line ; Gene Expression ; Genetic Vectors ; Humans ; Luciferases ; biosynthesis ; genetics ; Plasmids ; genetics ; Promoter Regions, Genetic ; RNA ; genetics ; Transfection

Objective:To explor the effects of mentalization-based family therapy (MBFT) on the emotions of anxiety and depression, reflective functions, and related brain regions of adolescents with depression.Methods:Thirty-two adolescent patients with depression were selected in Beijing Huilongguan Hospital.Among them, fifteen patients treated with conventional drugs were defined as the control group, the other seventeen patients served as the observation group.On the basis of routine drug treatment, patients in the observation group were treated with 1.5 h MBFT once a week, for a total of 3 months.Among them, 10 patients underwent resting functional magnetic resonance imaging(rs-fMRI) scans before and after the intervention.SPSS 23.0 was used for statistical analysis of scale evaluation data by using chi-square test and repeated-measure analysis of variance, and pairwise Pearson correlation was used to construct a brain network matrix for the MRI results.The patient's brain network matrix was put into Gretna to calculate, and then use repeated measures analysis of variance and t test for judgment. Results:(1)Repeated measurement analysis of variance showed, over time and group, before and after intervention, the group×time interactions of patients' reflective function, anxiety, depression were statistically significant ( F=5.113, 9.350, 5.264, all P<0.05). Further simple effect analysis showed, in the observation group, there were statistically significant differences in patients' anxiety ((55.09±14.35) vs (30.58±3.62)), depression ((61.58±16.81) vs (31.00±3.48)) before and after intervention (all P<0.01). There was no significant difference in reflective function ((8.71±0.51) vs (8.87±0.50) ) in the observation group before and after intervention( P> 0.05). The reflective function((8.92±0.33) vs (8.73±0.35)) and anxiety((50.67±13.88) vs (45.78±12.89)) scores of the control group were not statistically significant different before and after intervention(all P>0.05), and the depression scores of the control group before and after inter-vention((69.33±9.11) vs (50.94±7.99)) were statistically significant different( P<0.01). (2)rs-fMRI showed that, using the posterior parietal lobe and the medial prefrontal lobe as seed points, the group×time interactions with right inferior frontal gyrus, right fusiform gyrus, right superior temporal gyrus, left middle temporal gyrus and right superior frontal gyrus were statistically significant ( F=29.56, 20.63, 8.69, 9.49, 8.62, all P<0.05). The further simple effect analysis showed in the observation group, the pre-test was less than the post-test, and the difference was statistically significant( P<0.05). In the control group, there were not statistically significant differences in the functional connection strength before and after the test(all P>0.05). (3)The brain network analysis of 264 regions of interest (ROIs) in the whole brain showed that the number of modules in the observation group was less than that in the control group, and the difference was statistically significant ( t=-2.64, P=0.046). Conclusion:MBFT can improve the anxiety and depression of adolescent patients with depression.It enhances the functional link between mentalization related brain regions and the whole brain, and enhances the activity of the whole brain link to become more integrated.

Objective　To investigate the effects of free radicals (FRs) and amyloid β protein 1-40 (Aβ1-40) on the functions of expressed neurotransmitter receptors (NRs) in Xenopus oocytes. Methods　Total RNA and messenger RNA (mRNA) was prepared from 3-month-old Wistar rat brain tissues with Promega kits and microinjected into maturated Xenopus oocytes (stages Ⅴ-Ⅵ) with 50nl (50ng) for each oocyte. The microinjected oocytes were incubated with modified Bath's solution at 19.0℃±1.0℃ for receptor expression and their currents were recorded with double electrode voltage clamp technique. Superoxide anion free radicals (SAFRs) were produced via a reaction system (HPX/XO) with hypoxanthine (HPX, 0.05mol/L) and xanthine oxidase (XO, 0.1U/L). In order to observe the effects of Aβ and SAFRs on the expressed glutamate receptor, HPX/XO and Aβ1-40 were added to incubation solution at 12h, 24h and 96h before recording. Results　The results showed that the oocytes expressed functional NRs originating from rat brain tissues. These NRs included muscarinic acetylcholine (mACh), glutamate (Glu), dopamine (DA), serotonin (5-HT) and γ-aminobutyric acid (GABA). The current characteristics of expressed receptors were inward currents carried by chloride ion with their equibrilium potentials close to -22mV. The extent of effect on the current of expressed glutamate receptor from rat brain was different among different Aβ concentrations and incubation times. Aβ1-40 at a concentration of 20nmol/L had little effect on the currents of expressed rat brain glutamate receptors up to 24h of incubation period; but the currents of glutamate receptor were significantly decreased (25% off, P<0.01) in the treatment of 60nmol/L Aβ1-40 over 24h. Moreover, when 20nmol/L Aβ1-40 was co-incubated over 12h with SAFRs produced by the reaction system of HPX/XO, it was found that the currents of expressed rat brain glutamate receptors had been changed markedly. When the oocytes were co-treated with 60nmol/L Aβ1-40 and SAFRs over a period of 12h, the currents of glutamate receptor significantly decreased (21% off, P<0.05), and the decreased percentage reached 52% over 24h co-treatment with 60nmol/L Aβ1-40 and SAFRs. In addition, vitamin E had a partial effect against this inhibitory effect. Conclusion　The results suggest that Aβ has a kind of inhibitory effect upon the current of the glutamate receptor, similar to the effects of free radicals. The effects can be antagonized by vitamin E. These imply that Aβ may play a role via inhibiting receptor function in the pathophysiology of Alzheimer's disease.

Objective:To investigate the correlation between cyclooxygenase-2(COX-2), estrogen receptor (ER), progesterone receptor(PR) and the formation of endometrial polyps and the increased malignant rate of postmenopausal endometrial polyps.Methods:From January 2018 to January 2019, a total of 160 postmenopausal women treated in the People's Hospital of Changxing County were selected and divided into endometrial polyps group(83 cases) and oothectomy group(42 cases), normal menopause group(35 cases) according to different diseases.The levels of COX-2, ER, PR in the three groups were measured.The expression of COX-2 in mesenchymal cells and glandular epithelial cells, the expression of ER in mesenchymal cells and glandular epithelial cells, the expression of PR in mesenchymal cells and glandular epithelial cells in the three groups were compared, and the correlation between COX-2, ER, PR and increased malignant rate of endometrial polyps was analyzed.Results:The positive expression rates of COX-2 in mesenchymal cells and glandular epithelial cells in the endometrial polyps group(16.9%, 30.1%) were higher than those in the normal menopause group(0.0%, 11.4%) and the oothectomy group(4.8%, 7.1%)(χ 2=4.568, 5.806, all P<0.05 ). There were no statistically significant differences in the expression of ER in mesenchymal cells and glandular epithelial cells among the three groups(χ 2=1.333, 1.412, all P>0.05). The expression of PR in mesenchymal cells and glandular epithelial cells in the endometrial polyps group was lower than that in the normal menopause group and the oothectomy group(χ 2=4.890, 5.022, all P<0.05). COX-2 was positively correlated with the increase in malignant rate of endometrial polyps( r=4.335, P<0.05), PR was negatively correlated with an increase in the malignant rate of endometrial polyps( r=-4.256, P<0.05), and ER had no significant correlation with an increase in the malignant rate of endometrial polyps( r=1.203, P>0.05). Conclusion:COX-2 and PR are significantly related to the formation of endometrial polyps and the increase in malignant rate of postmenopausal endometrial polyps, but there is no significant expression of ER.Therefore, the detection of COX-2 and PR levels is beneficial to provide a certain scientific basis for clinical treatment.

Poly-adenosine diphosphate ribose polymerases (PARPs) play an important role in DNA repair and apoptosis.Among them, mono-(ADP-ribosyl) transferase (MARTs) can regulate various cell reactions by catalyzing and transferring single ADP-ribose.Most MARTs are highly expressed in cancers, which is closely related to the occurrence and progression of cancers.This review introduces the MARTs that are highly expressed in cancers, classifies them according to the differences of their structural domains, and reviews their known mechanism, their close relationship with cancers, their potential value in cancer therapy and the research progress of corresponding inhibitors.These targets are expected to provide new research ideas for cancer therapy in the era of precision medicine.