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Objective:To investigate the effects of various concentrations of recombinant human WISP2 protein(WISP2)on lipid metabolism in HepG2 cells.Methods:HepG2 cells were treated with different concentrations(0, 0.4, 1 and 2 μg/L)of recombinant human WISP2 for 48 hours.Cell viability was detected by Cell-Titer, and enzymatic hydrolysis methods were used to measure intracellular triacylglycerol(TG)and total cholesterol(TC)levels.The mRNA expression was detected by quantitative real-time reverse transcription-PCR(RT-qPCR)and protein expression in HepG2 cells was detected by western blot.Results:Compared with the control group, the WISP2 groups treated with various concentration did not significantly reduce the viability of HepG2 cells.TG and TC in HepG2 cells were significantly increased by recombinant human WISP2 treatment(all P<0.05).The concentrations of TG in the 0.4, 1 and 2 μg/L recombinant human WISP2-treated groups were 1.254±0.039, 1.216±0.028 and 1.174±0.014)times the concentration in the untreated group, respectively( F=6.791, P=0.006).The concentration of TC in the untreated group was 1.264±0.057, 1.394±0.101 and 1.392±0.077), respectively, times the concentration in each of the treated groups( F=7.045, P=0.005).Further experiments found that the mRNA expression of sterol regulatory element binding protein 1(SREBP1), 3-hydroxy-3-methylglutaryl coenzyme A reductase(HMGCR), acetyl-CoA carboxylase(ACC), type 2 diacylglycerol acyltransferase(DGAT2)and the protein expression of SREBP1, ACC and fatty acid synthase(FAS)were significantly increased in the recombinant human WISP2-treated groups, compared with the control group(all P<0.05).However, the expression of lipid transporters such as the low-density lipoprotein receptor(LDLR), ApoB and ApoE and adipose triglyceride lipase(ATGL), a key lipolysis protein, was not significantly affected. Conclusions:Human recombinant WISP2 protein increases lipid levels in hepatocytes and the key underlying mechanisms may be through promoting lipid synthesis.
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Objective:To investigate the risk factors for cerebral injury in survivors of twin-to-twin transfusion syndrome (TTTS) after fetoscopic laser occlusion of chorioangiopagous vessels(FLOC) and to analyze the neurodevelopmental outcomes at 12 months of corrected age.Methods:A total of 136 cases of TTTS receiving FLOC in the Third Affiliated Hospital of Zhengzhou University from May 2018 to August 2021 were retrospectively selected as the FLOC group, and the survivors were followed up. Neurological development at 12 months of corrected age was assessed using the Griffiths mental development scales-Chinese (GDS-C) from five dimensions with locomotor, personal-social, hearing and language, hand-eye coordination and performance subscales. Eighty-eight fetuses of TTTS pregnancies receiving expectant treatment or amniotic fluid reduction were selected as the non-FLOC group. The perinatal mortality and the incidence of cerebral injury in the two groups were compared, as well as the incidence of cerebral injury between patients undergoing Solomon surgery and selective laser surgery in the FLOC group. Generalized estimating equations were used to analyze the risk factors for neonatal cerebral injury after FLOC and the factors influencing general developmental quotient score at the corrected age of 12 months. Chi-square test, t-test, and Mann-Whitney U test were used for statistical analysis. Results:(1) The perinatal mortality rate in the FLOC group was lower than that in the non-FLOC group [14.7% (20/136) vs 26.1% (23/88), χ 2=4.50, P=0.034]. There was no statistical significance in the incidence of neonatal cerebral injury between the two groups [18.7% (23/123) vs 21.8% (17/78), χ 2=0.29, P=0.592], but the incidence of severe cerebral injury in the FLOC group was lower than that in the non-FLOC group [6.5% (8/123) vs 15.4% (12/78), χ 2=4.20, P=0.040]. (2) In the FLOC group, there was no significant difference in the incidence of cerebral injury between donors and recipients, or between Solomon surgery and selective laser surgery [16.4% (10/61) vs 21.0% (13/62), χ 2=0.42; 20.0% (9/45) vs 17.9% (14/78), χ 2=0.08; both P>0.05]. (3) Multivariate analysis showed that neonatal asphyxia ( OR=7.04, 95% CI: 1.45-34.20, P=0.016) and higher preoperative TTTS stage ( OR=2.05, 95% CI: 1.10-3.82, P=0.023) were risk factors for neonatal cerebral injury. (4) Fifty-two cases were successfully followed up at the corrected age of 12 months, and the incidence of developmental delay in at least one dimension was 34.6% (18/52). Developmental delay was mainly manifested in locomotor skills and language, accounting for 26.9% (14/52) and 11.5% (6/52). No significant difference in Z value was found between recipients and donors in each dimension (all P>0.05). Solomon surgery, larger gestational age at operation and low birth weight were related to low general developmental quotient score (95% CI:－11.71 to－0.23，－1.99 to－0.47，0.00-0.01，respectively，all P<0.05). Conclusions:The occurrence of cerebral injury in TTTS survivors after FLOC is related to preoperative TTTS staging and intrapartum neonatal asphyxia. Neurodevelopment of survivors is related to birth weight and gestational age at surgery, and there is a higher incidence of mild developmental delay at corrected age of 12 months.
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Objective:To explore the significance of proactive healthy living index(PHLI)to provide a reference for the elderly to maintain function through healthy daily life.Methods:The functional status of the elderly was defined based on gait speed and cognitive function.Logistic regression model was used to select the daily life indicators which could be the risk factors for the functional status, and a nomogram was graphed for its visualization.PHLI was formed according to the result of the nomogram.Receiver operating characteristic curve(ROC)was plotted according to the functional status of the elderly, and the cut-off value of PHLI was determined based on the principle of the maximum sum of sensitivity and specificity.Results:A total of 4 971 participants with an average age of (70.9±7.5)years were included in the cross-sectional study, of whom 2 649 cases(53.3%)maintained normal function.The Logistic regression analysis showed that the housework had the most significant effect on the functional status of the elderly, followed by positive psychology( OR=2.04, 1.92; both P<0.001), increasing the likelihood of maintaining normal function by 104% and 92%, respectively.Regular life patterns, adequate protein intake, sleep quality and so on also influenced functional status of the elderly(all P<0.05). The PHLI median score of the elderly was 7.3(5.9, 8.3)points.The area under the curve(AUC)was 0.70(95% CI: 0.68-0.71), and the cut off value of PHLI was 6.8 points. Conclusions:The PHLI is associated with the maintenance of functional capacity in the elderly, which can be applied as a simple tool to provide reference for old people to keep functional health through living a proactive healthy daily life.
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Objective: To explore the clinical manifestations and genetic characteristics of patients with epilepsy and episodic ataxia caused by SCN2A gene variation. Methods: The clinical data of seizure manifestation, imaging examination and genetic results of 5 patients with epilepsy and (or) episodic ataxia because of SCN2A gene variation admitted to the Department of Pediatrics, the Third Affiliated Hospital of Zhengzhou University from July 2017 to January 2021 were analyzed retrospectively. Results: Among 5 patients, 4 were female and 1 was male. The onset age of epilepsy ranged from 4 days to 8 months. There were 2 cases of benign neonatal or infantile epilepsy and 3 cases of epileptic encephalopathy, in whom 1 case had development retardation,1 case transformed from West syndrome to infantile spasm and another one transformed from infantile spasm to Lennox-Gastaut syndrome. One case of benign neonatal-infantile epilepsy was characterized by neonatal onset seizures and episodic ataxia developed at the age of 78 months. Electroencephalograms at first visit of 5 cases showed that 2 cases were normal, 1 case had focal epileptic discharge, and 2 cases had multi-focal abnormal discharge with peak arrhythmia. The brain magnetic resonance imaging (MRI) of 3 cases were nomal, 1 case was abnormal (brain atrophy with decreased white matter) and the results of 1 case was unknown. The follow-up time ranged from 17 months to 89 months. Four cases of epilepsy were controlled and 1 case died at 2 years of age. Two cases had normal intelligence and motor development, 2 had moderate to severe intelligence retardation and motor critical state, and 1 had moderate to severe intelligence and motor development retardation. SCN2A gene variations were identified in all cases. There were 4 missense variations and 1 frameshift variation. Three variations had not been reported so far, including c.4906A>G,c.3643G>T,c.638delT. Conclusions: Variations in SCN2A gene can cause benign neonatal or infantile epilepsy and epileptic encephalopathy. Some children develop episodic ataxia with growing age. The variation of SCN2A gene is mainly missense variation.
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Child , Female , Humans , Infant , Infant, Newborn , Male , Ataxia/genetics , Electroencephalography , Epilepsy/genetics , Mutation , /genetics , Retrospective Studies , Spasms, Infantile/geneticsABSTRACT
Objective:To investigate the effects and potential mechanisms of resveratrol on obesity in elderly mice.Methods:In this study, 3 groups were randomly formed for 32-week-old mice and for 48-week-old mice.The normal diet group received regular chow and 0.3 ml saline by gavage once a day, the high-fat diet group received a high-fat diet(containing 21% fat and 1.25% cholesterol)and 0.3 ml saline once a day, and the high-fat diet plus resveratrol group received a high-fat diet and resveratrol(22.4 mg/kg, dispersed in 0.3 ml saline)by gavage once a day.After 12 weeks, body weight and adipose tissues were measured.Plasma leptin concentrations were determined by an enzyme-linked immunosorbent assay(ELISA), and values for hypertrophic obesity-related indexes of mice were obtained by quantitative real-time PCR.Results:The body weight and the proportion of subcutaneous fat tissues were lower in the high-fat diet plus resveratrol group than in the high-fat diet group[(34.43±3.23)g vs.(53.16±2.16)g, (3.21±1.58)% vs.(4.86±0.64)%, P<0.01], and were similar to those in the normal diet group.Resveratrol had a more obvious inhibitory effect on leptin in elderly mice than in middle-aged mice.In elderly mice, the plasma leptin concentration was lower in the high-fat diet plus resveratrol group than in the high-fat diet group[(0.015±0.009)g/L vs.(0.100±0.027)g/L]and the normal diet group( F=19.85, P=0.001), and it was similar to that in the middle-aged mice on a normal diet.Resveratrol significantly increased the expression of peroxisome proliferator-activated receptor gamma(PPARγ)and glucose transporter 4(GLUT4)and reduced the expression of tumor necrosis factor-α(TNF-α)( F=10.79, 9.31 and 7.02, P=0.003, 0.006 and 0.010). Conclusions:Resveratrol can significantly improve hypertrophic obesity in elderly mice, and the inhibition of leptin secretion and up-regulation of PPARγ may be the key mechanisms.
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Objective:To explore the relationship between doing housework and gait speed(GS)in order to identify ways to maintain physical capability in elderly people.Methods:A nationwide cross-sectional survey was conducted from Jan.2011 to Dec.2011.Subjects in this survey, with a sample of 2 519 males and 2 541 females aged ≥60 years, came from six regions across the country and satisfactorily completed a GS test.A GS in the lower 20% of the reference population(<0.7 m/s)was used as the cut-off point to define slow GS(SGS). Housework was defined as activities such as house cleaning, shopping, cooking and taking care of grandchildren and a response of"often"(≥5 d/week)was considered as doing housework.Propensity Score Matching(PSM)was performed to select comparable sub-groups of the study population to evaluate the association between housework and GS.Results:There were 3 290 subjects(1 645 pairs)selected by PSM.Logistic regression analysis showed that doing housework was inversely associated with SGS, and housework activities reduced the risk of SGS by 60%( OR=0.40, 95% CI: 0.35-0.46, P<0.01). Conclusions:Elderly people who conduct housework often are more likely to maintain GS and therefore should be encouraged to engage in daily activities to help maintain adequate physical performance.
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Objective:To investigate the effects of lithium chloride on vascular smooth muscle cell calcification and the potential underlying mechanisms.Methods:Human aortic smooth muscle cells were cultured in vitro, and a smooth muscle cell calcification model was established by using a calcification medium(the concentration of inorganic phosphorus was 3 mmol/L). Cells in the drug treatment group were pretreated with lithium chloride(10 mmol/L)for 4 hours and then with inorganic phosphorus at 3 mmol/L.After several days in culture, calcium deposition in cells was measured by alizarin red S staining.The secretion of extracellular pyrophosphate was detected by measuring nicotinamide adenine dinucleotide hydrogen(NADH) consumption of pyrophosphate-coupled enzyme reactions, which were monitored spectrophotometrically at 340 nm.Real-time PCR and Western blotting were used to detect mRNA and protein expression of the human progressive ankylosis( ankh)gene.Human aortic smooth muscle cells were infected with the scramble control lentivirus and the sh- ankh lentivirus, respectively, to establish the control cell group and the ankh knockdown cell group.The effects of ankh knockdown on cell calcification were examined. Results:The calcification level in vascular smooth muscle cells increased in the high inorganic phosphorus group, compared with the control group[(65.00±2.11)ng/g vs.(12.39±0.38)ng/g, P<0.01)]. Compared with the high-phosphorus control group, lithium chloride evidently inhibited high phosphate-induced vascular smooth muscle cell calcification[(24.92±1.87)ng/g vs.(60.94±4.51)ng/g, P<0.01)]. Lithium chloride pretreatment clearly increased extracellular pyrophosphate levels under unstimulated conditions[(51.70±7.26)×10 -3mmol/g vs.(28.71±2.55)×10 -3mmol/g( P<0.01)]and under high phosphorus stimulation[(34.35±4.27)×10 -3mmol/g vs.(20.89±4.93)×10 -3mmol/g( P<0.05)], and increased the expression of ankh as well( P<0.01). In addition, ankh knockdown markedly enhanced the extent of inorganic phosphorus-induced vascular smooth muscle cell calcification(71.73±2.45 ng/g vs.56.19±3.59 ng/g, P<0.01). Conclusions:Lithium chloride inhibits high phosphorus-induced vascular smooth muscle cell calcification by enhancing the level of extracellular pyrophosphate via increased ankh expression.
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Objective To evaluate the differential diagnostic value of IVIMGDWI in hepatocellular carcinoma (HCC)and atypical hepatic hemangioma (H H).Methods 3.0T MR images of 28 patients with HCC and atypical H H confirmed respectively by surgical or pathological diagnosis were retrospectively analyzed.3.0T MR routine and DWI multiple b value sequences were performed,and the related derivative parameters of IVIMGDWI (intravoxel incoherent motion diffusion,MRI)were obtained.GE ADW4.6 Functool automatic postGprocessing software and statistical methods were used for multiGparameters analyses,and the ROC curve was established to predict the value of the differential diagnosis.Results The values of ADC,slow diffusion coefficient (D)and perfusion fraction (PF)between HCC and atypical HH were statistically significant(P＝0.00,P＝0.021 ,P＜0.05 ),that was to say,there were significant differences in IVIMGDWI multiparameter comparisons between the HCC and atypical HH.And the value of fast diffusion coefficient (D?)was not statistically significant between HCC and atypical HH (P＝0.112,P＞0.05),that was to say,there was no significant difference in the multiparameter comparison between the HCC and atypical HH;The specificity and sensitivity of IVIMGDWI in HCC were from high to low:D (97.8%)＞ADC(97.4%)＞PF (82.9%),and the specificity and sensitivity of IVIMGDWI in atypical H H were D (9 7.1%)＞ADC (87.1%)＞PF (6 7.9%),from high to low,while D? was not statistically significant.Conclusion It is characteristic for the imaging manifestation of HCC and atypical HH with IVIMGDWI imaging,that is one of the important methods for clinical diagnosis and differential diagnosis of HCC and atypical HH.
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Objective To discuss the diagnostic value of direct computed tomography venography (CTV)in lower extremity deep venous thrombosis (DVT).Methods The image data of 57 lower extremity DVT cases examined by direct CTV were analyzed retrospectively,and compared with X-ray digital subtraction angiography (DSA).Results 55 cases with 63 DVT of lower extremity veins were detected by direct CTV.The positive rate of diagnosis of direct CTV was 96.49% (55/57).23 cases were peripheral type,19 cases central type, 23 cases mixed type.The imaging findings of DVT in direct CTV were the blocked or discontinued vein and filling defection,collateral circulation was showed in 8 cases.Conclusion Direct CTV may show the position,size and shape of the DVT,and is simple,safe and less contrast agent, which is one of the important method of DVT diagnosis.
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Objective@#To study the cytomorphologic features and determine whether pancreatic neuroendocrine tumors (PanNET) sampled by fine-needle aspiration (FNA) can be accurately graded based on the Ki-67 index when compared to surgical samples.@*Methods@#Corresponding intraoperative (19 cases) or endoscopic ultrasound-guided (3 cases) FNA cytology and surgical tissue specimens were obtained from 22 tumors, which were reviewed and stained for Ki-67 proliferation marker. The cytological samples included more than 200 tumor cells. Samples were graded by scoring the Ki-67 positive index in accordance with the 2010 WHO criteria. The grading scores assigned to the FNA cytology samples were compared with the scores assigned to the corresponding histological samples. Concordance was achieved by using 5% (instead of 2%) as a cut-off value for defining G2 tumors. One cytological sample included less than 500 tumor cells was excluded in the concordance calculation.@*Results@#The cytological smears consisted of uniform, monotonous and isolated cells, loose cellular aggregates and rosette-like formations. Some tumor cells clustered around segments of capillaries. The cells demonstrated distinct cytoplasmic and nuclear features. Mitoses and necrosis were rarely seen. When traditional 2% Ki-67 index cut-off value were used to classify G2 tumors, the majority (86.4%, κ=0.812, P<0.01) of FNA cytology samples and corresponding surgical tissue specimens demonstrated concordance. When a 5% cut-off value was adopted, the concordance rate was 95.5% (21/22, κ=1.000, P<0.01). Similar concordance rates between the cytological and histological grades were achieved with threshold value of cytological assessment material set at more than 500 or 200 cells.@*Conclusions@#The cytological Ki-67 index in adequate material (>200 tumor cells) is useful in grading pancreatic neuroendocrine tumors, and a cut-off value of 5% showed better predictive value compared with that of 2%. Accurate grading of PanNET is critical for predicting tumor biology, patient prognosis, and making informed decisions regarding patient management and treatment.
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The health of human, animal and environment is under serious threat from the increasing emerging infectious diseases (EID).Through strengthening the three-level disease prevention network, expanding infectious disease surveillance system and multi-sectoral joint cooperation mechanism, the quick, effective and strong prevention and control system of emerging and imported infectious diseases has been established in Shanghai.Since 2013, through effective control of EIDs such as severe acute respiratory syndrome ( SARS), influenza A H1N1 and H7N9 avian influenza, as well as imported infectious diseases (IID) such as Ebola virus disease, middle east respirators syndrome(MERS), Zika virus disease and yellow fever, the surveillance and response capacity has been improved significantly, and the prevention and control system has been improved gradually.As an international megalopolis under globalization, Shanghai is faced with the challenges as follows: growing pressure to infectious diseases prevention and control, increasingly serious situation of EIDs and IIDs;prevention and control skills need to be improved and the current personnel cannot meet with the demands.In order to meet the challenges, infectious disease monitoring and early warning technology should be strengthened; the sensitivity of infectious disease surveillance and early-warning capacity should be improved; EID symptom complex monitoring system should be established; personnel training should be strengthened, domestic and international cooperation and exchange should be carried out;so as to safeguard public health security and public health in Shanghai.
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Objective To establish cell models with different insulin sensitivity status and examine the expression of protein kinase B (PKB/Akt) in the same conditions. Methods 3T3-L1cells were cultured with different glucose concentrations (3.0, 5.5 and 50.0 mmol/L respectively).Insulin sensitivity, as presented by insulin-induced glucose transport rate in the cells, was tested by 3H-2-DG incorporation. PKB expression in cells was determined by RT-PCR. Results The insulin-induced glucose transport rate in 3T3-L1 cells was affected by different concentrations of glucose. The transport rate of 3H -2-DG in cells cultured in low glucose concentration was increased by 152% and 50% compared with high glucose concentration and control group, respectively. There was no difference in PKB mRNA expression among the three groups under the same conditions,whereas there was an obvious reduction of PKB expression in the three groups after the cells were treated with glucose and insulin. Conclusions 3T3-L1 cells cultured with different concentrations of glucose present the different insulin sensitivity status, but the difference of PKB mRNA expression has not been observed in the same condition.
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AIM To investigate the inhibitory effect of pycnogenol on generation of advanced glycation end products (AGEs) in vitro. METHODS Advanced glycation end products were determined by fluorospectrophotometer in the medium of 1 mol?L -1 glucose and 5% bovine albumin incubated at 37℃, 50℃, and 70℃ for different times. The inhibitory effect of pycnogenol was confirmed by the same system incubated with or without pycnogenol at different concentrations. RESULTS The rate of generation of AGEs in vitro was related with incubation time and incubation temperature. The generation of AGEs was inhibited by pycnogenol in vitro. The inhibitory rate was 10%～80% dependent on concentration and incubation time of pycnogenol. The inhibitory effect of pycnogenol on generation of AGEs was almost the same as that of Aminoguanidine at the same concentration. CONCLUSION pycnogenol could significantly inhibit generation of AGEs in vitro.