Objective Assembly whole mitochondrial genome sequence of Rongshui miniature pig ( RMP ) breed and analysis the structure of mitochondrion based on the next-generation sequecing method.Comparison of phylogenetic relationship and genetic diversity among different pig breeds.Methods We collected peripheral venous blood sample from RMP and constructed two paired-end sequencing libraries.A whole-genome shotgun sequencing strategy and Illumina Genome Analyser sequencing technology were used in our study.Results The mitochondrial genome of RMP consists of 13 protein coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes and the length of pig is 16888 bp.The GC content of this pig mitochondrial genome is about 44 %.Based on phlogenetic analysis, population genetic analysis, our findings confirmed that the ancestral cluster in East Asia mainly occurred among Diannan 7#pig, Hainan wild boar, Lanyu and RMP.Conclusion RMP, a typical miniature pig breed in China, is an earlier ancestor than Lanyu pig breed.

Objective:To investigate gender measurement invariance of the Perceived Social Support Scale(PSSS)in people aged 50 years and older.Methods:A total of 1013 adults(50-96 years old)in Beijing, Hunan and Shandong were tested by using PSSS.The measurement invariance of PSSS between middle-aged and elderly males and females was analyzed.Differences in PSSS total scores and subscale scores between males and females were examined.Results:The equivalence test results of each item in the questionnaire met the requirements of the metrology(△CFI≥0.010, △TLI≥0.010, △RSMEA≤0.015), indicating that the hypotheses of morphological equivalence, weak equivalence, strong equivalence and strict equivalence of PSSS were all valid in the middle-aged and elderly population regardless of gender.In addition, middle-aged and elderly females had higher scores in family support, support from friends, support from other people and perceived social support than their male counterparts( P<0.05). Conclusions:PSSS has cross-gender equivalence in middle-aged and elderly people.Thus, differences in PSSS can reflect the perceived social support level in middle-aged and elderly people of different genders.

Objective To examine the features and clinical significance of cardiopulmonary exercise testing(CPET)in elderly patients with coronary heart disease.Methods A retrospective study was conducted at the Fifth Affiliated Hospital of Zhengzhou University from September 2015 to August 2017.Fifty-nine elderly inpatients who had undergone coronary angiography and CPET were enrolled.The mean age was 61.5 years,and there were 48 males and 11 females.Based on the results of coronary angiography,patients were divided into a coronary heart disease(CAD)group(n =40)and a non-CAD group (n =19).Meanwhile,according to the degree of coronary stenosis assessed by angiography,the CAD group was further divided into two subgroups (Group A:coronary stenosis,50 ％ -70％,n =25;Group B:coronary stenosis,＞ 70％,n =15).All results of CPET were compared between the groups.Results Patients in the CAD group were significantly associated with a lower anaerobic threshold(855.8±207.0)ml/min vs.(976.5±231.8)ml/min,a lower peak VO2 (1 371.5±388.8)ml/min vs.(1652.8±435.5)ml/min,a lower percentage of peak VO2 (18.3±4.7)ml · min-1 ·kg-1 vs.(22.0±4.4)ml · min 1 · kg-1,a lower peak O2-pulse(65.6±14.8)％ vs.(85.5±14.9)％ and a lower △VO2/△WR slope to expected value(8.0 ± 1.1)ml · min-1 · W-1 vs.(9.1 ± 1.5)ml ·min-1 · W 1,compared with those in the non-CAD group(all P＜0.05).However,there was no significant difference in other indexes between the groups.Furthermore,in the CAD group,△VO2/△WRall-Ex,△VO2/△WR＞AT and △VO2/△WRRatio to expected value were lower in Group B(7.5 ± 1.3vs.8.3±1.0,t=2.317,P=0.026),(6.2±1.9 vs.7.6±1.2,t=2.815,P=0.008)and(0.7±0.2vs.0.9±0.1,t=2.957,P =0.005).There was no difference in △VO2/△WR＜AT (P＞0.05).Conclusions Exercise tolerance is lower in elderly CAD patients than those without CAD,and △VO2/△WR is decreased during the process of exercise,especially after the AT,which may be associated with low cardiac output due to left ventricular myocardial isehemia.This finding suggests that CPET can provide evidence of ischemia in patients with coronary heart disease and is valuable for the development of diagnostic and therapeutic strategies.

Background and Objectives: Mesenchymal stromal cells (MSCs)-based treatment for degeneration of intervertebral disc (IVD) has been proposed recently. We here addressed whether MSC secreted factors can rejuvenate nucleus pulposus- derived stem/progenitor cells from degenerated disc (D-NPSCs) in vitro. Methods: and Results: We analyzed the expression of MSCs and NP cell specific surface markers, pluripotency related genes, multilineage potential and cell proliferative capacity of D-NPSCs upon incubation with the conditioned medium which was collected from the umbilical cord derived MSCs (UCMSCs). Our results indicated that the conditioned medium restore the stemness of D-NPSCs by up-regulating the expression level of CD29 and CD105, pluripotency related genes OCT4 and Nanog, and NP progenitor marker Tie2. The increased stemness was accompanied by promoted cell proliferative capacity and improved osteogenic and chondrogenic differentiation potential. Conclusions Our findings suggested that the UCMSCs derived conditioned medium might be used to rejuvenate the degenerated NP stem/progenitor cells.

Beta-amyloid (Aβ) is considered to be a central event in Alzheimer's disease (AD). However, it has been found that some population have Aβ deposition in the brain and even AD related pathology without obvious cognitive impairment, which indicates that there are factors in the body to avoid or cope with Aβ damage, and this phenomenon is called Aβ tolerance. Starting from the concepts of resilience and reserve, this paper intends to sort out the epidemiology and quantitative methods, brain characteristics and influencing factors of Aβ tolerance, in order to further deepen clinicians' understanding of Aβ tolerance in AD diagnosis and treatment, so as to provide new ideas for prevention and treatment of the disease.

Objective:Evaluate the application of Fourier transform infrared spectroscopy in the identification of homology of carbapenem-resistant Escherichia coli(CREC). Methods:A total of 26 carbapenem-resistant Escherichia coli strains were isolated from 9 provinces in China in 2018. The 900-1 200 cm -1 was selected as a spectral region for the Euclidean distance calculating and average linkage clustering between all isolates.The single nucleotide polymorphism (SNP) was analyzed by whole genome sequencing (WGS). Results:Twenty-six CREC strains were divided into 14 infrared spectros copy(IR) types by FTIR. The same IR type belonged to the same sequence type type.Compared with cluster analysis based on WGS, the consistency of FTIR cluster analysis was 92.3% (24/26).Conclusions:FTIR presented excellent performance in identification of homology of CREC.Besides, with the advantages of simple operation and rapid acquisition of results, FTIR may be a useful tool in clinical labs.

Objective:To analyze the correlation between serum insulin-like growth factor binding protein-7(IGFBP7)levels and left ventricular diastolic function and exercise capacity in elderly heart failure patients with preserved ejection fraction(HFpEF).Methods:This was a prospective study.A total of 40 elderly patients in our hospital with HFpEF were selected as the HFpEF group, and 20 patients with normal left ventricular diastolic and systolic function were selected as the control group.Serum levels of IGFBP7 were measured by enzyme linked immunosorbent assay, while N-terminal pro-brain natriuretic peptide(NT-proBNP)was measured by chemi-lumin escence assay.The levels of biomarkers were compared between the two groups.Diastolic function was assessed by transthoracic echocardiography, exercise tolerance was assessed by cardiopulmonary exercise testing(CPET).Results:The levels of IGFBP7 in HFpEF group was significantly higher than control group[median value, 59.4 ng/L(52.2-70.6 ng/L) vs.50.8 ng/L(42.0-55.2 ng/L), P<0.01]. In multivariable analysis, NT-proBNP and IGFBP7 were independently associated with HFpEF(all P<0.05). The area under receiver operating characteristic curve of IGFBP7 and NT-proBNP for detecting HFpEF were 0.778 and 0.820, respectively(all P<0.01). Higher IGFBP7 was modestly correlated with NT-proBNP, left atrial volume index, early diastole velocity(E), E/peak late diastolic velocity(E/A), early mitral inflow velocity/annular diastolic velocity(E/e′), estimated right ventricular systolic pressure, peak oxygen uptake(VO 2 peak), anaerobic threshold and minute ventilation/earbon dioxide production(VE/VCO 2)(all P<0.05). Conclusions:In elderly patients with HFpEF, IGFBP7 may be a novel biomarker of diastolic function and exercise capacity.

Objective To explore the analgesic initiation mechanism of"three-manipulations and three-acupoints"of tuina on minor chronic constriction injury(minor CCI)model rats.Methods According to the random number table method,35 SD rats were randomly divided into five groups:normal group,sham group,model group,tuina group,and tuina+MK-801 group.The model group,tuina group,and tuina+MK-801 group were subjected to ligation of the right sciatic nerve trunk to establish a minor CCI rat model.The sham group was only exposed to the right sciatic nerve without ligation,and the normal group was not subjected to any operation.The normal group was not subjected to any intervention measures.On the seventh day after modeling,the model group and the sham group underwent 9 minutes of grasping restraint,while the tuina group underwent one intervention of three-manipulations(point method,dialing method,and kneading method)and three-acupoints(right"Yinmen"(BL37),"Chengshan"(BL57),and"Yanglingquan"(GB34)acupoints)with each manipulation and acupoint intervention for 1 minute for a total of 9 minutes.The tuina+MK-801 group received intrathecal injection of MK-801 from the fifth to seventh days after modeling,with a dose of 6 μg(10 μL)per day,tuina intervention was performed 30 minutes after the last intrathecal injection,and the specific operation of tuina was the same as that of the tuina group.Before modeling,after modeling,and after intervention,each group of rats was subjected to cold sensitivity threshold(CST)and mechanical withdrawal threshold(MWT)testing.After intervention,immunohistochemistry was used to detect the positive expression of cyclic guanosine monophosphate(cGMP)in the spinal dorsal horn(SDH)at L4-6 segments;protein expressions of N-methyl-D-aspartate receptor 1(NMDAR1),neurogenic nitric oxide synthase(nNOS),soluble guanylyl cyclase β(sGCβ),and protein kinase G1(PKG1)in SDH at L4-6 segments were detected by Western blotting;mRNA expressions of NMDAR1,nNOS,sGCβ,cGMP,and PKG1 in SDH at L4-6 segments were detected by real-time PCR.Results Compared with the normal and sham groups,after modeling,CST increased and MWT decreased in the model group,tuina group and tuina+MK-801 group(P＜0.05);after intervention,the positive protein expression of cGMP was increased,the protein expressions of NMDAR1,nNOS,sGCβ,and PKG1 were increased,and mRNA expressions of NMDAR1,nNOS,sGCβ,cGMP,and PKG1 were increased in SDH at L4-6 segments in the model group(P＜0.05).Compared with the model group,after intervention,CST decreased and MWT increased in the tuina group and tuina+MK-801 group(P＜0.05);the positive protein expression of cGMP was decreased,the protein expressions of NMDAR1,nNOS,sGCβ,and PKG1 were decreased,and mRNA expressions of NMDAR1,nNOS,sGCβ,cGMP,and PKG1 were decreased in SDH at L4-6 segments in the tuina group and tuina+MK-801 group(P＜0.05).Conclusion One-time tuina intervention can effectively improve the symptoms of thermal and mechanical hyperalgesia induced by peripheral nerve injury,which may initiate analgesia through the NMDAR1/cGMP/protein kinase G signaling pathway,thereby exerting immediate analgesic effect.

Objective To explore the analgesic initiation mechanism of three-manipulation and three-acupoint tuina in model rats with minor chronic constriction injury(CCI).Methods Fifty-six SD rats were divided randomly into eight groups:normal group,sham group,model 1 group,model 2 group,tuina 1 group,tuina 2 group,tuina 1+transient receptor potential vanilloid-1(TRPV1)antagonist group,and tuina 2+transient receptor potential ankyrin 1(TRPA1)antagonist group.The model,tuina,and tuina+antagonist groups were established with minor CCI models.The tuina and tuina+antagonist groups received the three-method three-point intervention(point method,dial method,kneading method,Yinmen point,Chengshan point,Yanglingquan point)7 days after modeling.The model and sham groups were subjected to grasping restraint,and the normal group received no intervention.After the respective interventions,each group was tested for changes in mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL)to detect different types of pain.The nitric oxide(NO)content of the dorsal root ganglion(DRG)was determined by the nitrate reductase method,and changes in protein and gene expression levels of components of the TRPV1/TRPA1-NO-cGMP-protein kinase G(PKG)signaling pathway in the DRG of each group were determined by enzyme-linked immunosorbent assay,Western blot,and qPCR.Results Compared with the model group,MWT and TWL were prolonged in the tuina 1 and tuina 2 groups.Expression levels of TRPV1,TRPA1,NO,soluble guanylate cyclase-β,cGMP,and PKG1 in the DRG were significantly decreased in the tuina 1,tuina 2,tuina 1+TRPV1 antagonist,and tuina 2+TRPA1 antagonist groups.Conclusions Tuina can effectively improve the symptoms of thermal and mechanical hyperalgesia caused by peripheral nerve injury after one-time intervention.Tuina can exert immediate and continuous analgesic effects via the TRPV1/TRPA1-NO-cGMP-PKG signaling pathway.

Claudin18.2(CLDN18.2)is a tight junction protein that is overexpressed in a variety of solid tumors such as gastrointestinal cancer and oesophageal cancer.It has been identified as a promising target and a potential biomarker to diagnose tumor,evaluate efficacy,and determine patient prognosis.TST001 is a recombinant humanized CLDN18.2 antibody that selectively binds to the extracellular loop of human Claudin18.2.In this study,we constructed a solid target radionuclide zirconium-89(89Zr)labled-TST001 to detect the expression of in the human stomach cancer BGC823CLDN18.2 cell lines.The[89Zr]Zr-des-ferrioxamine(DFO)-TST001 showed high radiochemical purity(RCP,＞99％)and specific activity(24.15±1.34 GBq/μmol),and was stable in 5％human serum albumin,and phosphate buffer saline(＞85％RCP at 96 h).The EC50 values of TST001 and DFO-TST001 were as high as 0.413±0.055 and 0.361±0.058 nM(P＞0.05),respectively.The radiotracer had a significantly higher average standard uptake values in CLDN18.2-positive tumors than in CLDN18.2-negative tumors(1.11±0.02 vs.0.49±0.03,P=0.0016)2 days post injection(p.i.).BGC823CLDN18.2 mice models showed high tumor/muscle ratios 96 h p.i.with[89Zr]Zr-DFO-TST001 was much higher than those of the other imaging groups.Immunohistochemistry results showed that BGC823CLDN18.2 tumors were highly positive(+++)for CLDN18.2,while those in the BGC823 group did not express CLDN18.2(-).The results of ex vivo biodistribution studies showed that there was a higher distribution in the BGC823CLDN18.2 tumor bearing mice(2.05±0.16％ID/g)than BGC823 mice(0.69±0.02％ID/g)and blocking group(0.72±0.02％ID/g).A dosimetry estimation study showed that the effective dose of[89Zr]Zr-DFO-TST001 was 0.0705 mSv/MBq,which is within the range of acceptable doses for nuclear medicine research.Taken together,these re-sults suggest that Good Manufacturing Practices produced by this immuno-positron emission tomog-raphy probe can detect CLDN18.2-overexpressing tumors.