Hyperlipidemia is one of the main basic lesions of blood vessels and organ diseases. Traditional Chinese medicine (TCM) has been used in the prevention and treatment of hyperlipidemia with advantages such as little drug side effects and obvious efficacy. The establishment of hyperlipidemia diagnostic aid platform helped to improve treatment efficiency, and provide a platform for data acquisition, storage and analysis for research on hyperlipidemia syndrome standardization. At first, a collection system was implemented, which was consisted of tongue image and the text information of the other three diagnostic methods, for doctor's accurate mastering of patient's clinical manifesta-tions and improvement of therapeutic effect. Through this platform, a clustering algorithm was applied to analyze clin-ical information of four diagnostic methods among 897 hyperlipemia cases, which had been classified into 5 cate-gories. Compare disease characteristics of samples in every category with syndromes category and find out the rela-tionships between them, by which the foundation of hyperlipemia syndrome differentiation standardization will be es-tablish. It provided suggestions on TCM syndrome differentiation and diagnosis.

Background and purpose: High expression of telomerase and telomere stability are two common features in tumor cell. hTERT is a catalytic subunit of telomerase, TRF2 is extremely important to maintain the length and stability of telomerase. This study was to construct the recombinant adenovirus mediated shRNA to hTERT and TRF2, and to investigate the inhibitory effects of the vector by solo-inhibiting and connect-inhibiting in the MCF-7 cells, in order to present a new approach to the gene therapy for breast cancer. Methods: rAd-hTERT and rAd-TRF2 were constructed and the expression of hTERT mRNA and TRF2 mRNA were tested by FQ-PCR 48 hours after transfecting in MCF-7 cells. Apoptosis was observed by flow cytometry 1 to 6 days after transfection. Results: ①At 48 hours after transfection, the results of FQ-PCR showed that compared to PBS group, the expression of hTERT in rAd-hTERT group was obviously decreased and the inhibition ratio was about 86%, but TRF2 had not been obviously inhibited (P>0.05);the expression of TRF2 in rAd-TRF2 group was obviously decreased and the inhibition ratio was about 80%, but hTERT had not been obviously inhibited (P>0.05);in rAd-hTERT/rAd-TRF2 group, the inhibition ratio of hTERT and TRF2 were about 88% and 85%. Comparing rAd-hTERT/rAd-TRF2 group with rAd-hTERT group and rAd-TRF2 group, there were no significant differences of inhibition ratio between hTERT gene and TRF2 gene(P>0.05). Otherwise, comparing rAd-HK group, rAd-blank group with PBS group, there were no significant differences of inhibition ratio between hTERT gene and TRF2 gene(P>0.05). ②The result of flow cytometry showed that apoptosis was induced at the first day after transfecting in rAd-hTERT group and rAd-TRF2 group, the most obvious apoptosis was in the 3rd to 5th days,at the peak in the 5th day, and decreased in the 6th day after transfection. The apoptosis ratio of rAd-hTERT group was 46.2%, rAd-TRF2 group was 43.5%. The apoptosis ratio of rAd-hTERT/rAd-TRF2 group was 46.2% at first day, 68.5% at the second day, the most obvious apoptosis was in the 3rd to 6th days and was 77.6% in the 6th days in rAd-hTERT/rAd-TRF2 group. There were significant differences in apoptosis ratio in solo-inhibiting and connect-inhibiting(P<0.05). In addition, comparing rAd-HK group, rAd-blank group with PBS group, there were no significant differences in apoptosis ratio(P>0.05). Conclusion: ①Target sequence of RNAi which aimed at hTERT gene and TRF2 gene was designed efficiently, and the RNAi expression vectors were seen in vivo study efficiently and specifically inhibited the correspond gene expression and promoted cell apoptosis in MCF-7 cells. ②rAd-hTERT vector and rAd-TRF2 vector have no synergistical effect and antagoinstical effect on inhibiting hTERT gene and TRF2 gene mRNA expressing in MCF-7, but there was synergistical effect in terms of the induction of apoptosis. So association-RNAi-technique targeting to the genes of telomere length and stability can effectively promote tumor cell into apoptosis and inhibit breast cancer cell growth. RNAi technique of connecting correlation genes is a more effective gene therapy strategy.

Objective:To investigate the effect of resveratrol ( Res ) on endoplasmic reticulum stress induced by cerebral ischemia/reperfusion(I/R)injury in rats.Methods:The seventy-two male SD rats were divided randomly into three groups(n=20):sham operation group ( group S ) , I/R group and Res-treatment group ( group R ) .Focal cerebral I/R model was induced by electrocoagulation of left middle cerebral artery and occlusion of bilateral common carotid arteries followed by reperfusion after 30 min.The rats in Res group were treated with Res(50 mg/kg)i.p.7 d before the operation,once a day for 7 d.Neurological deficits were assessed at 24 h post-injury,followed by collecting the brain tissues.Cerebral infarct size was detected by TTC staining,and the water content of brain tissue were measured by wet-dry weight method.The expression of GRP78,p-PERK and CHOP proteins were deter-mined by immunohistochemistry and Western blot analysis.Results:Compared with sham group,the neurological deficit score and the brain water content were significantly increased(P<0.05),cerebral infarct size was enlarged(P<0.05),and the expression of GRP78, p-PERK and CHOP were up-regulated in I/R group(P<0.05).At the corresponding time,compared with I/R group,the neurological deficit score and the brain water content were markedly decreased(P<0.05),cerebral infarct size was smaller(P<0.05),the level of GRP78 was notablely increased(P<0.05),while the expression of p-PERK and CHOP were down-regulated in Res group(P<0.05). Conclusion:Resveratrol plays a protection role in ischemia-reperfusion injury, through inhibiting the endoplasmic reticulum stress in rats.

The purpose of this paper is to investigate the probable molecular mechanism in mechano-transduction of the regulation of integrins and the effects of cyclic biaxial mechanical strain on proliferation and synthetic function in the osteoblasts isolated from 3-month-old female Sprague-Dawley (SD) rats. The osteoblasts were cultured in F-12 medium contained with 10% fetal bovine serum(FBS) and grown to subconfluency in Flexercell type I dishes in a humidified incubator with 5% CO2 and 95% air at 37 degrees C. Mechanical strains were applied to the cells for periods of 30 min, 2, 4 and 8 hours every day, lasting 2 days. The amplitude of mechanical strain applied to the cells were 400, 1,000 and 4,000 mu strain respectively, at a frequency of one hertz(1 Hz). Unstrained cells were used as control. The expression of integrins alpha 2, beta 1, beta 3 on the membrane of osteoblasts and proliferation activity of osteoblasts were studied with Flow Cytometry(FCM). The content of osteocalcin, carboxyterminal propeptide of type-I procollage(PICP), total protein secreted by osteoblastes were detected with the isotope labelling method. The results showed that there are actual expressions of integrins alpha 2, beta 1, beta 3 on the membrane of osteoblasts without mechanical strain and that the expression of integrins beta 1 is highest. The mechanical strain increased the expression of integrins alpha 2, beta 1, beta 3 on the membrane of osteoblasts, but the strain-related up-regulation of expression of integrins alpha 2, beta 1, beta 3 are different in various amplitude and different duration of mechanical stains. The up-regulation of expression of integrins beta 3 is most sensitive to mechanical strain. The up-regulation of expression of integrins alpha 2, beta 1, beta 3 is higher at 4,000 mu strain than at 400, 1,000 mu strain. The mechanical strain can elevate the proliferation activity and the synthetic function of osteoblast at 400, 1,000 mu strain. However, the mechanical strain increased significant the proliferation in the osteoblasts and suppressed obviously the synthetic function in the osteoblasts. In the present study, the reaction of the osteoblasts in 3 month-old rat to the mechanical stimulation suggested that 1) expressions of integrins alpha 2, beta 1, beta 3 were increased in a amplitude of strain-dependent manner; 2) the changes of expression of integrins alpha 2, beta 1, beta 3 relate close to the changes of the proliferation and synthetic function of the osteoblasts. Low amplitude of strain can increase the proliferation and the synthetic function of the osteoblasts along with up-regulation of expression of integrins alpha 2, beta 1, beta 3; while higher amplitude of strain elevated significantly the proliferation of osteoblasts and suppressed obviously the synthetic function of the osteoblasts along with up-regulation of expression of integrins alpha 2, beta 1, beta 3. The amplitude of 4,000 mu strain is an optimal amplitude as stimulus for up-regulation of expression of integrins alpha 2, beta 1, beta 3 on the membrane of osteoblasts and increase the proliferation activity, but decrease the synthetic function of osteoblasts in the present study. Accordingly it indicates that integrins have a important role in regulation of signal transduction pathway in osteoblasts as a result of mechanical strain.
Animals ; Cell Division ; Cells, Cultured ; Female ; Integrin alpha2 ; biosynthesis ; Integrin beta1 ; biosynthesis ; Integrin beta3 ; biosynthesis ; Mechanics ; Osteoblasts ; metabolism ; physiology ; Rats ; Rats, Sprague-Dawley

Extensive monocyte recruitment is an early phenomenon associated with the development of atherosclerotic lesion. Although the molecular mechanisms are not completely understood, monocyte recruitment into these early lesions may involve changes in endothelial adhesion for monocyte, in which adhesion molecules expressed by endothelial cell play an active role. In vivo, the function of endothelial cells is not only affected by the chemical factors, but also by the mechanical factors. The purpose of this article was to investigate the induction of adhesion molecules expression by synergistic effects of Lysophosphatidylcholine (Lyso-PC) and shear stress in cultured human umbilical vein endothelial cells (HUVEC). The surface expression of ICAM-1, VCAM-1 and E-selectin on HUVEC induced by Lyso-PC(30 micrograms/ml) and shear stress(2.23, 4.20 dyne/cm2) were analyzed using flow cytometry. The results showed that: Compared with what were simultaneously exposed to shear stress and Lyso-PC, activating the cells with Lyso-PC prior to shear stress, or pre-conditioning the cells exposed shear stress prior to Lyso-PC incubation, a significantly higher expression of ICAM-1 and VCAM-1(P < 0.05) was resulted. HUVEC were exposed to shear stress and Lyso-PC at the same time or treated with each agonist alone, E-selectin expression was not significantly different from the control group. However, a sequential action of the two stimuli significantly increased E-selectin expression(P < 0.05). We concluded that: a sequential action of the shear stress and Lyso-PC induced an even greater expression of ICAM-1 and VCAM-1, thus it could be understood that the flows-hear stress in combination with endothelial activated by chemical factors may increase the ability of endothelial cells to recruit leukocytes even under the mechanical environment unfavorable for cell adhesion.
Cell Adhesion Molecules ; biosynthesis ; Cells, Cultured ; Endothelium, Vascular ; cytology ; drug effects ; metabolism ; Humans ; Lysophosphatidylcholines ; pharmacology ; Stress, Mechanical ; Umbilical Veins ; cytology

The purpose of this paper is to investigate the osteogenesis and adipogenesis in bone marrow mesenchymal stem cells (MSCs) isolated from normal rats and osteoporotic rats by ovariectomy. Osteoporotic animal model was established in 3 month-old and 6 month-old female Sprague-Dawley (SD) rats by ovariectomy. Animal experiments were divided into 4 groups: 1) control-3 group; 2) ovx-3 group; 3) control-6 group and 4) ovx-6 group. MSCs were isolated by means of the density-gradient centrifugation method from each group, respectively. Colony-forming unit-fibroblast (CFU-Fs ) number, CFU-Fs size distribution and cell density in CFU-Fs of primary passage MSCs were measured at the inverted phase contrast microscope. The cell cycle and proliferation index (PI) as well as apoptosis idex (AI) of MSCs were studied by (FCM). After osteogenic induction (OSI), calcium nodes of MSCs were marked by alizarin red staining (ARS); The expression level of alkaline phosphatase(ALP) was detected by dynamics method with substrate of phosphoric acid para-Nitro benzene and the content of osteocalcin (OCN) was detected with the isotope labelling method. After adipogenic induction (ADI), lipid droplet in MSCs were detected by oil red O staining and the mRNA level of lipoprotein lipase (LPL) was measured by RT-PCR. The results showed that CFU-Fs and PI are obviously decresed and AI are increased of MSCs in OVX-3 and OVX-6 groups (P<0.05). The secretory volume of ALP and BGP of MSCs and the content of calcium nods of MSCs are lower in OVX-3 and OVX-6 groups than that in control-3 and control-6 groups after osteogenic induction (P<0.05). The number of lipid droplet and the expression level of LPL mRNA are higher in OVX-3 and OVX-6 groups than that in control-3 and control-6 (P<0.05). The result in our study suggested that depress of osteogenesis and the up-regulation of adipogenesis of MSCs in osteoporotic rats by ovariectomy may be relate close to the pathogenesis of osteoporosis.
Adipocytes ; pathology ; Animals ; Bone Marrow Cells ; pathology ; Cell Differentiation ; physiology ; Cells, Cultured ; Female ; Mesenchymal Stromal Cells ; pathology ; Osteoblasts ; pathology ; Osteoporosis ; etiology ; pathology ; Ovariectomy ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; pathology

Objective To analyze the clinical features of patients with sporadic adult-onset neuronal intranuclear inclusion disease (NIID),and raise awareness of the disease among medical workers.Methods The clinical data of two patients with pathologically confirmed adult sporadic NIID,admitted to our hospital in February 2018 and October 2018,were collected.The clinical manifestations,head MR imaging,cutaneous pathological features,treatments and prognoses were retrospectively analyzed.Results Both patients were characterized with slow progressive dementia,accompanied with diverse clinical manifestations involving the central and autonomic nervous systems.Acute encephalopathic signs occurred in both patients.Head MR imaging showed extensive leukoencephalopathy mainly in the frontal and parietal lobes;these white matter abnormalities showed hyperintensity in T2-weighted imaging and liquid attenuated inversion recovery sequences,and equal or low signal in T1-weighted imaging.Remarkably,specific curve-like high-intensity signals along the corticomedullary junction in the bilateral frontal lobe were both observed in diffusion-weighted imaging.Patient 2 with seizures showed unilateral cerebral cortical edema on head MR imaging.In both patients,skin biopsy revealed specific eosinophilic inclusion bodies in the nucleus of some sweat gland cells,adipocytes and fibroblasts.Patient one was treated with dexamethasone intravenous drip to relieve headache and vision loss,and cognitive therapy was given.The acute encephalopathy of patient two was relieved by intravenous gamma globulin.Conclusions NIID has various clinical manifestations of central,peripheral,and autonomic nerve systems.Head MR imaging characteristic changes and skin pathological biopsy contribute to the diagnosis.Immunomodulatory therapy may be effective for acute encephalopathic symptoms in NIID.

Objective:To analyze the effect of mechanical assisted sputum drainage combined with early grade 4 rehabilitation exercise on Intensive Care Unit-acquired weakness (ICU-AW) .Methods:The convenient sampling method was used to select 98 patients who were hospitalized in ICU of Xinyang Central Hospital and underwent mechanical ventilation from June 2020 to June 2021. They were randomly divided into observation group and control group, with 49 cases in each group. During the ICU stay, the control group received ICU routine nursing, and the observation group received mechanical-assisted sputum expectoration combined with early stage 4 rehabilitation exercise on the basis of the control group. The incidence of ICU-AW, duration of mechanical ventilation, length of ICU stay, diaphragm thickness and diaphragmatic thickening fraction were compared between the two groups.Results:The occurrence time of ICU-AW in the observation group was later than that in the control group, the incidence of ICU-AW in the observation group was lower than that in the control group and mechanical ventilation time and length of ICU stay in the observation group were lower than those in the control group, and the differences were statistically significant ( P<0.05) . After intervention, skeletal muscle mass, diaphragm thickness and diaphragm thickening fraction in the observation group were all higher than those in the control group, and the differences were statistically significant ( P<0.05) . The skeletal muscle mass, diaphragm thickness and diaphragm thickening fraction of the control group after the intervention were lower than those before the intervention, and the differences were statistically significant ( P<0.05) . There was no statistically significant difference in limb skeletal muscle mass, diaphragm thickness and diaphragm thickening fraction in the observation group after intervention compared with those before intervention ( P>0.05) . Conclusions:Mechanical assisted sputum drainage combined with early grade 4 rehabilitation exercise can effectively prevent the occurrence of ICU-AW in ICU patients undergoing mechanical ventilation, shorten the duration of mechanical ventilation and ICU stay, maintain limb skeletal muscle mass and diaphragm function, which benefits patients.

Objective To analyze the etiological characteristics and the variation of pathogens spectrum in hospitalized children with a-cute respiratory tract infection(ARTI)before and after COVID-19 under"the level B of management for class B"of infectious diseases(Level B for Class B)in Shenzhen,in order to provide reference for the clinical diagnosis,treatment and prevention of ARTI.Meth-ods The ARTI cases from January 8,2022 to July 30,2022 were selected as before"Level B for Class B",and the cases from Janu-ary 8,2023 to July 30,2023 were selected as after"Level B for Class B".The pharyngeal swab samples submitted for analyzing 11 common pathogens,such as COVID-19,influenza virus(Ⅳ),respiratory syncytial virus(RSV)and mycoplasma pneumoniae(MP)in the children with ARTI admitted to Shenzhen Children's Hospital.Results SARS-CoV-2 were detected as positive in 347 cases,a-mong which 225 cases were before"Level B for Class B"including 29 cases combined with other pathogens(12.89％,29/225)and human parainfluenza viruses(HPIV)was the most common(31.03％,9/29).After"Level B for Class B",SARS-CoV-2 were detec-ted as positive in 122 cases,including 28 cases combined with other pathogens(22.95％,28/122),and RSV was the most common(28.57％,8/28).There was a statistical difference between the positive rate of SARS-CoV-2 combined with other pathogens before and after"Level B for Class B"(X2=5.834,P=0.016).After"Level B for Class B",the total pathogen detection rate(positive for at least one pathogen)was 60.82％(2 864/4 709)in the spring(January 8,2023 to April 30,2023),and influenza virus A(IVA)(22.64％,1 066/4 709),rhinovirus(HRV)(19.86％,935/4 709)and RSV(13.29％,626/4 709)were the main pathogens,and there were 301 cases(6.39％,301/4 709)of mixed infections.In the summer(May 1,2023 to July 30,2023),the total detection rate of pathogens was 70.26％(4 012/5 710),among which RSV(21.63％,1 235/5 710),MP(13.91％,794/5 710),HPIV(10.05％,574/5 710)were the main pathogens,and there were 710 cases(12.43％,710/5 710)of mixed infections,all of which were significantly higher than the same period before"Level B for Class B".The difference was statistically significant(P＜0.05).Conclusion After"Level B for Class B"for COVID-19,the detection rate of 11 common pathogens increased significantly and the pathogen spectrum of ARTI changed significantly.