[Public health and clinical care integration to improve immunization of children with special health care needs].

Immunization of children with special health care needs has always been one of the difficulties in community-level public health. In recent years the relevant consensus opinions on vaccination have been issued in China, but for a long time there is a lack of effective communication channels between disease prevention and clinical medical systems in China. Pediatricians play an unoccupied role in immunization, and community-level vaccinators face the difficulties including dilemma of disease identification, evidence-based evaluation and upward referral. These lead to the vaccination hesitancy, thus the multi-disciplinary management and three-level referral model of children with special health care needs should be improved urgently. It should strengthen the integration of public health and clinical care, with the active participation of pediatricians, to promote the immunization of children with special health care needs effectively.

Research on mechanism of sevoflurane in alleviating cerebral ischemia-reperfusion injury in rats through JNK signaling pathway.

OBJECTIVE: To explore the specific mechanism of sevoflurane in alleviating cerebral ischemia-reperfusion injury (CIRI) in rats through the c-Jun N-terminal kinase (JNK) signaling pathway. MATERIALS AND METHODS: A total of 60 male specific pathogen-free Sprague-Dawley rats were randomly divided into sham group (n=20), model group (n=20), and sevoflurane group (n=20). In the sevoflurane group, sevoflurane (2.5%) was inhaled for 60 min at 24 h before the blockage of cerebral blood supply. The CIRI model was established using the suture method in the model group and sevoflurane group, while the right common carotid artery and external carotid artery were separated and ligated only, without suture placement, in the sham group. At 24 h after reperfusion, the neurological deficit score in each group was calculated, the water content in brain tissues in each group was detected based on dry-wet weight ratio, the infarction volume of brain tissues in each group was detected via 2,3,5-triphenyltetrazolium chloride (TTC) staining, and the apoptosis rate of brain cells in each group was detected using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Moreover, the protein levels of JNK, p-JNK, B-cell lymphoma-2 (Bcl-2), and the Bcl-2 associated X protein (Bax) in brain tissues were determined using Western blotting, and the gene expressions of Bax and Bcl-2 in brain tissues were determined through fluorescence quantitative Polymerase Chain Reaction (qPCR). RESULTS: It was found that the water content in brain tissues and the cerebral infarction volume were significantly increased in the model group compared with those in the sham group (p<0.01, p<0.01), while they were notably decreased in the sevoflurane group compared with those in the model group (p<0.05, p<0.01). The neurological deficit score was significantly higher in the model group than that in the sham group (p<0.01), while it was remarkably lower in the sevoflurane group than that in the model group (p<0.01). According to the results of the TUNEL assay, the model group had an evidently higher apoptosis rate of brain cells than the sham group (p<0.01), while the sevoflurane group had a lower apoptosis rate of brain cells than the model group (p<0.05). Besides, the results of Western blotting revealed that the model group exhibited remarkably increased protein levels of JNK, p-JNK, and Bax (p<0.05, p<0.01, p<0.01) and a remarkably decreased protein level of Bcl-2 (p<0.01) compared with the sham group. Sevoflurane group had decreased protein levels of JNK, p-JNK, and Bax (p<0.05, p<0.01, p<0.01) and an increased protein level of Bcl-2 (p<0.05) in comparison with the model group. In addition, the gene expression of Bcl-2 significantly declined (p<0.01), and that of Bax remarkably rose (p<0.01) in the model group compared with those in the sham group, while the contrary is the case in the sevoflurane group compared with those in the model group (p<0.05, p<0.01). CONCLUSIONS: Sevoflurane can regulate the protein and gene expressions of Bax and Bcl-2 and reduce apoptosis in CIRI by regulating the JNK signaling pathway, thereby exerting a protective effect on brain tissues and improving the symptoms of neurological deficit.

Glucagon-like peptide-1 receptor agonists and fracture risk: a network meta-analysis of randomized clinical trials.

Our network meta-analysis analyzed the effects of glucagon-like peptide-1 receptor agonists (GLP-1 RAs) on fracture risk. By combining data from randomized controlled trials, we found that GLP-1 RAs were associated with a decreased bone fracture risk, and exenatide is the best option agent with regard to the risk of fracture. This study is registered with PROSPERO (CRD42018094433). INTRODUCTION: Data on the effects of GLP-1 RAs on fracture risk are conflicted. This study aimed to analyze the available evidence on the effects of GLP-1 RAs on fracture risk in type 2 diabetes mellitus patients. METHODS: Electronic databases were searched for relevant published articles, and unpublished studies presented at ClinicalTrials.gov were searched for relevant clinical data. All analyses were performed with STATA 12.0 and R software (Version 3.4.4). We estimated the risk ratio (RR) and 95% confidence interval (CI) by combining RRs for fracture effects of included trials. RESULTS: There were 54 eligible random control trials (RCTs) with 49,602 participants, including 28,353 patients treated with GLP-1 RAs. Relative to placebo, exenatide (RR, 0.17; 95% CI 0.03-0.67) was associated with lowest risk of fracture among other GLP-1 RAs. Exenatide had the highest probability to be the safest option with regard to the risk of fracture (0.07 ‰), followed by dulaglutide (1.04%), liraglutide (1.39%), albiglutide (5.61%), lixisenatide (8.07%), and semaglutide (18.72%). A statistically significant inconsistency was observed in some comparisons. CONCLUSION: The Bayesian network meta-analysis suggests that GLP-1 RAs were associated with a decreased bone fracture risk compared to users of placebo or other anti-hyperglycemic drugs in type 2 diabetes mellitus patients, and exenatide is the best option agent with regard to the risk of fracture.

Effect of tea saponins on milk performance, milk fatty acids, and immune function in dairy cow.

This study investigated the effects of tea saponins (TSP) on milk performance, milk fatty acids, and blood immune function in dairy cows. A total of 20 early-lactation Holstein cows (days in milk = 66.4 ± 16.8 d; parity = 1.75 ± 0.91; and milk yield = 36.3 ± 7.32 kg/d; mean ± standard deviation) were randomly divided into 4 homogeneous treatment groups, with TSP added at 0, 20, 30, and 40 g/d per head, respectively. All cows had 2 wk of adaptation and 6 wk of treatments. Feed, milk, and blood were sampled and analyzed weekly. At the end of the experimental period (wk 6), the dry matter intake and yields of energy-corrected milk, milk, and milk protein, fat, and lactose in the cows fed TSP showed a quadratic response, with the lowest values in cows fed TSP at 40 g/d. The milk fat content of cows fed TSP increased linearly. Significant interactions for treatment by week were found in milk C16:1 cis-9 and C18:1 cis-9, with the highest values at wk 2, 3, and 4 in the cows fed TSP at 40 g/d. The levels declined quickly after 4 wk of feeding to values similar to those for other TSP treatments and the control at wk 5 and 6. Plasma malondialdehyde concentration decreased as the supplement level of TSP increased. The concentration of superoxide dismutase increased as the supplement level of TSP increased. The plasma concentration of tumor necrosis factor-α increased as the supplement level of TSP increased. In summary, this study showed that an intermediate dose of TSP (20 and 30 g/d) had no significant effect on feed intake, but the supplementation of 40 g/d TSP decreased feed intake, resulting in a lower milk yield. The energy-corrected milk of cows fed 40 g/d TSP declined at first but increased after 3 wk of feeding, indicating the potential adaptation to high doses of TSP supplements in dairy cows. The supplementation of TSP could reduce oxidative stress in cows and improve the immunity of dairy cows during 6 wk of feeding.

Associations between single-nucleotide polymorphisms (+45T>G, +276G>T, -11377C>G, -11391G>A) of adiponectin gene and type 2 diabetes mellitus: a systematic review and meta-analysis.

AIMS/HYPOTHESIS: The associations between adiponectin polymorphisms and type 2 diabetes have been studied widely; however, results are inconsistent. METHODS: We searched electronic literature databases and reference lists of relevant articles. A fixed or random effects model was used on the basis of heterogeneity. Sub-group and meta-regression analyses were conducted to explore the sources of heterogeneity. RESULTS: There were no statistically significant associations between +45T>G (rs2241766), +276G>T (rs1501299), -11391G>A (rs17300539) and type 2 diabetes risk. However, for -11377C>G (rs266729), the pooled OR (95% CI) for G vs C allele was 1.07 (1.03-1.11, p = 0.001). Subgroup analysis by study design revealed that -11377C>G (rs266729) dominant model (CG+GG vs CC, p = 0.0008) and G vs C allele (p = 0.0004) might be associated with type 2 diabetes risk in population-based case-control studies. After stratification by ethnicity, we found that -11377C>G (rs266729) dominant model (CG+GG vs CC, p = 0.004) and G vs C allele (p = 0.001) might be associated with type 2 diabetes risk in white individuals. In individuals with a family history of diabetes, the presence of -11391G>A (rs17300539) dominant model (GA+AA vs GG) and A vs G allele might be associated with increased risk of type 2 diabetes. CONCLUSIONS/INTERPRETATION: The presence of +45T>G (rs2241766), +276G>T (rs1501299) and -11391G>A (rs17300539) do not appear to influence the development of type 2 diabetes. However, G vs C allele of -11377C>G (rs266729) might be a risk factor for type 2 diabetes.

Vascular endothelium-specific overexpression of human catalase in cloned pigs.

The objective of this study was to develop transgenic Yucatan minipigs that overexpress human catalase (hCat) in an endothelial-specific manner. Catalase metabolizes hydrogen peroxide (H(2)O(2)), an important regulator of vascular tone that contributes to diseases such as atherosclerosis and preeclampsia. A large animal model to study reduced endothelium-derived H(2)O(2) would therefore generate valuable translational data on vascular regulation in health and disease. Yucatan minipig fetal fibroblasts stably co-transfected with human catalase (Tie2-hCat) and eGFP expression constructs were isolated into single-cell populations. The presence of the Tie2-hCat transgene in individual colonies of fibroblasts was determined by PCR. Transgenic fibroblasts were used for nuclear transfer into enucleated oocytes by electrofusion. A minimum of 140 cloned embryos were transferred per surrogate sow (n = 4). All four surrogates maintained pregnancies and piglets were delivered by cesarean section. Nine male piglets from three of the four litters carried the Tie2-hCat transgene. Expression of human catalase mRNA and overall elevated catalase protein in isolated umbilical endothelial cells from transgenic piglets were verified by RT-PCR and western blot, respectively, and endothelial localization was confirmed by immunohistochemistry. Increased enzymatic activity of catalase in transgenic versus wild-type endothelial cells was inferred based on significantly reduced levels of H(2)O(2) in culture. The similarities in swine and human cardiovascular anatomy and physiology will make this pig model a valuable source of information on the putative role of endothelium-derived H(2)O(2) in vasodilation and in the mechanisms underlying vascular health and disease.

Production of endothelial nitric oxide synthase (eNOS) over-expressing piglets.

Vascular function, vascular structure, and homeostasis are thought to be regulated in part by nitric oxide (NO) released by endothelial cell nitric oxide synthase (eNOS), and NO released by eNOS plays an important role in modulating metabolism of skeletal and cardiac muscle in health and disease. The pig is an optimal model for human diseases because of the large number of important similarities between the genomic, metabolic and cardiovascular systems of pigs and humans. To gain a better understanding of cardiovascular regulation by eNOS we produced pigs carrying an endogenous eNOS gene driven by a Tie-2 promoter and tagged with a V5 His tag. Nuclear transfer was conducted to create these animals and the effects of two different oocyte activation treatments and two different culture systems were examined. Donor cells were electrically fused to the recipient oocytes. Electrical fusion/activation (1 mM calcium in mannitol: Treatment 1) and electrical fusion (0.1 mM calcium in mannitol)/chemical activation (200 microM Thimerosal for 10 min followed by 8 mM DTT for 30 min: Treatment 2) were used. Embryos were surgically transferred to the oviducts of gilts that exhibited estrus on the day of fusion or the day of transfer. Two cloned transgenic piglets were born from Treatment 1 and low oxygen, and another two from Treatment 2 and normal oxygen. PCR, RT-PCR, Western blotting and immunohistochemistry confirmed that the pigs were transgenic, made message, made the fusion protein and that the fusion protein localized to the endothelial cells of placental vasculature from the conceptuses as did the endogenous eNOS. Thus both activation conditions and culture systems are compatible with development to term. These pigs will serve as the founders for a colony of miniature pigs that will help to elucidate the function of eNOS in regulating muscle metabolism and the cardiorespiratory system.

Oncospheres of Taenia solium develop into cysticerci in normal mice.

In vitro-hatched oncospheres of Taenia solium, prepared by the sodium hypochlorite method and adjusted to approximately 5 x 10(2)/2ml phosphate buffer saline, were injected intramuscularly or intravenously into normal Balb/c mice. When these mice were sacrificed 2 months later, all cysticerci were exclusively recovered in the lungs from the mice with intravenous inoculation, but not with intramuscular injection. A high infection rate of 76% was obtained and a total of 45 cysticerci were collected from 50 mice. Thirty-five cysticerci were mature and with normal appearance but the rest were either with abnormal appearance (4) or degenerated (6). These findings give strong evidence that T. solium oncospheres may migrate to the normal mouse lung through venous circulation and develop in this organ.

Telomeres at the chromosome X(p) might be critical in limiting the proliferative potential of human cells.

Normal human somatic cells can only divide for a limited number of times. This phenomenon has been regarded as a reflection of individual aging at the cellular level. Experimental evidences suggest that a cell's division potential is limited by the physical length of telomeres that gradually shorten through successive cell divisions. At present, it is not clear whether such a limit is imposed by the overall shortening of all telomeres or the shortening of certain critical ones. Computer simulations have suggested that among the 92 telomeres in human cells, two specific telomeres might be preferentially involved in such process. Recent experiment has shown that in a culture of male human cells, the length of the telomeres at the chromosome X(p) is reserved over the later passages during clonal proliferation. This unique feature, if can be further confirmed in other cells, implies a critical role of the telomeres at X(p) in limiting the proliferation capacity of human cells.

Influence of cholesterol on the biophysical properties of the sphingomyelin/DOPC binary system.

The influence of cholesterol on the sphingomyelin (SM)/dioleoylphosphatidylcholine (DOPC) binary system was investigated in various respects. Electron spin resonance (ESR) measurements reveal that the order parameter of 5DS (5-doxyl stearic acid) in SM/DOPC bilayers increases notably when the concentration of cholesterol is over 30 mol%. Membrane potential measurements indicate that the K+ permeability of the SM/DOPC bilayer decreases steeply at 40 mol% cholesterol concentration. Both these experiments suggest that cholesterol reduces the motion amplitude of hydrocarbon chains abruptly above 30 mol%. In contrast to the ordering effects on the hydrocarbon chains, (31)P-NMR results indicate that cholesterol slightly increases the motion of phosphate groups of the lipids. (31)P-NMR also raises the possibility of domain formation in the presence of cholesterol. Fluorescence-quenching experiments verified that solid domains appear in the binary system when cholesterol is present, and percolation threshold occurs at 50 mol% cholesterol concentration. The solid domains bear the properties of liquid ordered phase, which is the basic structure of caveolae and functional rafts. So this work provides an artificial model for the study of rafts and caveolae on biological membranes.

Effect of cholesterol on the formation of an interdigitated gel phase in lysophosphatidylcholine and phosphatidylcholine binary mixtures.

We previously reported that 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) forms an interdigitated gel phase in the presence of 1-palmitoyl-sn-glycero-3-phosphocholine (16:0LPC) at concentrations below 30 mol%. In the present investigation, fluorescent probe 1,6-diphenyl-1,3,5-hexatriene (DPH), X-ray diffraction, and differential scanning calorimetry (DSC) were used to investigate the effect of cholesterol on the phase behavior of 16:0LPC/DPPC binary mixtures. At 25 degrees C, 30 mol% 16:0LPC significantly decreases the DPH fluorescence intensity during the transition of DPPC from the L(beta') phase to the L(betaI) phase. However, the addition of cholesterol to 16:0LPC/DPPC mixtures results in a substantial increase in fluorescence intensity. The changes in DPH fluorescence intensity reflect the probe's redistribution from an orientation parallel to the acyl chain to the center of the bilayer, suggesting a bilayer structure transition from interdigitation to noninterdigitation. The normal repeat period of small angle X-ray diffraction patterns can be restored and a reflection appears at 0.42 nm with a broad shoulder around 0.41 nm in wide angle X-ray diffraction patterns when 10 mol% cholesterol is incorporated into 30 mol% 16:0LPC/DPPC vesicles, indicating that the mixtures are in the gel phase (L(beta')). Moreover, DSC results demonstrate that 10 mol% cholesterol is sufficient to significantly decrease the main enthalpy, cooperativity and lipid chain melting of 30 mol% 16:0LPC/DPPC binary mixtures, which are L(betaI), indicating that the transition of the interdigitated phase is more sensitive to cholesterol than that of the noninterdigitated phase. Our data imply that the interdigitated gel phase induced by 16:0LPC is prevented in the presence of 10 mol% cholesterol, but unlike ethanol, an increasing concentration of 16:0LPC is not able to restore the interdigitation structure of the lipid mixtures.

The structure and function of gramicidin A embedded in interdigitated bilayer.

The effects of phase transition from normal to interdigitated lipid bilayer on the function and structure of membrane proteins were studied using linear gramicidin (gramicidin A) as a model. Interdigitated bilayer structure of dipalmitoylphosphatidylglycerol (DPPG) liposomes that was induced by atropine could not be changed notably by intercalating of gramicidin. The K+ transportation of gramicidin in both normal and interdigitated bilayer was assayed by measuring the membrane potential. Results showed that gramicidin in interdigitated bilayer exhibited lower transport capability. Intrinsic fluorescence spectrum of gramicidin in interdigitated bilayer blue-shifted 2.8 nm from the spectrum in normal bilayer, which means that interdigitation provides a more hydrophobic environment for gramicidin. Circular dichroism measurement results indicated that the conformation of gramicidin in interdigitated bilayer is not the typical beta6.3 helix as in the normal bilayer. The results suggested that the interdigitated lipid bilayer might largely affect the structure and function of membrane proteins.

A drug-lipid interaction model: atropine induces interdigitated bilayer structure.

X-ray diffraction and fluorescence methods have been carried out to examine the effects of a drug, atropine, on the structure of model membranes: dipalmitoylphosphatidylglycerol(DPPG) multilamella vesicles(MLVs). A structural analysis by small angle x-ray diffraction shows that, with incorporation of atropine, the lamella repeating period of DPPG MLV is decreased from 5.89 nm to 4.52 nm. Using fluorescence probe 1,6-diphenyl-1,3,5-hexatriene(DPH) we find that the fluorescence intensity of DPH reduces largely at a narrow scope of atropine concentration. The experiments of fluorescence polarization of n-(9-anthroyloxyl)-stearic acid (nAS) and 16-(9-anthroyloxyl)-palmiticacid (16AP) discover that the normal polarization gradient almost disappears in DPPG/atropine system. By colligating the above results, we can draw a conclusion that atropine induces the transition from non-interdigitated to interdigitated stricture of DPPG vesicles in gel phase, which provides a sound model of drug-lipid interaction.