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The present study aims to observe the change in expression of heat shock protein 90 (HSP90) along with amyloid-β (Aβ) and phosphorylated Tau (p-Tau) protein levels in the hippocampus tissue of Alzheimer's disease (AD) transgenic animal model with age. APP/PS1 transgenic mice at age of 6-, 9- and 12-month and C57BL/6J mice of the same age were used. The cognitive abilities of these animals were evaluated using a Morris water maze. Western blot or immunohistochemistry was used to detect the expressions of HSP90 and Aβ 1-42 , as well as the phosphorylation levels of Tau protein in the hippocampus. The hsp90 mRNA levels and the morphology and number of cells in the hippocampus were detected with real-time quantitative polymerase chain reaction (qRT-PCR) and Nissl staining, respectively. The results showed that compared with C57BL/6J mice of the same age, HSP90 and hsp90 mRNA expression were decreased (P < 0.05 or P < 0.01), while Aβ 1-42 and p-Tau protein levels were increased (P < 0.05 or P < 0.01) in the hippocampal tissue of APP/PS1 transgenic mice. Meanwhile, the decrease in HSP90 and hsp90 mRNA expression (P < 0.05 or P < 0.01), the increase in Aβ 1-42 and p-Tau levels (P < 0.01 or P < 0.05) in hippocampal tissue and the reduction in behavioral ability showed a progressive development with the advancing of age in the APP/PS1 transgenic mice. In conclusion, in the hippocampal tissue of APP/PS1 mice, the decrease in HSP90 expression and the increase in Aβ 1-42 and p-Tau levels together with the decline of their cognitive ability are age-dependent.

Objectives: To study the level of serum vitamin K 2 (VitK 2 ) and its association with bone metabolism markers osteocalcin (OC), type I procollagen amino-terminal peptide (PINP), and type I collagen carboxy-terminal peptide (CTX) in children., Methods: A prospective analysis was performed on 1 732 children who underwent routine physical examination from October 2020 to October 2021. The serum levels of VitK 2 and 25-hydroxy vitamin D [25(OH)D] were measured. According to age, they were divided into four groups: <1 year, 1-3 years group, >3-6 years group, and >6-14 years. A total of 309 children with 25(OH)D≥50 nmol/L were screened out, and serum levels of OC, PINP, and CTX were measured to investigate the correlation of the serum levels of OC, PINP, and CTX with serum VitK 2 levels in different age groups., Results: The prevalence rate of serum VitK 2 deficiency was 52.31% (906/1 732). The VitK 2 deficiency group had higher prevalence rates of overweight/obesity and growth pain (≥3 years of age) than the normal VitK 2 group ( P <0.05). There were differences in the prevalence rate of serum VitK 2 deficiency ( P <0.0083) and the serum level of VitK 2 ( P <0.05) between the 1-3 years group and the >6-14 years group. The <1 year group had a higher serum level of CTX and a lower serum level of PINP than the >3-6 years group and the >6-14 years group ( P <0.05). The <1 year group had a lower serum level of OC than the >6-14 years group ( P <0.05). Serum VitK 2 level was positively correlated with OC level ( r s =0.347, P <0.01), and CTX level was negatively correlated with PINP level ( r s =-0.317, P <0.01)., Conclusions: Serum VitK 2 deficiency may be associated with overweight/obesity. Serum VitK 2 may affect the level of OC and even bone health.

Objective: To analyze the effects of bone marrow mesenchyml stem cells (BMSCs) on bone alkaline phosphatase (BALP)/C-terminal telopeptide of type-Ⅰ collagen (CTX-1) expression and mechanical dynamics in rats with osteoporotic (OP) vertebral fracture., Methods: A total of 60 female Sprague-Dawley rats were evenly divided into three groups, a control group that received sham operation (sham group), a group consisting of rats with OP vertebral fracture (OP group), and the last group consisting of OP vertebral fracture rats given BMSCs treatment (BMSCs group). Comparison of the three groups of animals was made in terms of bone dynamic change, bone quantitative broadband ultrasound attenuation (BUA) measurement, and bone mineral density (BMD). HE staining was done to examine the bone histological morphological parameters of the vertebral body. Serum CTX-1 and BALP levels were determined by ELISA., Results: Mechanical comparison showed that there were significant differences in mechanical changes of L 5 vertebra body and right femur among the three experimental groups ( P <0.05). The elastic modulus and maximum load of the OP group significantly decreased compared with those of the sham group ( P <0.05). After the intervention, the maximum load and elastic modulus of the BMSCs group were significantly higher than those of the OP group ( P <0.05). Compared with the sham group, BUA and BMD values in the OP group were significantly downregulated ( P <0.05). After intervention, BUA and BMD of the BMSCs group were significantly higher than those of the OP group and were comparable to those of the sham group ( P <0.05). Compared with the sham group, the number of trabeculae in the OP group was significantly fewer, and the distribution of trabeculae was disorderly and lacked regularity. Compared with the OP group, there were more trabeculae in the BMSCs group, and their distribution was more regular. Compared with sham group, bone histological morphological parameters of the vertebral body of rats in the OP group were significantly changed--mean trabecular plate thickness (MTPT) and trabecular bone volume (TBV) parameters were significantly decreased, while mineral apposition rate (MAR) and trabecula bone surface (TRS) parameters were significantly upregulated (all P <0.05). After the experimental intervention, bone histological morphological parameters of the vertebral body in the BMSCs group showed significant improvement compared with those of the OP group ( P <0.05). Compared with the sham group, serum BALP content in the OP group was greatly decreased, while the CTX-1 level was upregulated ( P <0.05). After the intervention, the BMSCs group had higher serum BALP content than that of the OP group and substantially lower CTX-1 content than that of the OP group ( P <0.05)., Conclusion: BMSCs can improve the mechanical changes in rats with OP vertebral fracture, and can increase the maximum load and elastic modulus of bone tissue. In addition, BMSCs can upregulate the expression of BALP in serum and downregulate the expression of CTX-1, thus helping rats with OP vertebral fracture heal early., (Copyright© by Editorial Board of Journal of Sichuan University (Medical Sciences).)

Objective: Cardiac remodeling is a common pathological manifestation of various end-stage cardiovascular diseases, which leads to myocardial diastolic and systolic dysfunction, low ejection fraction which cannot meet the needs of systemic tissue and organ metabolism, and ultimate progress into heart failure. Excessive activation of the classical renin angiotensin system (RAS), which is the angiotensin-converting enzyme-angiotensin II-type 1 angiotensin II receptor axis (ACE2-Ang II-AT1R axis), plays a key role in the pathological process of cardiac remodeling and heart failure. Angiotensin-converting enzyme 2-angiotensin (1-7)-Mas axis [ACE2-Ang (1-7)-Mas axis] is an endogenous negative regulatory pathway of classical RAS, which can reduce its harmful effects. ACE2 is a monocarboxypeptidase that can hydrolyse Ang II and produce Ang (1-7), which has cardio-protective effects. Ang (1-7), via endogenous receptor Mas, plays the role of vasodilating, anti-proliferation and anti-differentiation, anti-fibrosis, anti-thrombosis and reversing myocardial remodeling. In recent years, with increasingly growing studies on the ACE2-Ang (1-7)-Mas axis, there are more understanding about their metabolic characteristics and mechanism of action. This article describes the role of ACE2 and Ang (1-7) in cardiac remodeling and heart failure and the related mechanisms, and discusses the potential benefits by regulating ACE2 activity and Ang (1-7) levels in clinical and experimental studies, hopefully providing potential therapeutic strategies.

Objective: To explore the correlation between major inflammatory factors and septic shock in intensive care unit (ICU) patients, and to provide a basis for the diagnosis and treatment of septic shock., Methods: The patients admitted to ICU of the Third People's Hospital of Datong from March 2017 to August 2018 were selected as the research objects. According to the diagnostic criteria of septic shock, the patients were divided into septic shock group and non-septic group. The basic information and inflammatory factors levels of the two groups, including age, gender, body mass index (BMI), course of disease, acute physiology and chronic health evaluation II (APACHE II), infection site and pathogenic; and C-reactive protein (CRP), procalcitonin (PCT), neutrophil lymphocyte ratio (NLR), N-terminal pro-B-type natriuretic peptide (NT-proBNP), tumor necrosis factor-α (TNF-α), γ-interferon (IFN-γ), interleukins (IL-1β, IL-2, IL-6, IL-8) at 8 hours after diagnosis, were compared. Logistic regression was used to analyze the influencing factors of septic shock in ICU patients., Results: A total of 154 ICU patients were selected, of whom 74 had septic shock. The APACHE II score of septic shock group was significantly higher than that of non-sepsis group (23.42±3.64 vs. 15.67±2.26, P < 0.05). There was no significant difference in other baseline data between the two groups. The levels of CRP, NT-proBNP, TNF-α, IFN-γ, PCT, IL-6, IL-8 in the septic shock group were significantly higher than those in the non-septic group [CRP (mg/L): 164.3±22.6 vs. 52.3±16.2, NT-proBNP (ng/L): 426.3±288.9 vs. 167.3±80.6, TNF-α (ng/L): 193.4±39.3 vs. 88.1±20.3, IFN-γ (ng/L): 133.3±52.0 vs. 97.0±56.1, PCT (ng/L): 27.6±10.2 vs. 7.3±4.1, IL-6 (ng/L): 83.0±17.6 vs. 20.9±6.4, IL-8 (ng/L): 445.8±34.0 vs. 84.0±25.7, all P < 0.05]. It was shown by Logistic regression analysis that CRP, NT-proBNP, TNF-α, PCT, IL-6 were independent risk factors for septic shock [CRP: odds ratio (OR) = 1.662, 95% confidence interval (95%CI) = 1.132-2.567; NT-proBNP: OR = 14.688, 95%CI = 3.580-20.238; TNF-α: OR = 1.093, 95%CI = 1.043-1.343; PCT: OR = 6.378, 95%CI = 4.556-12.243; IL-6: OR = 9.641, 95%CI = 2.242-13.786; all P < 0.05]., Conclusions: The levels of inflammatory factors CRP, NT-proBNP, TNF-α, PCT and IL-6 were significantly increased, which were important factors for early diagnosis of septic shock.

Objective: To investigate the diagnostic value of N-terminal pro-brain natriuretic peptide (NT-proBNP) and troponin Ⅰ (TnI) for detecting right ventricular dysfunction (RVD) in patients with non-high-risk acute pulmonary thromboembolism (APE). Methods: A retrospective analysis was performed in 96 adult patients [44 males, 52 females, aged (61±14) years] with non-high-risk APE from January 2015 to June 2016. All patients were divided into RVD group and non-RVD group according to whether there was right ventricular enlargement on echocardiography. The baseline data, serumTnI and NT-proBNP levels were compared between the 2 groups and the diagnostic value of the 2 cardiac markers for RVD was analyzed. Results: There were no significant differences in age, gender, body mass index between the 2 groups ( P> 0.05). The creatinine clearance rate of the RVD group was lower than that of the non-RVD group [96.4 (77.5,99.6) vs 101.7 (95.1,106.5), P= 0.021]. NT-proBNP [2 300 (1 056,3 396) vs 188 (61,535), P< 0.01] and TnI [0.13 (0.09,0.25) vs 0.00 (0.00,0.02), P< 0.01] were significant higher in the RVD group than in the non-RVD group. The univariate logistic regression analyses showed that NT-proBNP (per 100 ng/L, OR 1.199, 95% CI 1.117-1.287), TnI (per 0.01 μg/L, OR 1.164, 95% CI 1.079-1.256) and creatinine clearance rate ( OR 0.968, 95% CI 0.938-0.998) were significantly associated with RVD. Multivariate regression analysis showed that NT-proBNP (per 100 ng/L, OR 1.155, 95% CI 1.074-1.241) and TnI (per 0.01 μg/L, OR 1.079, 95% CI 1.011-1.151) were independently associated with RVD. The areas under the ROC curve (AUC) of NT-proBNP, TnI, and the combination of them were 0.908 (95% CI 0.841-0.976), 0.896 (95% CI 0.826-0.966) and 0.925 (95% CI 0.862-0.988), respectively. The cut-off value of NT-proBNP was 503.5 ng/L, with a sensitivity of 85.7%, specificity of 75.4%, positive predictive value (PPV) of 66.7% and negative predictive value (NPV) of 90.2%.The cut-off value of TnI was 0.05 μg/L, and the sensitivity, specificity, PPV and NPV was 80.0%, 86.9%, 77.8% and 88.3%, respectively. The optimal probability derived from the logistic regression model in which the 2 biomarkers were the independent variables was 0.779, with a sensitivity, specificity, PPV and NPV of 65.7%, 96.7%, 92.0%, 83.1%, respectively. Conclusion: Both NT-proBNP and TnI had preferably good diagnostic value for RVD in patients with non-high-risk APE, but their clinical application needed comprehensive evaluation combined with the overall manifestations of the patients and experimental methods. The diagnostic value was higher when the 2 biomarkers were evaluated together.

Epithelial-mesenchymal transition (EMT) occurring in alveolar epithelial cells plays an important role in the development and progression of pulmonary fibrosis. Previous studies showed that antiflammin-1 (the active fragment of uteroglobin) effectively inhibited bleomycin-induced pulmonary fibrosis. However, its mechanism is still far from being clarified. In this study, we investigated the effects of antiflammin-1 on EMT in A549 cells induced by transforming growth factor-β1 (TGF-β1) and the underlying mechanism by using morphological observation and Western blot. The results showed that the expression of α-smooth muscle actin (α-SMA) increased significantly while the expression of E-cadherin decreased significantly in A549 cells following treatment with TGF-β1 concomitant with morphological change of A549 cells from pebble-like shape epithelial cells to spindle-like mesenchymal shape. This process of EMT in A549 cells induced by TGF-β1 was significantly inhibited when A549 cells were co-incubated with TGF-β1 and antiflammin-1. Furthermore, the anti-lipocalin interacting membrane receptor (LIMR) antibody and PD98059 (an ERK signaling pathway blocker) attenuated the inhibitory effect of antiflammin-1 on TGF-β1-induced EMT, respectively. Our findings indicate that antiflammin-1 can inhibit EMT in A549 cells induced by TGF-β1, which is related to LIMR and its downstream ERK signaling pathway.

Objective To investigate the role of aliskiren (AL) in the angiotensinII/ angiotensin 1-7 (AngII/Ang1-7) signal pathway in renal tissue of diabetic nephropathy (DN) rats. Methods 40 male SD rats were randomly divided into 4 groups including normal group, diabetes group, AL group and valsartan (Val) group. Animal models of diabetes were induced by high fat diet combined with small dose of streptozotocin injection. Rats in AL group were administered 50 mg/(kg.d) AL by gavage, and rats in Val group were administered 30 mg/(kg.d)Val by gavage. 24-hour urine protein (24 h-UP) were observed by Coomassie blue colorimetry at 2, 4 and 6 weeks after modeling, serum creatinine was detected by automatic biochemical analyser, kidney index [kidneys mass(g)/body mass (kg)] was measured. HE and PAS staining were used to observe renal pathological changes. Immunohistochemistry was used to detect the expression of ACE2, MAS receptor, AT1R and AT2R in the kidney. Results After 6 weeks of modeling, there was no significant difference in creatinine level among groups. The levels of glucose, 24 h-UP and kidney index in diabetes group, AL group and Val group significantly increased. Compared with diabetes group, the levels of 24 h-UP and kidney index were lower in AL group and Val group. Immunohistochemistry showed that the expression of AT2R and ACE2 was lower and the expression of MAS receptor was higher in AL group than diabetes group and Val group. Compared to normal group and Val group, AT1R expression was significantly up-regulated in AL group, without significant difference between diabetes group and AL group. Conclusion AL down-regulates the expression of AT2R and ACE2, thus inhibits the AngII/ Ang1-7 signal axis and improves/alleviates the symptoms in diabetic rats.

Objective: To investigate the effect of caveolin-1 scaffolding domain (CSD) peptides on heme oxygenase-1 (HO-1) activity increasing and M1/M2 phenotype polarization in rat alveolar macrophages (AMs) induced by lipopolysaccharide (LPS)., Methods: Bioinformatics was used to analyze the binding of full-length wild-type CSD polypeptide and 101 amino acid deleted truncated mutant CSD polypeptide (Δ101CSD) to HO-1. Primary AMs were isolated from rats, when cell fusion reached 80%, they were synchronized with serum-free medium and divided into five groups: no treatment was given to the blank control group; LPS group was treated with 100 μg/L LPS for 16 hours; LPS + hemin group was treated with 100 μg/L LPS and 20 μmol/L hemin for 16 hours; wild-type CSD polypeptide + LPS + hemin group was pretreated with 10 μmol/L wild-type CSD polypeptide 6 hours before LPS treatment; Δ101CSD + LPS + hemin group was pretreated with 10 μmol/L Δ101CSD polypeptide 6 hours before LPS treatment. After treatment for 16 hours, the co-localization between caveolin-1 (Cav-1) and HO-1 was displayed by confocal microscope; the mRNA expressions of inflammatory cytokines interleukin-1β (IL-1β) and monocyte chemoattractant protein-1 (MCP-1) and M1/M2 polarization cytokines tumor necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS), leukocyte differentiation antigen 206 (CD206) and IL-10 were determined by real-time fluorescent quantitative reverse transcription-polymerase chain reaction (RT-qPCR); the HO-1 activity and nitric oxide (NO) production were determined by spectrophotometry., Results: Bioinformatics analysis showed: both wild-type CSD and Δ101CSD peptides could bind to HO-1, and there was no significant difference in the binding ability between the two peptides, but the deletion of 101 Arg resulted in the disappearance of part of the binding region between Δ101CSD and HO-1. The results of laser confocal microscopy showed: the expressions of Cav-1 and HO-1 were lowed in the blank control group, and Cav-1 was bound to HO-1 in LPS group and LPS + hemin group. Both wild-type CSD and Δ101CSD peptides pretreatment could significantly reduce the binding of HO-1 to Cav-1 induced by LPS. HO-1 activity analysis showed: after LPS stimulation, the activity of HO-1 was significantly higher than that of the blank control group; the activity of HO-1 induced by LPS was increased by hemin; after pretreatment with two kinds of CSD peptides, the activity of HO-1 was further increased, and the effect of wild-type CSD peptide was more significant, which showed a statistically significant difference as compared with that of LPS + hemin group (pmol×mg -1 ×h -1 : 3 683±266 vs. 2 408±132, P < 0.05). RT-qPCR results showed: LPS could induce elevation of cytokines and M1 markers and decrease of M2 markers, while hemin could inhibit LPS-induced inflammatory response and M1/M2 phenotypic polarization. Compared with LPS + hemin group, after pretreatment with wild-type CSD peptide, the levels of inflammatory factors in AMs were decreased, and the mRNA expression levels of TNF-α and iNOS, M1 markers, were decreased [TNF-α mRNA (2 -ΔΔCt ): 6.82±0.05 vs. 8.70±0.24, iNOS mRNA (2 -ΔΔCt ): 331.50±32.05 vs. 506.70±0.10, both P < 0.05], and IL-10 mRNA expression level was increased (2 -ΔΔCt : 269.09±6.54 vs. 119.05±3.30, P < 0.05). The deletion of 101 site partially weakened the inhibitory effect of CSD peptides on inflammatory factors and only reduced the expression of iNOS mRNA (2 -ΔΔCt : 429.11±8.92 vs. 506.70±0.10, P < 0.05), indicating that its ability to transform AMs from M1 phenotype to M2 phenotype was poor. The two peptides had no effect on the expression of CD206., Conclusions: Wild-type CSD had beneficial effects of anti-inflammation by reducing Cav-1 binding to HO-1 induced by LPS, restoring the HO-1 activity and driving M2 phenotype in alveolar macrophages.

This study was aimed to explore the effect of angiotensin converting enzyme 2 (ACE2) agonist diminazene aceturate (DIZE) on acute lung injury (ALI) induced by limb ischemia-reperfusion (LIR) in mice. Male 8-week-old wild-type and hACE2 transgenic ICR mice were randomly divided into 6 groups (6 in each group), including wild-type control (W), wild-type model (WL), wild-type model with DIZE administration (WLD), transgenic control (T), transgenic model (TL), and transgenic model with DIZE administration (TLD) groups. LIR model was established by 4 h reperfusion following 2 h ischemia of bilateral hindlimbs with rubber bands in mice. The WLD and TLD groups were pretreated with DIZE (15 mg/kg, i.p.) for 4 weeks before LIR. At the end of LIR, the mice were sacrificed and lung tissues were sampled. Indexes for evaluating lung injury include organ coefficient and wet/dry weight ratio (W/D), cell count and protein concentration of bronchoalveolar lavage fluid (BALF), as well as morphological change and pathological score were detected. Angiotensin II (Ang II) and Ang (1-7) levels in lung tissue were determined by using ELISA commercial kits. And the protein expressions of angiotensin II type 1 receptor (AT1) and Mas receptor protein in lung tissue were detected by Western blot. The results were as follows: (1) There was obvious lung injury in both the WL and TL groups. The lung injury in the TL group was lighter than that in the WL group. DIZE could attenuate the lung injury in both the two groups. (2) The WL group showed increased Ang II and decreased Ang (1-7) levels, whereas the TL group did not exhibit any changes of these two proteins. DIZE decreased the level of Ang II in both the WL and TL groups, and increased the level of Ang (1-7) in the WL group. (3) In the WL and TL groups, AT1 and Mas receptor protein expressions were up-regulated. DIZE reversed the change of AT1 protein expression, whereas further increased Mas receptor expression in both the two groups. These results suggest that DIZE may improve the renin-angiotensin system homeostasis by regulating ACE2-Ang (1-7)-Mas axis in local lung tissue and play a protective role in LIR-induced ALI in mice.

Objectives: To study the expression pattern of brain natriuretic peptide （BNP） in myocardial tissue from forensic routine cases and to explore its application value in the forensic determination of sudden cardiac death （SCD）., Methods: The data of 96 autopsy cases accepted by the center of Medico-legal Investigation of China Medical University between December 2008 to May 2014 were collected. There were 62 cases in SCD group cardiac and 34 cases in non-SCD group. The myocardial tissues were taken from left and right ventricular wall, respectively. The expressions of BNP protein and BNP mRNA in myocardial tissue were detected by HE staining, immunohistochemical staining, Western blotting and quantitative real-time reverse transcription-polymerase chain reaction （RT-qPCR）, etc., Results: The immunohistochemical staining of myocardial tissue showed diffusely positive staining in SCD group, and patchily or diffusely positive staining in non-SCD group with lighter degree. The result of Western blotting showed that the expression of BNP protein elevated in left ventricular wall of SCD group. The result of RT-qPCR showed a positive correlation between the BNP mRNA expressions in bilateral ventricular walls and the heart weight, bilateral lung weight, and N-terminal prohormone of brain natriuretic peptide （NT-proBNP） concentration. There were large differences between the BNP mRNA concentrations in SCD group and non-SCD group, and the former was statistically higher （ P <0.05）., Conclusions: The expressions of BNP protein and BNP mRNA in myocardial tissue are related to the causes of death. Combined with pathological changes, the expressions of BNP protein and BNP mRNA in myocardial tissue have certainly practical significance for the determination of SCD and the analysis of the death mechanism in the cases related to forensic pathology., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Editorial Department of Journal of Forensic Medicine)

Objectives: To study the expression change of pro-brain natriuretic peptide （proBNP） and N-terminal pro-brain natriuretic peptide （NT-proBNP） in sudden death of coronary atherosclerotic heart disease, and to explore its application in forensic diagnosis., Methods: Myocardial and blood samples were collected from normal control group, sudden death of coronary atherosclerotic heart disease group and single coronary stenosis group （20 cases in each group）. The expression of proBNP in myocardial samples were detected by immunohistochemical staining and Western blotting, and that of BNP mRNA were detected by reverse transcription PCR （RT-PCR）. The content of NT-proBNP in plasma were detected by ELISA., Results: Immunohistochemical staining showed positive expression of proBNP in both sudden death of coronary atherosclerotic heart disease group and single coronary stenosis group. There was no positive expression in normal control group. For sudden death of coronary atherosclerotic heart disease group and single coronary stenosis group, the relative expression of proBNP protein and BNP mRNA in myocardial tissue and the NT-proBNP content in plasma were higher than that of normal control group （ P <0.05）. The NT-proBNP content in plasma of sudden death of coronary atherosclerotic heart disease group was higher than that of single coronary stenosis group （ P <0.05）., Conclusions: In myocardial ischemia condition, the higher expression of proBNP in cardiac muscle cell shows that the detection of NT-proBNP in plasma can be useful to differentially diagnose the degree of coronary atherosclerotic heart disease and determine whether the sudden death due to coronary atherosclerotic heart disease., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Editorial Department of Journal of Forensic Medicine)

Background: The aim of this study is to explore roles of thyroid transcription factor-1 (TTF-1), CD56, P40 expression and other clinical characteristics predicting response and survival in patients with small cell lung cancer (SCLC)., Methods: Formalin-fixed, paraffin-embedded biopsy tissues were retrospectively obtained from 198 SCLC patients who were diagnosed first in Xuanwu Hospital. The expressions of TTF-1, CD56 and P40 were detected by immunohistochemistry. The clinical data including age, gender, cancer stage, Eastern Cooperative Oncology Group (ECOG) score, smoking or not, superior vena cava syndrome (SVCS) due to lung cancer or not were collected. Cox proportional hazard model was used to analyze the relationship between the overall survival (OS) and factors., Results: Immunohistochemical staining results showed the positive rate of TTF-1, CD56, P40 were 73.2%, 88.4% and 7.1% respectively. TTF-1 expression (OR=0.665, 95%CI: 0.472-0.937), smoking index ≤400 (OR=1.72, 95%CI: 1.061-2.789) and ECOG=2 (OR=3.551, 95%CI: 2.133-5.914), extensive-stage (OR=2.487, 95%CI: 1.793-3.451) and SVCS due to lung cancer (OR=2.394, 95%CI: 1.49-3.846) were independent prognostic factors for SCLC patients., Conclusions: Prognosis of SCLC was related to TTF-1 expression independently after adjusting smoking, ECOG score, stage and SVCS due to lung cancer. Detection of TTF-1, CD56 and P40 expression level might be helpful for predict the prognosis of SCLC.
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Objective: To observe the influence of electroacupuncture (EA) stimulation with different electrical current intensities and therapeutic intervals on learning-memory ability and expression of β-amyloid peptide Aβ 1-40 and arginine vasopressin (AVP) genes in the hippocampal CA 1 region in vascular dementia (VD) rats, so as to provide evidence for treatment of VD., Methods: A total of 48 male SD rats were randomly divided into sham, model, 0.5 mA-5 d-EA, 1.5 mA-5 d-EA, 0.5 mA-1 d-EA and 1.5 mA-1 d-EA groups ( n =8 in each group). The VD model was established using modified 4-vessels occlusion method. EA (alternative 2 Hz/15 Hz, 0.5 mA, 1.5 mA) was applied to "Baihui" (GV 20) and "Dazhui" (GV 14) for 30 min, once a day (1 d) or once every 5 d for 10 times. The learning-memory ability was detected using Morris water maze tests (place navigation task and spatial probe trials), and the expression of Aβ 1-40 and AVP genes in hippocampal CA 1 region was determined using real time-PCR., Results: In comparison with the sham group, the average escape latency of place navigation task, and the duration for crossing the target-platform for the 1 st time (spatial navigation task) were significantly increased ( P <0.05), and the times to cross the target-platform within 2 min (spatial probe trials) were significantly decreased in the model group ( P <0.05), suggesting a reduction of learning-memory ability. The expression level of Aβ 1-40 mRNA was remarkably increased ( P <0.05), and that of AVP mRNA notably decreased in the model group ( P <0.05). Following EA intervention, the increased escape latency and the duration of crossing the target-platform for the 1 st time and the increased Aβ 1-40 mRNA expression, and the decreased target-platform crossing times within 2 min as well the down-regulated AVP mRNA expression level were reversed in the 4 EA groups compared with the model group ( P <0.05). The therapeutic effects of higher stimulating intensity (1.5 mA-1 d-EA and 1.5 mA-5 d-EA) were markedly superior to those of lower intensity (0.5 mA-1 d-EA and 0.5 mA-5 d-EA), and the effects of higher frequency of EA intervention (0.5 mA-5 d-EA and 1.5 mA-5 d-EA) were obviously superior to those of lower frequency of EA (0.5 mA-1 d-EA and 1.5 mA-1 d-EA) in suppressing the increased 3 indexes and the decreased 2 indexes mentioned above ( P <0.05)., Conclusions: EA can improve the learning-memory ability of VD rats, which Feb be related to its effects in inhibiting the expression of Aβ 1-40 mRNA and up-regulating the expression of AVP mRNA in hippocampal CA 1 region; and the therapeutic effects of higher stimulating intensity and higher intervention frequency are obviously better than those of lower stimulating intensity and lower therapeutic frequency.

Objective: To explore the difference in end tidal PCO(2) (P(ET)CO(2)) between idiopathic pulmonary arterial hypertension (IPAH) and chronic thromboembolic pulmonary hypertension (CTEPH), and to analyze the correlation between P(ET)CO(2) and the indexes of disease severity in IPAH and CTEPH patients. Methods: Data were retrieved from 68 IPAH patients and 52 CTEPH patients who all had received right-heart catheterization, pulmonary function test and cardiopulmonary exercise testing at Shanghai Pulmonary Hospital from October 2011 to October 2014. In addition, other clinical parameters were also collected. Results: The IPAH group had a significantly higher mPAP, PVR [(60±16) mmHg (1 mmHg=0.133 kPa), (13±6) Wood U ] than the CTEPH group [(46±12) mmHg, (9±4) Wood U, t =4.90, 4.83, all P <0.01]. Meanwhile, the IPAH group had a lower percentage of predicted peakVO(2), oxygen pulse [(45±15)%, (60±22)%] compared with the CTEPH group [(53±16)%, (68±21)%, t =-2.42, -1.96, all P <0.05]. The value of P(ET)CO(2) at rest, AT, peak in the IPAH patients [(27±5), (28±7), (25±7) mmHg] were higher than those in the CTEPH patients [(24±4) mmHg, (23±6) mmHg, (21±6) mmHg, t =3.22-4.54, all P <0.01]. There was a significantly difference in P(ET)CO(2) at AT and peak between WHO-FC Ⅰ-Ⅱ and Ⅲ-Ⅳ subgroups in IPAH ( t =2.55, 2.60, all P <0.05) and CTEPH ( t =2.39, P <0.05), except for P(ET)CO(2) at peak in the CTEPH patients ( t =1.71, P >0.05). A moderately inverse correlation was found between P(ET)CO(2) at AT and NT-proBNP in the IPAH group ( r =-0.58, P <0.01), meanwhile P(ET)CO(2) at AT in the CTEPH group was weakly correlated with NT-proBNP ( r =-0.34, P <0.05). Conclusions: Compared with the CTEPH patients, the IPAH patients had significantly decreased exercise capacity and increased P(ET)CO(2). P(ET)CO(2) could reflect the disease severity in both IPAH and CTEPH patients, being superior in IPAH than in CTEPH. Furthermore, P(ET)CO(2) at AT might be better than P(ET)CO(2) at peak in reflecting the ventilatory efficiency.

Objective: To observe the change of angiotensin converting enzyme (ACE) and ACE2 in the murine myocardium followed hemorrhagic shock and the role of post-hemorrhagic shock mesenteric lymph (PHSML) drainage., Methods: Twenty-four male mice were ran-domly divided into control, sham, shock, and shock + drainage groups. A hemorrhagic shock model was established and then fluid resuscita-tion was performed to the mice in the shock and shock + drainage groups, and the PHMSL was drained in the shock + drainage group after fluid resuscitation. After 6 h of resuscitation in the shock and shock + drainage groups or corresponding time in the sham group, or after anesthesia in the control group, the myocardial tissues were harvested for the determination of the mRNA expressions of ACE, ACE2, angiotensin Ⅱ (Ang Ⅱ) type 1 receptor (AT1R), and Mas related G protein coupled receptor (Mas1R) using the method of qRT-PCR, and the levels of Ang Ⅱ and Ang (1-7) using the method of ELISA., Results: In the myocardial tissue of shock group, the ACE and AT1R mRNA expressions and Ang Ⅱ level were significantly increased than those of the control and sham groups, the ACE2 and Mas1R mRNA expressions were significantly de-creased than that of the control and sham groups, the Ang (1-7) level was decreased compared with the control group, the ratios of ACE/ACE2, Ang Ⅱ/Ang (1-7), and AT1R/Mas1R in the shock group were significantly increased than the control and sham groups. Meanwhile, PHSML drainage obviously suppressed the effects of hemorrhagic shock on these indices., Conclusions: Hemorrhagic shock up-regulated the ACE-Ang Ⅱ-AT1R axis, down-regulated the ACE2-Ang (1-7)-Mas1R axis, and induced the unbalance of ACE and ACE2 in myocardial tis-sue. PHSML drainage decreased the ACE-Ang Ⅱ-AT1R axis and increased the ACE2-Ang (1-7)-Mas1R axis, resulted in the balance of ACE and ACE2.

Objective: To investigate the role of lymph vessel density (LVD) and microvessel pericyte coverage index (MPI) in the pathogenesis of nasal polyps. Methods: Using immunohistochemistry and immunofluorescence double staining method, the expressions of albumin, D2-40 and CD34-α-SMA in 11 specimens of normal nasal mucosa, 26 specimens of nasal polyp and 26 specimens of inferior turbinate tissue from patients with nasal polyps were investigated. The counts of microvessel density (MVD), lymph vessel density (LVD) and microvessel pericyte coverage index (MPI) were compared. SPSS 17.0 software was used to analyze the data. Results: The nasal polyp group(0.269±0.096) had more albumin than inferior turbinate tissue group(0.159±0.078) and normal nasal mucosa group(0.138±0.045), the differences were significant ( q value was 4.873, 4.446, both P <0.05). The counts of MVD in nasal polyp group (30.52±4.42) were not significantly higher than those in inferior turbinate tissue group (30.33±6.03) and normal nasal mucosa group(28.21±6.84), the differences were not significant ( q value was 0.130, 1.147, both P >0.05). The MPI in nasal polyp group (0.291±0.096) was significantly lower than those in inferior turbinate tissue group(0.432±0.101) and normal nasal mucosa group(0.416±0.071), the difference was significant ( q value was 5.399, 3.680, both P <0.05). The counts of LVD in the nasal polyp group(0.245±0.073) were significantly lower than those in inferior turbinate tissue group (0.431±0.054) and normal nasal mucosa group(0.470±0.078), the difference was significant ( q value was 10.004, 9.328, both P <0.05). MPI expression in the nasal polyp group was negetively correlated to albumin expression( r =-0.889, P <0.05). The LVD expression in the nasal polyp group was negetively correlated to albumin expression( r =-0.901, P <0.05). Conclusion: Different LVD and MIP in nasal polyp tissues and normal nasal mucosa tissues imply that microcirculatory dysfunction plays a crucial role in the pathogenesis of nasal polyps.

To investigate the effect of the hyperforin (HF) on learning and memory function and Aβ₁₋₄₂, βAPP and BACE1 protein expressions in hippocampus of five-month-old APP/PS1 double transgenic mice, and discuss the underlying mechanism of HF. The five-month-old APP/PS1 double transgenic mice were randomly divided into the model group, rosiglitazone group (12 mg•kg⁻¹•d⁻¹) and HF high dose, middle dose and low dose groups (600, 300 and 150 mg•kg⁻¹•d⁻¹) in each group; in addition, 15C57BL/6J mice with the same months and background were selected as normal group. Drugs were diluted in the same volume before using, and then administrated by ig for 7 months, 1 time a day; the mice in normal group and model group received the same volume of distilled water. The learning and memory ability was tested by Morris water maze; Aβ₁₋₄₂, βAPP and BACE1proteinexpressionlevelswere tested by immunohistochemistry and Western blot. The Morris water maze results showed that as compared with the normal group, the learning and memory ability was significantly impaired in mice of model group (P<0.01); as compared with the model group, the learning and memory ability was improved in mice of rosiglitazone group and HF high, middle and low dose groups(P<0.01 or P<0.05). Immunohistochemistry and western blot results showed thatas compared with the normal group, the Aβ₁₋₄₂, βAPP and BACE1 protein expression levels in hippocampus were significantly increased in mice of model group (P<0.01);as compared with the model group, Aβ₁₋₄₂, βAPP and BACE1 protein expression levels in hippocampus were decreased in mice of rosiglitazone group and HF high, middle and low dose groups (P<0.01 or P<0.05). HF may improve the learning and memory ability of AD model mice via inhibition of βAPP and BACE1 protein expressions, thus reduced the generation of Aβ₁₋₄₂ proteins and amyloid plaque deposits in the brain., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

Objective: To observe analgesic effect of electroacupuncture ( EA) on rats with chronic inflammatory pain and its regulatory mechanism on ispilateral dorsal root ganglion (DRG) and spinal dorsal horn (SDH) Mas-related G protein-coupled C receptor (MrgprC)., Methods: Totally 40 healthy male SD rats were divided into 4 groups according to random number table, i.e., the normal (N) group, the model (M) group, the acupuncture (Acu) group, the EA group, 10 rats in each group. The model of chronic inflammatory pain was established by subcutaneous injecting 0. 1 mL complete Freund's adjuvant (CFA) into right hind paw. Paw withdrawal thresholds (PWTs) were measured before modeling, at day 1, 3, 5, 7, and after CFA injection, respectively. Expression levels of MrgprC in ispilateral DRG and SDH were detected by Western blot. The content of bovine adrenal medulla 22 (BAM22) in SDH was detected by immunohistochemical assay., Results: Compared with N group at each time point, PWTs significantly decreased in M group (P <0. 01). Compared with M group, PWTs significantly increased at day 5 of EA and after EA in EA group (P < 0.05, P < 0.01). Compared with Acu group at each time point, post-EA PWTs significantly increased in the EA group (P < 0.05). Compared with N group, expression of MrgprC in ispilateral DRG and ratio of BAM22 positive cells in ispilateral SDH increased in M group (P < 0.01). Compared with M group, expression of MrgprC in ispilateral DRG and ratio of BAM22 positive cells in ispilateral SDH increased in the EA group (P < 0.05)., Conclusion: EA had favorable analgesic effect on chronic inflammatory pain induced by CFA, and its mechanism might be possibly associated with up-regulating MrgprC expression in ispilateral DRG and BAM22 content in ispilateral SDH.

Objective: To observe the clinical characteristics of bone disease in patients with multiple myeloma (MM) and the clinical significance of monitoring bone metabolic markers., Methods: The data of 178 MM cases newly diagnosed in Beijing Ji Shui Tan Hospital from January 2009 to June 2014 were reviewed to analysis the types and classification of bone disease and to observe the clinical characteristics of patients with different grades of bone disease. The levels of bone metabolic markers total procollagen type Ⅰ N-terminal peptide (tPINP) and β C-terminal telopeptide of type Ⅰ collagen (β-CTX) were monitored regularly in the two years following treatment in 66 cases., Results: (1) Among the 178 newly diagnosed MM cases, 167 cases complained of pain in bones on first visit, 35 cases combined with hypercalcemia, 83 cases combined with osteoporosis, 154 cases combined with osteolytic bone destruction, and 73 cases combined with pathologic fracture. The most common osteolytic location was the spine. The most common fracture sites was the spine. (2) According to bone disease grading, the 178 cases were divided into group A (bone grade 0-2, n=51) and group B(bone grade 3-4, n=127). There were no significant differences between group A and group B in gender, median age, therapeutic effect/ineffec, median overall survival, median progress-free survival, mean serum lactic dehydrogenase, mean albumin, urine light chains and serum creatinine(all P>0.05). Compared with group A, group B had lower hemoglobin level[(99.78±29.93)vs (108.84±29.30) g/L], and higher blood calcium level[(2.47±0.40)vs (2.30±0.29) mmol/L], serum β2-microglobuin level[(6.04±4.84)vs (4.12±3.97)mg/L], and bone marrow plasma cells percentage(33.30%±24.87% vs 23.51%±22.67%)(all P<0.05). (3) Before treatment, the levels of β-CTX and tPINP in patients of group B(n=47) were higher than those in group A(n=19)(median 0.78 vs 0.42 μg/L, 60.95 vs 43.47 μg/L, both P<0.05). The ratio of β-CTX /tPINP in group B was higher than that in group A (median 0.017 vs 0.012, P<0.05). After chemotherapy for 3 months, there were no differences in the level of tPINP compared with that before treatment in both group A and group B (both P>0.05), the level of β-CTX decreased significantly compared with that before treatment in both groups(median 0.16 vs 0.42 μg/L, 0.26 vs 0.78 μg/L, both P<0.05); the ratio of β-CTX /tPINP decreased significantly compared with that before treatment in both group A and in group B(median 0.008 vs 0.012, 0.011 vs 0.017, both P<0.05). There were no differences in the level of β-CTX, tPINP and β-CTX/tPINP ratio after treatment for 6 months, 1 year and 2 years compared with that after 3 months in both group A and group B (all P>0.05). (4)All patients were divided into two groups according to the therapeutic effect: effective group included patients who reach the effect of partial remission or better remission(n=48), while ineffective group included patients who did not reach the effect of partial remission(n=18). Before treatment there were no differences in the level of β-CTX, tPINP and β-CTX/tPINP ratio between the effective groupand the ineffective group (all P>0.05). After chemotherapy for 3 months, there were no differences in the level of tPINP compared with that before treatment in both effective group and ineffective group (all P>0.05), but the level of β-CTX decreased significantly compared with that before treatment both in effective group and ineffective group (median 0.24 vs 0.60 μg/L, 0.44 vs 0.95 μg/L, both P<0.05). The ratio of β-CTX /tPINP decreased significantly compared with that before treatment both in effective group and ineffective group (median 0.005 vs 0.012, 0.005 vs 0.011, both P<0.05). There were no differences in the level of β-CTX, tPINP and β-CTX/tPINP ratio after treatment for 6 months, 1 year and 2 years compared with that for 3 months both in effective group and ineffective group (all P>0.05)., Conclusions: Pain in bones, osteolysis and pathological fracture are the most common clinical manifestations in myeloma-related bone disease. The severity of bone disease can reflect the tumor load, but may not affect the therapeutic effect and the overall survival. The bone metabolic markers tPINP and β-CTX can be used to evaluate the severity of myeloma-related bone disease at diagnosis and to monitor the effect of treatment for bone disease.

Objective: To investigate the long-term effect of biventricular (BIV) and right ventricular apical (RVA) pacing on cardiac function in patients with high-degree atrioventricular block (AVB) and left ventricular ejection fraction(LVEF)over 35%., Methods: A total of 118 consecutive patients with high-degree AVB in six hospitals from East China between May 2009 and December 2012 were enrolled in this randomized, double-blind and parallel controlled study. Patients were randomly assigned to BIV and RVA pacing with or without LV lead on after one-week cardiac resynchronization therapy (CRT). Cardiac function including New York Heart Association(NYHA), 6 minute walking distance (6MWD), Minnesota living with heart failure (MLHF) score, LVEF, left ventricular end-diastolic volumes/diameters (LVEDV/LVEDD) and other echocardiography parameters, as well as N-terminal pro-B-type natriuretic peptide (NT-proBNP)were assessed at 6 months and 12 months., Results: A total of 114 patients were successfully implanted with CRT. Cardiac function was significantly improved after one-week BIV pacing (n=57) compared with pre-CRT: rate of patients with NYHA Ⅲ (25.44%(29/114) vs. 9.65%(11/114)), MLHF score (17.1±13.6 vs. 26.9±21.6), 6MWD ((315.4±121.8)m vs. (291.8±102.9)m) and NT-proBNP (157.0(70.0, 639.0) ng/L vs. 444.7(144.0, 1 546.0)ng/L, all P<0.05). In BIV group, 6MWD extended from (314.8±142.7)m to (332.7±117.5)m at 6 months (P<0.05), LVEF increased from (60.7±7.9)% at 1 week to (56.6±10.7)% at 6 months(P<0.05), both LVEDV and LVEDD decreased at 12 months compared with at 1 week ((116.2±39.5)ml vs. (131.4±49.6)ml and (50.2±5.6)mm vs. (52.5±6.8)mm, P<0.05). In RVA group (n=57), 6MWD increased at 6 months compared that at 1week ((342.4±109.9)m vs. (310.2±105.1)m, P<0.05), NT-proBNP was higher at 12 months than that at 1 week (349.5(191.8, 884.3)ng/L vs. 127.0(70.3, 336.7)ng/L, P<0.05). Compared with RVA group, BIV group had a bigger shrink in LVEDV decrease at 12 months was more significant in BIV group ((-16.68±24.30)ml vs. (9.09±29.30)ml, P<0.05)., Conclusions: Cardiac pacing could acutely improve the cardiac function in patients with high-degree AVB and LVEF over 35%. Improvements on cardiac function and remodeling are more significant after 12-month BIV pacing than that of RVA pacing. Clinical Trail Registry: Chinese Clinical Trial Registry, ChiCTR-TRC-10000832.

Objective: To obtain recombinant mouse collagenase ADAMTS2 (ADAM metallopeptidase with thrombospondin type 1 motif, 2) C terminal (1109-1213), and prepare the corresponding rabbit anti-ADAMTS2 polyclonal antibodies., Methods: The recombinant expression plasmid pGEX-6p-1-ADAMTS2 (1109-1213) was transformed into E.coli. The target protein was induced by IPTG and identified by mass spectrometry following affinity purification. The expressed and purified ADAMTS2 (1109-1213) protein was used to immunize New Zealand rabbits to prepare anti-ADAMTS2 polyclonal antibodies. The antibody titers were detected by ELISA and the antibody specificity by Western blotting., Results: The protein ADAMTS2 (1109-1213) was expressed in E.coli after IPTG induction, and with the purified protein, we prepared antiserum in the immunized rabbits. The titer of the antiserum reached over 1:160 000. The antiserum showed a good specificity., Conclusion: The high titer and specific rabbit anti-ADAMTS2 antibody has been prepared successfully.

To study the expression of angiotensin converting enzyme 2 (ACE2) and angiotensin (Ang) 1-7 specific receptor Mas protain in renal blood vessels of metabolic syndrome ( MS) rats and its anti-oxidative effect. A total of 80 male SD rats were divided into four groups: the normal control group (NC, the same volume of normal saline), the MS group (high fat diet), the MS + Astragali Radix group (MS + HQ, 6 g x kg(-1) x d(-1) in gavage) and the MS + Valsartan group (MS + XST, 30 mg x kg(-1) x d(-1) in gavage). After four weeks of intervention, their general indexes, biochemical indexes and blood pressure were measured; plasma and renal tissue Ang II, malondialdehyde (MDA) and superoxide demutase (SOD) levels were measured with radioimmunoassay. The protein expressions of Mas receptor, AT1R, ACE and ACE2 were detected by western blot analysis. According to the result, compared with the NC group, the MS group and the MS + HQ group showed significant increases in systolic and diastolic pressures, body weight, fasting glucose, fasting insulin, triglycerides, free fatty acid and Ang II level of MS rats (P < 0.05). The MS + XST group showed notable decreases in systolic and diastolic pressures than that of the MS group. The MS group showed significant increases in the SOD activity and NO level and decrease in the MDA level after being intervened with Astragali Radix. ACE and AT1R protein expressions in renal tissues of the MS group were higher than that in the NC group, but with lower ACE2 and -Mas receptor expressions (all P < 0.05). Compared with the MS group, the MS + HQ group showed significant increase in Mas receptor expression in renal tissues, whereas the MS + XST group showed notable decrease in AT1R (all P < 0.05). In conclusion, Astragali Radix can increase the Mas receptor expressions in renal tissues, decrease ACE expression and change local Ang II, MDA, NO and SOD in kidneys, so as to protect early damages in renal tissues.

Objective: To explore the expression of T cell immunoglobulin and mucin domain protein 3 (Tim-3) on CD56(+) NK cells in peripheral blood and its correlation with liver fibrosis indicators in patients with advanced schistosomiasis., Methods: Tim-3 expression on CD6(+) NK cells from 28 patients with advanced schistosomiasis and 30 healthy controls was determined by flow cytometry. The serum levels of IFN-y and IL-4 were detected by enzyme-linked immunosorbent assay (ELISA). Fibroscan analyzer was used for liver stiffness measurement (LSM) to determine the extent of liver fibrosis. Four serological indicators of liver fibrosis, collagen type III N-peptide (PIIP N-P), Laminin (LN), collagen IV (CIV) and hyaluronic acid (HA) were measured by the Automated Chemiluminescence Immunoassay Analyzer., Results: Flow cytometry analysis showed that Tim-3 expression on CD56(+) NK cells in advanced schistosomiasis patients was (62.3±11.4)%, significantly higher than that (52.1±6.5)% (P< 0.01). In patients with advanced schistosomiasis and the healthy controls, the levels of PIIIP N-P were (86.5±29.5) ng/mL and (22.0±7.8) ng/mL, LN (49.3±21.5) ng/mL and (20.4±6.3) ng/mL, CIV (67.5±22.3) ng/mL and (22.0±3.9) ng/mL, HA (645.9±483.1) ng/mL and (54.7±27.7) ng/mL, respectively. There were significant differences in all these indicators between the two groups (P<0.05). The levels of IFN-y were (93.9?20.1) ng/L and (107.7?24.6) ng/L, and those of IL-4 were (46.6±11.8) ng/L and (28.9±8.9) ng/L respectively in patients with advanced schistosomiasis and the healthy controls, both with significant differences (P<0.05). Spearman nonparametric correlation analysis showed that Tim-3 expression on CD56(+) NK cells in patients was positively correlated with the levels of LSM, PIIIP N-P, LN, CIV and IL-4 (r=0.528-0.834, P<0.01), but negatively with serum IFN-y (r=-0.501, P<0.01). No correlation was found with the HA level (r=0.352, P>0.05)., Conclusion: The expression of Tim-3 on peripheral CD56(+) NK cells increases in patients with advanced schistosomiasis compared with the healthy controls, and it positively correlates with the levels of LSM, PIIIP N-P, LN, CIV and IL-4, four indicators of liver fibrosis.

Tongluo Xingnao effervescent tablet (TLXNET) is a patented prescription, which comes from modified Xionggui decoction and can improve cognitive function. However, its effect on the urine metabolites and anti-dementia mechanism in the dementia model rats induced by hippocampal injection with Aβ25-35 remains unclear. The experiment focused on the changes in trajectory and inter-relationship among the urinary metabolite of rats in the blank group, Aβ25-35 hippocampal injection dementia model group and the TLXNET intervention group, in order to determine theirs characteristic metabolic markers and explain the anti-dementia effect of TLX-NET base on the change of metabolic trajectory of these bio-markers. According to the experimental results, 5, 6-indolequinone, 4-hydroxyphenyl pyruvic acid (4-HPPA), cortisol and 3-thiosulfate lactic were preliminarily identified as the characteristic metabolic markers. They mainly participate in dopamine system, glucocorticoids and energy metabolic pathways. TLXNET can apparently downregulate the disturbances of metabolic trajectory of the four bio-markers. The experiment indicates that the dementia model induced by injecting Aβ25-3 into hippocampus has its characteristic endogenous metabolic markers in urine, and ELXNET can ameliorate dementia by down-regulating the disturbances of metabolic trajectory.

Objective: To investigate the effects of angiotensin II (Ang II) antagonist telmisartan on retina vessel endothelial cell apoptosis and its impact on the ACE2-Ang-(1-7)-Mas axis in spontaneous hypertensive rats (SHR)., Methods: Thirty-six SHR 16 week-old were randomly divided into 3 groups (n = 12 each): SHR, SHRT (telmisartan 10 mg · kg-1 · d-1 by gastric gavage) and SHRTA group (telmisartan 10 mg · kg-1 · d-1 by gastric gavage plus intravenous injection of A-779 0.5 mg · kg-1 · d-1), twelve WKY rats served as normotensive control group. Systolic blood pressure was measured at pre-treatment and 8 weeks later. After 8 weeks, rats were sacrificed, the expression of ACE2 and Mas in retina were analyzed by qRT-PCR, Western blot and Immunohistochemistry, the Ang-(1-7) concentration in serum was measured by ELISA. Specimens were obtained and stained by hematoxylin and eosin, and the morphology of retina vessel was observed. Apoptosis of vessel endothelial cells were determined by using terminal deoxynucleotidyl transferase mediated dUTP nick end labeling method., Results: The systolic blood pressure of SHR, SHRT and SHRTA groups at baseline were significantly higher than age-matched WKY group (all P < 0.01). Eight weeks later, the systolic blood pressure group was significantly lower in SHRT group than in the SHR group (P < 0.01), this effect was partly reversed in SHRTA group. The retinal ACE2 mRNA and protein expression was significantly lower in SHR group than in WKY and SHRT groups (P < 0.01), which was similar between SHRT group and SHRTA group (P > 0.05). The retinal Mas mRNA and protein expression were significantly lower in SHR group compared to WKY and SHRT groups (all P < 0.01), which was significantly lower in SHRTA group than in the SHRT group (P < 0.05). ELISA results showed that serum Ang-(1-7) protein level was significantly lower in SHR group than in WKY group and SHRT group (both P < 0.05), which was lower in SHRTA group compared to SHRT group. Retinal vessel endothelial cell apoptosis was higher in SHR group than in WKY group, which could be reduced by cotreatment with telmisartan and this beneficial effect could be reversed by A-779., Conclusion: Telmisartan can reduce retinal vessel endothelial cell apoptosis via upregulating the ACE2-Ang-(1-7)-Mas axis.

Objective: To explore the therapeutic effect of grain-sized moxibustion at "Xinshu" (BL 15) and "Shenshu" (BL 23) on early-stage Alzheimer's disease (AD) in transgenosis AD mice., Methods: The genotyping of amyloid precursor protein/presenilin 1(APP/PS1I) double-transgenic AD mice were detected by PCR method. Seventeen 1.5-month female transgenic (Tg 6799) mice were randomly divided into a model group (9 cases) and a treatment group (8 cases). Nine female C57BL/6J wild-type mice with identical age and background were selected into a normal group. The treatment group was treated with grain-sized moxibustion at bilateral "Xinshu" (BI. 15) and "Shenshu" (BL 23), once a day, ten treatments were considered as one course, and total 9 courses were given. The model group and normal group were treated with stimulus such as grabbing, immobilization and non-ignited moxa cone. Morris water maze (escape latency, crossing times and dwell time in the target quadrant) was applied to evaluate the learning and memory ability. Hematoxylin-Eosin (HE) staining was used to observe morphology changes in the brains of AD mice. beta-amyloid protein 1-42 (Abeta(1-42)) in the area of prefrontal cortex and hippocampus was detected by immunohistochemical method., Results: After the treatment of grain-sized moxibustion, learning and memory ability in the treatment group was increased; compared with the model group, the escape latency was shorten, crossing times was increased, and dwell time in the target quadrant was prolonged (all P<0. 05). The crossing times and dwell time in the target quadrant in the treatment group were not significantly different from those in the normal group (both P>0.05). Compared with the normal group, the positive area and integral optical density of Abeta(1-42) in prefrontal cortex and hippocampus in the model group were increased (all P<0.01). Compared with the model group, the positive area and integral optical density of Abeta(1-42) in prefrontal cortex and hippocampus in the treatment group were reduced (P<0.01, P<0.05)., Conclusion: The grain-sized moxibustion at "Xinshu" (BL 15) and "Shenshu" (BL 23) could significantly improve the learning and memory ability in APP/PS1 double- transgenic AD mice, and inhibit the over expression and accumulation of Abeta(1-42).

Objective: To investigate the correlation between pro-B-type natriuretic peptide (NT-proBNP) and heart rate variability (HRV) in patients with diabetes., Methods: HRV and NT-proBNP levels were assessed in 58 patients with diabetes.The time domain and frequency domain indices and the level of NT-proBNP were compared between patients with normal and abnormal HRV, and the correlation of HRV index and NT-proBNP was analyzed., Results: The NT-proBNP level was significantly higher in patients with abnormal HRV than in those with normal HRV (320.2±88.3 vs 80.4±25.2 pg/ml, P<0.01).The level of NT-proBNP was negatively correlated with SDNN (r=-0.55, P<0.05), triangle index (r=-0.52, P<0.05), LF (r=-0.62, P<0.01), and VLF (r=-0.56, P<0.05)., Conclusion: NT-proBNP level is correlated with HRV, and an increased NT-proBNP indicates cardiac autonomic neuropathy in diabetic patients.

Objective: To investigate the effects of Qili Qiangxin capsules on the amino-terminal pro-brain natriuretic peptide (NT-proBNP) level and cardiac function in patients with silicosis., Methods: Hospitalized silicosis patients with heart failure were divided into treatment group (41 cases) and control group (30 cases) according to their own will. Both groups received comprehensive symptomatic treatment; in addition, the treatment group received Qili Qiangxin capsules. The treatment lasted 6 months. The observed items included NT-proBNP level, 6-minute walk test, ultrasonic cardiogram, and NYHA classification before and after treatment., Results: According to NYHA classification, the response rate was 29.27%in the treatment group and 10.00%in the control group; there was a significant difference between the two groups (P < 0.05). The average walk distance in the treatment group was increased from 150.96±73.12 m before treatment to 169.32±77.04 m after treatment, and the improvement was statistically significant (P < 0.05). The average NT-proBNP level in the treatment group was reduced from 1154.44 ± 480.79 ng/L before treatment to 494.49 ± 342.61 ng/L after treatment, and the reduction was statistically significant (P < 0.01). Left ventricular ejection fraction was significantly improved in the treatment group (P < 0.05)., Conclusion: Qili Qiangxin capsules in addition to comprehensive symptomatic treatment can significantly reduce NT-proBNP level and improve cardiac function in silicosis patients, and thereby improve patients' quality of life.

Objective: To investigate long-term efficacy and safety of ambrisentan monotherapy in patients with pulmonary arterial hypertension (PAH)., Methods: Patients with PAH who received 2.5 mg or 5 mg of ambrisentan once daily between July 10, 2011 and August 30, 2012 for at least 6 months were enrolled. The efficacy endpoints were change in exercise capacity, World Health Organization (WHO) functional class and N-terminal pro-brain natriuretic peptide (NT-proBNP) level, echocardiographic parameters. The safety endpoint was the safety of long-term ambrisentan administration, as defined by the incidence and severity of adverse events., Results: A total of 18 patients with PAH were enrolled. Mean age was (39 ± 17) years, 8 (55.6%) were female, and 11 (61.1%) patients were in WHO functional class III. The median duration of treatment was 17 months (range: 6-26 months). After treatment, the 6MWD was significantly increased[ (495 ± 97) m vs. (400 ± 91) m, P < 0.001], NT-proBNP was significantly reduced [308 (53-1 645) ng/L vs. 80(22-454) ng/L, P = 0.005], the systolic pulmonary artery was significantly decreased [(62 ± 30) mmHg vs. (82 ± 41) mmHg, P = 0.001] and left ventricular end diastolic diameter was significantly increased [(44 ± 6) mm vs. (40 ± 6) mm, P < 0.004] compared to pre-treatment. WHO functional class was improved compared with baseline in 11(61.1%) patients, stable in 7(38.9%) patients. No patient died during the treatment period. No patient was withdrawn from this study for safety reasons., Conclusions: Long-term treatment of ambrisentan can effectively improve the exercise capacity, reduce systolic pulmonary artery pressure and NT-proBNP in PAH patients. Ambrisentan is safe and well tolerated in Chinese PAH patients.

Objective: To observe the effect of Chinese herbs for stasis removing and collaterals dredging (CHSRCD) upon angiotensin-converting enzyme 2-angiotensin-(1-7)-Mas axis in the renal cortex of diabetic nephropathy rats., Methods: Totally 89 male Sprague-Dawley rats were randomly divided into the blank control group (C group, n=22), the high-glucose high-fat control group (H group, n=10), and the streptozotocin (STZ)-injecting group (n=57). The diabetes rat model (n=50) was induced by feeding high-glucose high-fat diet in combination with intraperitoneal injection of STZ, which were further divided into the model group (M group, n=24), the irbesartan group (I group, n=13), and the CHSRCD (Z group, n=13). Rats in I and Z groups were intragastrically fed with suspension of irbesartan and CHSRCD, once daily for 16 weeks. Equal volume of drinking water was administrated to rats in the rest groups. Blood glucose and 24 h urine protein quantitation were tested at four time points. And the mRNA expression of ACE2 and Mas at various time points was detected by Real-time PCR, immunohistochemical assay, and Western blot. Quantitative analyses of ACE2 and Mas protein expression were performed at the end of week 16., Results: Compared with the C group, blood glucose increased in the H and M groups (P < 0.01). It was higher in the H group (P < 0. 01). 24 h urine protein quantitation at different time points increased in the M group, and it was higher than that in the H group (P < 0.05). Compared with the M group, 24 h urine protein quantitation decreased at the end of week 8 in the I group, and at the end of week 8 and 16 in the Z group (P < 0.05). It was lower in the Z group than in the I group at the end of week 16 (P < 0.05). Compared with the C and H groups, the expression of ACE2 mRNA in the renal cortex was lower in the M group at the end of week 16 (P < 0.01). Compared with the M group, it was higher in the Z group (P < 0. 01). There was no statistical difference in the expressions of Mas mRNA at the end of week 16 between the C group and the M group (P > 0.05). It was lower in the M group than in the H group (P < 0.05). It was higher in the Z group than in the M group (P < 0.05), and higher than in the I group (P < 0.05). The expression of ACE2 and Mas protein in the M group decreased as time went by. The expression quantitation of ACE2 and Mas protein at the end of week 16 was lower in the M group than in the C group (P < 0.05). Compared with the M group, ACE2 expression of the Z group and Mas of the I and Z groups increased more significantly (P < 0. 05)., Conclusion: CHSRCD could play a role in renal protection for diabetic nephropathy rats by up-regulating the mRNA and protein expression of ACE2 and Mas, promoting the ACE2-Ang-(1-7)-Mas axis, and lowering urinary protein.

Objective: To construct a prokaryotic expression vector of the His-tagged truncated factor H-binding protein (Fhb) fragments, Fhb-N (amino acids 45-344aa) and Fhb-C (amino acids 345-644aa), of Streptococcus suis serotype 2, express it in E.coli BL21 (DE3) in order to acquire high-purity recombinant protein, and finally identify the binding activity with human serum IgG (hIgG)., Methods: Fhb-N gene and Fhb-C gene were amplified using the primers designed according to 05ZYH33 genome sequences and cloned into the expression vector pET28a⁺ to construct recombinant plasmids. The plasmids were transformed into E.coli BL21 (DE3) and induced to express by IPTG. The recombinant proteins were purified by nickel affinity chromatography and identified by Western blotting. The hIgG was purified from human serum by HiTrap protein G HP column in accordance with the manufacturer's instructions. In addition, the specific binding to hIgG was identified by Western blotting and biolayer Interferometry (BLI)., Results: The prokaryotic expression vector of His-Fhb-N and His-Fhb-C was constructed, and the target proteins were expressed, purified and identified. The specific binding activity with hIgG was identified and the binding region was found located on the Fhb-N(45-344aa)., Conclusion: His-Fhb-N can specifically bind to hIgG, which will help us to further study the role of Fhb-hIgG interaction in the pathogenesis of Streptococcus suis.

In our previous work, we found that trivalent dimethylarsinous acid (DMA(III)) have high affinity binding to cysteine residue 13 of rat hemoglobin. However, it is still unknown why arsenic intermediate metabolite DMA(III) has high binding affinity for Cysl3 but not for other cysteine residues 93, 140, 111 and 125. In order to better understand the molecular mechanism of DMA(III) with rat hemoglobin, we have done current study. So, SD rats were divided into control and arsenic-treated groups randomly. Arsenic species in lysate of red blood cells were analyzed by HPLC-ICP-MS, and then determined by a hybrid quadrupole TOF MS. In addition, trivalent DMA(III) binds to different cysteine residues in rat hemoglobin alpha and beta chains were also simulated by Molecular Docking. Only Cys13 in alpha chain is able to bind to DMA(III) from the experiment results. Cys13 of alpha chain in rat hemoglobin is a specific binding site for DMA(III), and we found that amino acids compose pockets structure and surround Cys13 (but not other cysteine residues), make DMA(III) much easy to bind cysteine 13. Taken together, the DMA(III) specific binding to Cys13 is related to spatial structure of Cys13.

Objective: To investigate the effects of exogenous hydrogen sulfide (H2S) on β-site APP cleaving enzyme 1 (BACE-1) and β-amyloid peptide (Aβ) metabolism in primary culture of neurons under high-glucose condition., Methods: The cortical neurons in primary culture under normal and high glucose (60 mmol/L) conditions for 24 h were exposed to 25, 50 and 100 µmol/L NaHS. Aβ1-42 concentration in the cell culture was measured by ELISA, and BACE-1 mRNA and protein levels were detected by fluorescent quantitative real-time PCR and Western blotting, respectively., Results: Compared with the neurons cultured in normal glucose, the neurons exposed to high glucose showed significantly increased Aβ1-42 concentration and BACE-1 mRNA and protein expressions (P<0.05). Exposure to 25, 50 and 100 µmol/L NaHS significantly decreased Aβ1-42 concentration and BACE-1 mRNA and protein expressions in the high-glucose cell culture (P<0.05)., Conclusion: Neurons exposed to high glucose exhibit increased Aβ1-42 levels and BACE-1 mRNA and protein expressions, which can be concentration-dependently decreased by NaHS.

Objective: To investigate the effect of tanshinone II(A) on the expression of different components in the renin-angiotensin system of left ventricles of renal hypertensive rats., Method: The renal hypertension model was established in rats by the two-kidney-one-clip (2K1C) method. In the experiment, all of the rats were randomly divided into four groups (n = 15 per group) before the operation: the sham-operated (Sham) group, the hypertensive model (Model) group, the low-dose tanshinone II(A) group and the high-dose tanshinone II(A) group. At 5 week after the renal artery narrowing, the third and fourth groups were administered with 35 mg kg(-1) x d(-1) and 70 mg x kg(-1) x d(-1) of tanshinone II(A), respectively. The blood pressure in rats was determined by the standard tail-cuff method in each week after the operation. After the drug treatment for 8 weeks, all the rats were put to death, and their left ventricles were separated to determine the ratio of left ventricle weight to body weight (LVW/BW), the myocardial collagen content, and the expressions of different components in myocardial RAS, including angiotensin converting enzyme (ACE), angiotensin converting enzyme 2 (ACE2), angiotensin 1-type receptor (AT1R), Mas receptor mRNA expression and angiotensin II (Ang II) and angiotensin (1-7) [Ang (1-7)] content., Result: Compared with the sham group, the hypertensive model group exhibited a markable increase in the content of Ang II and Ang (1-7) and the mRNA expressions of ACE, ACE2, AT1R and Mas (P < 0.01). However, the treatment with tanshinone II(A) showed the does dependence, inhibited left ventricle hypertrophy, decreased myocardial Ang II content and the mRNA expression of ACE and AT, R in renal hypertensive rats (P < 0. 01) , further increased the myocardial Ang (1-7) content and the mRNA expression of ACE2 and Mas (P < 0.01) , but without any change in the blood pressure of hypertensive rats., Conclusion: The treatment with tanshinone II(A) could inhibit left ventricle hypertrophy of renal hypertensive rats. Its mechanism may be partially related to the expression of different components in the renin-angiotensin system for regulating myocardial tissues.

Objective: To compare the effects of short-term and long-term thromboprophylaxis after total hip replacement on coagulation indicators in plasma sampled before and 1, 7 and 35 days post-operation., Methods: A total of 40 patients scheduled for elective total hip replacement were randomly assigned into the short-term (n = 20) or long-term (n = 20) thromboprophylaxis groups on oral rivaroxaban 10 mg once daily for 7 or 35 days. The primary efficacy hemostatic variables included thrombin-antithrombin complexes (TAT), prothrombin fragment 1+2 (F1t2), D-dimer and fibrinogen (Fib) preoperatively and at Days 1, 7 and 35 postoperatively. And ultrasonography was performed on all patients preoperatively and at days 7 and 35 postoperatively to exclude deep vein thrombosis of lower extremities., Results: None of them had deep vein thrombosis (DVT) of lower extremities. Among them, TAT, F1+2, D-dimer and Fib post-operation were higher than those preoperative baseline values. TAT and D-dimer peaked at day 1 postoperatively while the peaks of F1+2 and Fib appeared at day 7 postoperatively. At Day 35 post-operation, the levels of TAT and F1+2 in the long-term thromboprophylaxis group were significantly lower than those of the short-term thromboprophylaxis group (P < 0.05)., Conclusion: The status of hypercoagulability may sustain at least 35 days after total hip replacement. Though not completely eliminated, it can still be reduced by prolonged thromboprophylaxis. However, according to ultrasonography, the effects of short-term and long-term thromboprophylaxis on the incidence rate of DVT remain to be further explored.

Objective: To detect the expressions of neuron-specific nuclear protein (NeuN), glial fibrillary acidic protein (GFAP) and CD11b in the Aβ1-42 induced newborn rat hippocampal slices in vitro and investigate the neuroprotective effect of tanshinone IIA., Methods: Hippocampal slices were randomly divided into 7 groups: normal control group; Aβ1-42 (5.0, 0.5 μg/mL) groups; Aβ (5.0 μg/mL) + low or high (8.0 mg or 16 mg) doses of Tan IIA groups, Aβ (0.5 μg/mL) + low or high (8.0 mg or 16 mg) doses of Tan IIA groups. The expression levels of GFAP, NeuN and CD11b were detected by immunohistochemistry., Results: Compared with the control group, the expression of NeuN decreased in the two different doses of Aβ1-42 groups, especially in the high-dose group (P<0.05); while the level significantly increased after high-dose Tan IIA treatment (P<0.05). Compared with the control group, the expression levels of GFAP and CD11b increased in the two different doses of Aβ1-42 groups, and the levels in the high-dose group were the highest (P<0.05); while the levels were reduced significantly by Tan IIA treatment, the more by the high-dose Tan IIA (P<0.05)., Conclusion: Tan IIA can down-regulate the levels of GFAP and CD11b and inhibit the activity of glial cells in rat hippocampal slices induced by Aβ1-42 in vitro.

Objective: To investigate the influence of Nogo extracellular peptide residues 1-40 (NEP1-40) gene modification on the survival and differentiation of the neural stem cells (NSCs) after transplantation., Methods: NSCs were isolated from the cortex tissue of rat embryo at the age of 18 days and identified by Nestin immunofluorescence. The lentiviruses were transduced to NSCs to construct NEP1-40 gene modified NSCs. The spinal cords of 30 Sprague Dawley rats were hemisected at T9 level. The rats were randomly assigned to 3 groups: group B (spinal cord injury, SCI), group C (NSCs), and group D (NEP1-40 gene modified NSCs). Cell culture medium, NSCs, and NEP1-40 gene modified NSCs were transplanted into the lesion site in groups B, C, and D, respectively at 7 days after injury. An additional 10 rats served as sham-operation group (group A), which only received laminectomy. At 8 weeks of transplantation, the survival and differentiation of transplanted cells were detected with counting neurofilament 200 (NF-200), glial fibrillary acidic portein (GFAP), and myelin basic protein (MBP) positive cells via immunohistochemical method; the quantity of horseradish peroxidase (HRP) positive nerve fiber was detected via HRP neural tracer technology., Results: At 8 weeks after transplantation, HRP nerve trace showed the number of HRP-positive nerve fibers of group A (85.17 +/- 6.97) was significantly more than that of group D (59.25 +/- 7.75), group C (33.58 +/- 5.47), and group B (12.17 +/- 2.79) (P < 0.01); the number of groups C and D were significantly higher than that of group B, and the number of group D was significantly higher than that of group C (P < 0.01). Immunofluorescent staining for Nestin showed no obvious fluorescence signal in group A, a few scattered fluorescent signal in group B, and strong fluorescence signal in groups C and D. The number of NF-200-positive cells and MBP integral absorbance value from high to low can be arranged as an order of group A, group D, group C, and group B (P < 0.05); the order of GFAP-positive cells from high to low was group B, group D, group C, and group A (P < 0.05); no significant difference was found in the percentage of NF-200, MBP, and GFAP-positive cells between group C and group D (P > 0.05)., Conclusion: NEP1-40 gene modification can significantly improve the survival and differentiation of NSCs after transplantation, but has no induction on cell differentiation. It can provide a new idea and reliable experimental base for the study of NSCs transplantation for SCI.

Objective: To evaluate the association of major histocompatibility complex class I chain related gene A (MICA) antibodies with acute rejection (AR), chronic rejection (CR) and renal function after renal transplantation., Methods: Serum MICA antibodies were detected with ELISA before and after transplantation with also examinations of panel reactive antibodies (PRA), serum creatinine, urine, graft ultrasound, lymphocyte subsets and the pathology of graft biopsy. The study was carried out in two parts to monitor MICA antibodies in acute and chronic rejections after renal transplantation., Results: In the first part of the study 18 of the 41 recipients experienced episodes of acute rejection, and the incidence rate was markedly higher in MICA(+) group than in MICA(-) group (P<0.05). Compared with the recipients with stable renal functions, the patients with acute graft rejection showed a significantly higher positivity rate of MICA antibodies. Postoperative MICA antibody monitoring showed that MICA antibody level increased gradually 2-3 days after the occurrence of acute rejection; anti-rejection treatment lowered serum creatinine to a normal level but MICA antibodies remained positive. In the second part, 21 of 40 patients had chronic graft rejection and showed significantly higher positivity rate of MICA than the patients with stable renal functions (P<0.05). In patients with chronic rejections, the serum creatinine levels were significantly higher in MICA(+) than in MICA(-) cases (P<0.05). Graft biopsy of all MICA(+) cases showed C4d deposition., Conclusion: The status of MICA antibodies can predict the occurrence and treatment outcomes of acute rejection, and also as one of the major causes of chronic graft rejection, they affect the long-term survival of the renal grafts.

The present study investigated the effect of antiflammin-1 (AF-1) on LPS-induced IL-10 secretion from RAW264.7 cells through uteroglobin-binding protein (UGBP). Cultured RAW264.7 cells, a murine monocyte-macrophage cell line, were divided as following: control group, LPS group (1 µg/mL LPS), AF-1 group (100 μmol/L AF-1), LPS+AF-1 group (2 h of 100 μmol/L AF-1 pretreatment before LPS addition), and LPS+AF-1+anti-UGBP group (30 min of anti-UGBP antibody pretreatment before successive treatments with AF-1 and LPS). IL-10 concentration in the supernatants was detected by ELISA assay, and the level of IL-10 mRNA expression in macrophage was detected by using RT-PCR method. The results showed that AF-1 significantly increased LPS-induced IL-10 secretion in RAW264.7 cells in a dose dependent way, and up-regulated its mRNA level. Anti-UGBP antibody pretreatment attenuated the augmented effect of AF-1 on LPS-induced IL-10 secretion and gene expression. These results suggest that AF-1 promotes LPS-induced IL-10 secretion from macrophages, and this effect is mediated by UGBP.

Objective: To explore the effects of statins upon bone mineral density (BMD) and bone metabolic markers in postmenopausal women with hypercholesterolemia., Methods: A prospective study was conducted for 100 women receiving treatment from January 2011 to August 2012 and meeting the inclusion criteria of osteopenia or osteoporosis with hypercholesterolemia postmenopausal. They were randomly divided into treatment group on atorvastatin 10 mg once daily and control group. The parameters of lumbar BMD, bone resorption markers of type I collagen cross-linked C-telopeptide (CTX) , bone synthesis markers procollagen type I N-terminal peptide (PINP) were compared between two groups after half a year and one year., Results: There was an upward trend of lumbar spine BMD and PINP in the treatment group at half a year and one year compared with the control group. And two groups had significant difference (P < 0.05). Although two groups had no significant difference in all parameters at half a year, the values of lumbar spine BMD and PINP were higher in the treatment group at one year than the control group. Two groups had significant difference (P < 0.05)., Conclusions: Statins can help maintain or increase bone mass of hypercholesterolemic menopausal women through promoting bone synthesis.

Objective: To explore the expression of renin-angiotensin system component (ACE-AngII-AT1/AT2 & ACE2-Ang-(1-7)-Mas) in endometrium of polycystic ovary syndrome (PCOS) patients and normal controls., Methods: Thirty cases of normal endometrium in proliferative and secretory phases respectively were selected for the protein levels of AngII, AT1, AT2, ACE2, Ang-(1-7) and Mas through immunohistochemistry. And the specimens of proliferative endometrium from 15 PCOS patients and 15 normal controls respectively were prepared for the expressions of AT1, AT2, ACE2 and Mas mRNA through relative quantitative -polymerase chain reaction (RQ-PCR). The histological phases of endometria were confirmed by hematoxylin & eosin staining under microscope., Results: The expressions of AngII, AT1R, AT2R, ACE2, Ang-(1-7) and Mas showed periodical changes in endometrium throughout normal menstrual cycles and shared a similar trend. The expression was more pronounced in epithelial cells than that in stromal cells while it was also more dramatic in secretory phase than proliferative phase; The mRNA expressions of AT1, AT2, ACE2 and MAS were higher in PCOS endometrium than those in normal controls. Statistically significant differences existed between two groups (P < 0.01)., Conclusions: There is local existence of RAS in endometrium; Increased expressions of AT1 mRNA, AT2 mRNA, ACE2 mRNA and Mas mRNA in endometrium of PCOS may influence endometrial development and play a role in the pathological process of PCOS.

Background: Bone metastasis is common in lung cancer patients. The β isomer of the C-terminal telopeptide of type I collagen (β-CTX) and bone-specific alkaline phosphates (BAP) are regarded as important bone turnover markers in bone resorption and formation. Thus, the aims of this study are to determine the correlation of these bone turnover markers with the extent of bone metastasis of lung cancer., Methods: A total of 92 patients with bone metastasis of lung cancer from Tianjin Union Medicine Center were enrolled in this study between July 2009 and July 2012. The bone metastasis lesion was diagnosed by ECT combined with X-ray and CT imaging. The bone turnover marker levels of β-CTX and BAP were measured by ELISA., Results: Among the 92 patients, 58 cases had <3 bone-metastasis number and 34 had ≥3; 68 cases had osteolytic bone metastasis, 9 had osteoblastic bone metastasis, and 15 had mixed bone metastases. The serum levels of β-CTX and BAP were significantly higher in patients with >3 bone-metastasis number than that in patients with less than 3 (P<0.05). β-CTX was more sensitive than BAP in diagnosing lytic bone metastasis., Conclusions: β-CTX and BAP can be used as supplementary tools for diagnosing and screening bone metastasis of lung cancer.

Edible bird's nest contains lots of glycoproteins. The glycosylation inhomogeneity for glycoprotein often results in wide range of molecular weight and the difficulty for protein separation and charaterization. In this paper, proteins in the edible bird's nest were extracted using multiple extractions, and then digested by PNgase F and trypsin. The digest mixture was separated with HPLC, and peptides were identified based on MS/MS data searching. The results indicated that the extracted proteins were amount to 79.7% of total protein in the edible bird's nest. More than 20 species of peptides in the digested mixture were identified. The sequences of these peptides showed similarity with some proteins from Swiss-prot. The research indicated that deglycosylation, tryptic digestion coupled with HPLC-MS/MS is a proper strategy for characterization of proteins in the edible bird's nest.

Objective: To investigate the vasodilating effect of capsaicin (CAP) on rat mesenteric artery and its mechanism., Methods: The third branch of the superior mesenteric artery in male Sprague-Dawley rat (250-350 g) was excised, the periadventitial fat and connective tissue were removed and the mesenteric artery was dissected into 2 mm rings. Each ring was placed in a 5 ml organ bath of DMT 610M system and the tension was recorded., Results: CAP (10(-9)-10(-5) mol/L) relaxed endothelium-intact and endothelium-denuded mesenteric artery pre-constricted by phenylephrine (10(-5) mol/L), and the vasodilation in endothelium-intact mesenteric artery was stronger than that in endothelium-denuded one. Pretreatment with either L-NAME (3 X10(-4) mol/L), an inhibitor of nitric oxide synthase(NOS), or CGRP8-37 (2 X 10(-6) mol/L), an antagonist of calcitonin gene-related peptide (CGRP), for 30 min significantly attenuated the relaxation of endothelium-intact mesenteric artery induced by CAP. CGRP (10(-10)-3 X10(-8) mol/L) relaxed endothelium-intact and endothelium-denuded mesenteric artery pre-constricted by phenylephrine, and the vasodilation in endothelium-intact mesenteric artery was stronger than that in endothelium-denuded one. Substance P did not relax the mesenteric artery pre-constricted by phenylephrine., Conclusion: CAP has partial endothelium-dependent relaxation effect on rat mesenteric artery, which may be mediated by activating the endothelial NOS-NO pathway. The endothelium-independent relaxation in rat mesenteric artery induced by CAP may be mediated by CGRP.

Objective: To investigate the effects of α3 neuronal nicotinic acetylcholine receptor (nAChR) on apoptosis and p38 signal transduction pathway in SH-SY5Y cells and to assess the roles of α3 nAChR in the pathogenesis of Alzheimer's disease (AD)., Methods: The levels of α3 nAChR mRNA and protein were measured by real-time PCR and Western blot, respectively, in SH-SY5Y cells transfected with α3 nAChR siRNA. The mRNA level of bcl-2 and bax was measured by the real-time PCR. The siRNA transfected SH-SY5Y cells and control were then treated with 10 µmol/L Aβ25-35 for another 48 h, and the change in apoptotic rate and the levels of p-p38 and p38 were measured by flow cytometry and Western blot. Subsequently these SH-SY5Y cells were exposed to a blocker of p38 protein, and the apoptotic rate was measured again., Results: Compared to the controls, the expression of α3 nAChR at mRNA and protein levels in the SH-SY5Y cells transfected with α3 nAChR siRNA decreased by 95% and 86%, respectively; the mRNA levels of bax increased 2.11 times and that for bcl-2 decreased 0.53 times. The apoptotic rate was unaffected (3.40% ± 0.20%); but it increased after Aβ25-35 treatment (24.52% ± 1.59%); the level of p-p38 protein also increased by 178% in the α3 nAChR inhibited cells treated with Aβ25-35. Compared to controls, the Aβ25-35-treated SH-SY5Y cells and the Aβ25-35-treated and siRNA-transfected cells both showed a reduction in apoptosis after treatment with p38 blocker, especially in the former., Conclusion: The siRNA silencing of α3 nAChR mRNA may enhance the effect of Aβ25-35 on the cell apoptosis by increasing the levels of p38 protein and bax mRNA and decreasing the level of bcl-2 mRNA, which may play a role in the pathogenesis of AD.

Through phage display, we tried to find out whether the N-terminal fragment of glucogan-like peptide 1 receptor (nGLP-1R) still had binding activity to Exendin-4 after missing one or two gene segments. By error-prone PCR, We constructed a randomly mutated phage display peptide library with different length of the N-terminal (21-145 residues) extracellular domain of glucogan-like peptide 1 receptor (GLP-1R) from rat lung. A mutant named EP16 without binding activity was found by ELISA. Through sequence alignment we found that EP16 missed the first 20 and last 10 amino acids and the 52nd tryptophan was mutated to arginine. In order to determine why Ep16 did not show its binding ability to Exendin-4, a wild type EP16 without the first 20 and last 10 amino acids and nGLP-1R(W52R) was constructed in which the 52nd tryptophan was mutated to arginine. The contrastive analysis showed that the substitution of W52R led to a markedly reduced binding ability of EP16. The mutation of the conserved W52 could change the biologic activity of the protein. The lack of the first 20 and last 10 amino acids had no effect on its biologic activity. Therefore, the mutation of a single amino acid residue of the key sequence could change the biologic activity of the nGLP-1R.