Role and mechanism of Vγ4 T cells in impaired wound healing of rapamycin-induced full-thickness skin defects in mice / 中华烧伤杂志

Objective: To investigate the role and mechanism of Vγ4 T cells in impaired wound healing of rapamycin-induced full-thickness skin defects in mice. Methods: The experimental research methods were applied. Eighty-six C57BL/6J male mice (hereinafter briefly referred to as wild-type mice) aged 8-12 weeks were selected for the following experiments. Vγ4 T cells were isolated from axillary lymph nodes of five wild-type mice for the following experiments. Intraperitoneal injection of rapamycin for 42 mice was performed to establish rapamycin-treated mice model for the following experiments. Eighteen wild-type mice were divided into normal control group without any treatment, trauma only group, and trauma+CC chemokine ligand 20 (CCL20) inhibitor group according to the random number table (the same grouping method below), with 6 mice in each group. The full-thickness skin defect wound was made on the back of mice in the latter two groups (the same wound model below), and mice in trauma+CCL20 inhibitor group were continuously injected subcutaneously with CCL20 inhibitor at the wound edge for 3 days after injury. Another 6 rapamycin-treated mice were used to establish wound model as rapamycin+trauma group. On post injury day (PID) 3, the epidermal cells of the skin tissue around the wound of each trauma mice were extracted by enzyme digestion, and the percentage of Vγ4 T cells in the epidermal cells was detected by flow cytometry. In normal control group, the epidermal cells of the normal skin tissue in the back of mice were taken at the appropriate time point for detection as above. Five wild-type mice were used to establish wound models. On PID 3, the epidermal cells were extracted from the skin tissue around the wound. The cell populations were divided into Vγ4 T cells, Vγ3 T cells, and γδ negative cells by fluorescence-activated cell sorter, which were set as Vγ4 T cell group, Vγ3 T cell group, and γδ negative cell group (with cells in each group being mixed with B16 mouse melanoma cells), respectively. B16 mouse melanoma cells were used as melanoma cell control group. The expression of interleukin-22 (IL-22) mRNA in cells of each group was detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR), with the number of samples being 6. Thirty rapamycin-treated mice were used to establish wound models, which were divided into Vγ4 T cell only group and Vγ4 T cell+IL-22 inhibitor group performed with corresponding injections and rapamycin control group injected with phosphate buffer solution (PBS) immediately after injury, with 10 mice in each group. Another 10 wild-type mice were taken to establish wound models and injected with PBS as wild-type control group. Mice in each group were injected continuously for 6 days. The percentage of wound area of mice in the four groups was calculated on PID 1, 2, 3, 4, 5, and 6 after injection on the same day. Six wild-type mice and 6 rapamycin-treated mice were taken respectively to establish wound models as wild-type group and rapamycin group. On PID 3, the mRNA and protein expressions of IL-22 and CCL20 in the peri-wound epidermis tissue of mice in the two groups were detected by real-time fluorescence quantitative RT-PCR and Western blotting, respectively. The Vγ4 T cells were divided into normal control group without any treatment and rapamycin-treated rapamycin group. After being cultured for 24 hours, the mRNA and protein expressions of IL-22 of cells in the two groups were detected by real-time fluorescence quantitative RT-PCR and Western blotting, respectively, with the number of samples being 6. Data were statistically analyzed with independent sample t test, analysis of variance for repeated measurement, one-way analysis of variance, Bonferroni method, Kruskal-Wallis H test, and Wilcoxon rank sum test. Results: The percentage of Vγ4 T cells in the epidermal cells of the skin tissue around the wound of mice in trauma only group on PID 3 was 0.66% (0.52%, 0.81%), which was significantly higher than 0.09% (0.04%, 0.14%) in the epidermal cells of the normal skin tissue of mice in normal control group (Z=4.31, P<0.01). The percentages of Vγ4 T cells in the epidermal cells of the skin tissue around the wound of mice in rapamycin+trauma group and trauma+CCL20 inhibitor group on PID 3 were 0.25% (0.16%, 0.37%) and 0.24% (0.17%, 0.35%), respectively, which were significantly lower than that in trauma only group (with Z values of 2.27 and 2.25, respectively, P<0.05). The mRNA expression level of IL-22 of cells in Vγ4 T cell group was significantly higher than that in Vγ3 T cell group, γδ negative cell group, and melanoma cell control group (with Z values of 2.96, 2.45, and 3.41, respectively, P<0.05 or P<0.01). Compared with that in wild-type control group, the percentage of wound area of mice in rapamycin control group increased significantly on PID 1-6 (P<0.01), the percentage of wound area of mice in Vγ4 T cell+IL-22 inhibitor group increased significantly on PID 1 and PID 3-6 (P<0.05 or P<0.01). Compared with that in rapamycin control group, the percentage of wound area of mice in Vγ4 T cell only group decreased significantly on PID 1-6 (P<0.05 or P<0.01). Compared with that in Vγ4 T cell only group, the percentage of wound area of mice in Vγ4 T cell+IL-22 inhibitor group increased significantly on PID 3-6 (P<0.05 or P<0.01). On PID 3, compared with those in wild-type group, the expression levels of IL-22 protein and mRNA (with t values of -7.82 and -5.04, respectively, P<0.01) and CCL20 protein and mRNA (with t values of -7.12 and -5.73, respectively, P<0.01) were decreased significantly in the peri-wound epidermis tissue of mice in rapamycin group. After being cultured for 24 hours, the expression levels of IL-22 protein and mRNA in Vγ4 T cells in rapamycin group were significantly lower than those in normal control group (with t values of -7.75 and -6.04, respectively, P<0.01). Conclusions: In mice with full-thickness skin defects, rapamycin may impair the CCL20 chemotactic system by inhibiting the expression of CCL20, leading to a decrease in the recruitment of Vγ4 T cells to the epidermis, and at the same time inhibit the secretion of IL-22 by Vγ4 T cells, thereby slowing the wound healing rate.

Clinical value of a differentiation prediction model for invasive lung adenocarcinoma / 中华肿瘤杂志

Objective: To investigate the value of predicting the degree of differentiation of pulmonary invasive adenocarcinoma (IAC) based on CT image radiomics model and the expression difference of immunohistochemical factors between different degrees of differentiation of lesions. Methods: The clinicopathological data of patients with pulmonary IAC confirmed by surgical pathology in the Affiliated Huai'an First People's Hospital to Nanjing Medical University from December 2017 to September 2018 were collected. High-throughput feature acquisition was performed for all outlined regions of interest, and prediction models were constructed after dimensionality reduction by the minimum absolute shrinkage operator. Receiver operating characteristic curve was used to assess the predictive efficacy of clinical characteristic model, radiomics model and individualized prediction model combined with both to identify the degree of pulmonary IAC differentiation, and immunohistochemical expressions of Ki-67, NapsinA and TTF-1 were compared between groups with different degrees of IAC differentiation using rank sum test. Results: A total of 396 high-throughput features were extracted from all IAC lesions, and 10 features with high generalization ability and correlation with the degree of IAC differentiation were screened. The mean radiomics score of poorly differentiated IAC in the training group (1.206) was higher than that of patients with high and medium differentiation (0.969, P=0.001), and the mean radiomics score of poorly differentiated IAC in the test group (1.545) was higher than that of patients with high and medium differentiation (-0.815, P<0.001). The differences in gender (P<0.001), pleural stretch sign (P=0.005), and burr sign (P=0.033) were statistically significant between patients in the well and poorly differentiated IAC groups. Multifactorial logistic regression analysis showed that gender and pleural stretch sign were related to the degree of IAC differentiation (P<0.05). The clinical feature model consisted of age, gender, pleural stretch sign, burr sign, tumor vessel sign, and vacuolar sign, and the individualized prediction model consisted of gender, pleural stretch sign, and radiomic score, and was represented by a nomogram. The Akaike information standard values of the radiomics model, clinical feature model and individualized prediction model were 54.756, 82.214 and 53.282, respectively. The individualized prediction model was most effective in identifying the degree of differentiation of pulmonary IAC, and the area under the curves (AUC) of the individualized prediction model in the training group and the test group were 0.92 (95% CI: 0.86-0.99) and 0.88 (95% CI: 0.74-1.00, respectively). The AUCs of the radiomics group model for predicting the degree of differentiation of pulmonary IAC in the training group and the test group were 0.91 (95% CI: 0.83-0.98) and 0.87 (95% CI: 0.72-1.00), respectively. The AUCs of the clinical characteristics model for predicting the degree of differentiation of pulmonary IACs in the training and test groups were 0.75 (95% CI: 0.63-0.86) and 0.76 (95% CI: 0.59-0.94), respectively. The expression level of Ki-67 in poorly differentiated IAC was higher than that in well-differentiated IAC (P<0.001). The expression levels of NapsinA, TTF-1 in poorly differentiated IAC were higher than those in well-differentiated IAC (P<0.05). Conclusions: Individualized prediction model consisted of gender, pleural stretch sign and radiomics score can discriminate the differentiation degree of IAC with the best performance in comparison with clinical feature model and radiomics model. Ki-67, NapsinA and TTF-1 express differently in different degrees of differentiation of IAC.

Correlation of the single nucleotide polymorphisms rs34349826 and rs6521 of the LHB gene with male infertility in Chinese men / 中华男科学杂志

<p><b>Objective</b>To study the relationship of the single nucleotide polymorphisms (SNP) rs34349826 (c.104 A>G) and rs6521 (c.114 C>G) of the luteinizing hormone beta-subunit (LHB) gene with male infertility in Chinese men.</p><p><b>METHODS</b>This case-control study included 405 males with primary infertility (the infertility group) and 424 normal fertile men (the control group), the former again divided into subgroups of oligospermia, severe oligozoospermia and azoospermia according to the sperm concentration. Clinical data were collected from all the subjects and genomic DNA obtained from their peripheral blood for genotyping rs34349826 and rs6521 of the LHB gene by Sequence MassArray. We analyzed the correlation of male infertility with the SNPs of the two loci using the logistic regression model as well as its association with their haplotype combination with the SHEsis online software.</p><p><b>RESULTS</b>There were statistically significant differences between the control and infertility groups in the semen volume (［3.51 ± 1.36］ vs ［3.74 ± 1.71］ ml, P <0.05), sperm concentration (［79.21 ± 61.60］ vs ［27.37 ± 30.80］ ×10⁶/ml, P <0.01), percentage of progressively motile sperm (［39.40 ± 9.64］ % vs ［11.90 ± 14.72］ %, P <0.01), and levels of serum luteinizing hormone (LH) (［3.29 ± 1.39］ vs ［6.25 ± 4.83］ IU/L, P <0.01) and follicle-stimulating hormone (FSH) (［4.56 ± 2.31］ vs ［15.64 ± 17.03］ IU/L, P <0.01). Logistic regression analysis revealed no correlation between male infertility and the genotypes of the rs34349826 and rs6521 loci of the LHB gene, and similar results were found in the subgroups of the infertile males. SHEsis analysis on the haplotypes of the rs34349826 and rs6521 loci showed the GG genotype combination to be a protective factor against male infertility.</p><p><b>CONCLUSIONS</b>The rs34349826 and rs6521 loci of the LHB gene were not related to male infertility, which can be further confirmed by larger-sample studies. The GG genotype combination is a protective factor against male infertility.</p>

Key techniques for precision cultivation of nitrogenous fertilizer of pollution-free ginseng / 中国中药杂志

Planting pollution-free farmland is the main mode of industrialization of ginseng cultivation, fine management of nitrogen fertilizer ginseng pollution-free farmland cultivation technology system is one of the key factors. In order to investigate the effect of nitrogenous fertilizer on the accumulation of ginseng biomass and saponins synthesis in vegetative growth stage, two-years-old ginsengs were used as test materials in this study. The test materials were cultivated by Hoagland medium with different nitrogen concentration (0，10，20，40 mg·L⁻¹) for 40 days. During the cultivation, photosynthetic rate was measured four times. After 40 days cultivation, chlorophyll content, stem diameter and the spatiotemporal expression of saponin synthesis related genes PgHMGR and PgSQE were tested. The results showed that there were significant differences in the photosynthetic rate and chlorophyll content among different nitrogen concentrations. The relative expression level of PgHMGR gene and PgSQE gene in root, stem and leaves of ginseng were different. Ginseng seedlings cultivated by 20 mg·L⁻¹ nitrogen possess the highest photosynthetic rate and chlorophyll content, while PgHMGR and PgSE showed the highest gene expression level. The optimal nitrogen concentration for the growth of 2-years-old ginseng might be 20 mg·L⁻¹ with 57.14 g ammonium nitrate each plant or pure 20.00 mg nitrogen each plant. It is concluded that this concentration is the most suitable concentration for the ginsenoside synthesis. Pollution-free ginseng with fine nitrogen fertilizer cultivation is conducive to the production of high quality and efficient ginseng medicinal materials. It lays a theoretical foundation for the rational fertilization and environment-friendly sustainable ecological ginseng planting industry.

The prevalence of post traumatic stress disorder in an army unit and its relationship with functional gastrointestinal disorders / 解放军医学杂志

Objective To determine the prevalence of post traumatic stress disorder (PTSD) in the army officers and soldiers (AOSs),and identify its relationship with functional gastrointestinal disorders (FGIDs).Methods PTSD and FGIDs were diagnosed based on the PTSD checklist--civilian (PCL-C) and Rome Ⅲ Modular Questionnaire respectively,the overlaps of PTSD and FGIDs and their correlation were diagnosed.The correlation of PTSD with traumatic and stressful events was investigated using Trauma History and Stressful Event Screening Questionnaire.The coexistence and relationship of PTSD and FGIDs were analyzed.Results Of 927 AOSs,33 were diagnosed with PTSD.The prevalence of PTSD was 3.56％.FGIDs were identified in 435 subjects and the incidence of FGIDs was 46.93％.Among 33 AOSs with PTSD,28 were diagnosed as having FGIDs and the prevalence of FGIDs was 84.85％,which was significantly higher than that of non-PTSD group (45.53％,P＜0.05).Moreover,the FGIDs group had a higher prevalence of PTSD,compared with the non-FGID group (6.43％ vs.1.02％,P＜0.05).Cyclic vomiting syndrome (CVS,33.33％),unspecified functional bowel disorder (24.24％),functional bloating (18.18％) and functional anorectal pain (18.18％) were the four most frequent FGIDs in PTSD AOSs.Multiple regression analysis showed PTSD was the risk factor for CVS (OR=9.118),functional anorectal pain (OR=3.373),functional bloating (OR=4.772),irritable bowel syndrome (OR=3.438),rumination syndrome (OR=16.033),functional vomiting (OR=10.329),functional dysphagia (OR=9.891)(P＜0.05).CVS (OR=4.063),the number of traumatic (OR=1.159) and stress events (OR=1.401) were the risk factors for PTSD in AOSs (P＜0.05).Conclusions PTSD and FGID interact as risk factor each other.The prevalence of PTSD differs significantly in different FGIDs.CVS is the most frequent FGID in PTSD AOSs and risk factor for PTSD,which deserves more concerns.

Effect of Modified Sini San on Cell Morphology and Expression of 5-Hydroxytryptamine Transporter in Hippocampal CA1 Region of Depression Rats Induced by Adolescent and Post-adult Stress / 广州中医药大学学报

Objective To investigate the cell morphology and expression of 5-hydroxytryptamine transporter(5-HTT) in hippocampal CA1 region of depression rats induced by adolescent and post-adult stress,and to observe the inter-vention effect of modified Sini San (MSS). Methods One hundred and thirty-two Wistar rats were randomly divided into blank group,model group,JSS group,and fluoxetine group,33 rats in each group. And then the rats in each group were randomly subdivided into adolescent group, adult group, post-adult group acaccording to the age day, 11 rats in each subgroup. Age day 44,56 and 78 were used as the sampling time points for adolescent group,adult group,post-adult group respectively. Chronic unpredictable mild stress(CUMS)rat model was used. From day 21 to 44 and from day 57 to 78, the rats were modeled and given medication, but from day 44 to 55, the rats were fed normally. The rat general condition and body mass of various groups were observed,the cell morphology of hippocampal CA1 region was observed by hematoxylin-eosin (HE) staining , and the distribution of 5-hydroxytryptamine transporter (5-HTT)positive cells in CA1 region of hippocampus was observed by immunohistochemical staining. Results The general condition of the rats at different age stages in the model group was poor,while that in MSS group and fluoxetine group was improved obviously. The body mass of rats at different age stages in the model group was obviously decreased (P<0.01 compared with the blank group). After adulthood stage,the body mass of rats in model group, MSS group, and fluoxetine group was lower than that of the blank group(P < 0.01), but there was no difference between the 3 groups (P > 0.05). In aspect of cell morphological manifestation in hippocampal CA1 region, rats in the adolescent model group had more deeply-staining atrophy neurons, with unclear hyperchromatic nucleus and cytoplasm. The morphological manifestations in modeled rats at adult stage and post-adulthood stage showed progressive aggravation,manifested as a large amount of neurons stained deeply with unclear nucleus and cytoplasm, and a small amount of glial cells proliferated. Compared with the model group at the same stage,the neuronal atrophy and deeply staining decreased in fluoxetine group and MSS group. The average optical density value of 5-HTT expression in the model group was decreased significantly at the adult stage and after adulthood stage(P<0.05 or P<0.01 compared with the blank group). Compared with the model group, the average optical density value of 5-HTT expression in MSS group after adulthood stage, and in the fluoxetine group at the adult stage and after adulthood stage were increased (P<0.05 or P<0.01). Conclusion Rats suffering CUMS in adolescence presents depressive behavior, and post-adult stimulation can aggravates depression. 5-HTT expression in hippocampus may be an important pathway for MSS to achieve the therapeutic efficacy.

Effect of β-asarone on differential protein expression in brain tissue of APPswe/PSldE9 double transgenic mice / 第二军医大学学报

Objective To investigate the effect of β-asarone on differential protein expression in brain tissue of APPswe/PS1dE9 double transgenic mice, and to explore its mechanism in treatment of Alzheimer disease (AD). Methods The animals were divided into normal control group (C57BL/6J mice), model group (APPswe/PS1dE9 mice) and β-asarone treatment group (APPswe/PS1dE9 mice), with ten mice in each group. In a period of 90 days, the mice in β-asarone treatment group were administered with β-asarone by intragastric gavage (15 mg/[kg·d]), and the mice in normal control and model groups were administered with equal doses of normal saline. The learning and memory abilities of mice were detected by Morris water maze test. The expression of β-amyloid precursor protein (APP) in brain tissues was detected by immunohistochemistry. Proteomics analysis of brain tissues was performed by isobaric tags for relative and absolute quantification (iTRAQ). The expression of differential protein H2A and H2B was identified by Western blotting. Results Compared with the model group, the escape latency and the first latency time required to find the escaped platform of mice in the β-asarone treatment group were significantly shortened (P<0.05), the across-platform times were significantly increased (P<0.05), the expression of APP was significantly decreased (P<0.05), and the expressions of H2A 1-H, H2B 2-E and H2B 1-F/J/L were significantly decreased (P<0.05). Conclusion β-Asarone plays a therapeutic role by intervening the modification of histone, which might be one of the mechanisms to improve learning and memory abilities injured by the toxicity of β-amyloid peptide.

Huoxue Anxin Recipe () promotes myocardium angiogenesis of acute myocardial infarction rats by up-regulating miR-210 and vascular endothelial growth factor / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the microRNAs (miRNAs) expression profile of acute myocardial infarction (AMI) rats and the regulating effects of Huoxue Anxin Recipe (, HAR) on angiogenesis-related miRNAs and genes.</p><p><b>METHODS</b>Forty-five Wistar rats were randomly assigned to 3 groups according to a random number table: sham, AMI, and AMI+HAR groups (15 in each group). AMI rats were established by ligation of the left descending coronary artery. HAR was intragastrically administered to rats of the AMI+HAR group for successive 21 days since modeling, meanwhile the same volume of 0.9% normal saline was administered to rats of the sham and AMI groups. Doppler echocardiography was used for noninvasive cardiac function test. Hematoxylin and eosin staining was used to observe the histopathological change. miRNAs expression profile was detected by quantitative realtime polymerase chain reaction (qRT-PCR). The mRNA and protein expressions of vascular endothelial growth factor (VEGF), and a target gene of miR-210 was further detected by qRT-PCR and Western blot, respectively. The microvessels density of myocardium was evaluated by CD31 immunostaining.</p><p><b>RESULTS</b>Compared with the sham group, ejection fraction (EF) and fractional shortening (FS) values were decreased significantly in the AMI group (P<0.01), while the infarction area and the interstitial collagen deposition were increased obviously. As for the AMI+HAR group, EF and FS values were increased significantly (P<0.05 vs. AMI group), and the infarction area was reduced and the interstitial collagen deposition were alleviated significantly. Total of 23 miRNAs in the AMI group expressed differently by at least 1.5 folds compared with those in the sham group; 5 miRNAs in the AMI+HAR group expressed differently by at least 1.5 folds compared with those in the AMI group. Among them, miR-210 was low in the AMI group and high in the AMI+HAR group. The relative mRNA and protein expressions of VEGF were decreased significantly in the AMI group (P<0.05 vs. sham group), and increased significantly in the AMI+HAR group (P<0.01 vs. AMI group). CD31 expression area and optical intensity were decreased significantly in the AMI group (P<0.05 vs. sham group), and increased significantly in the AMI+HAR group (P<0.01 vs. AMI group).</p><p><b>CONCLUSIONS</b>HAR could reduce the infarction area, alleviate the interstitial fibrosis and improve the cardiac function of AMI rats. Those effects could be related to promoting myocardium angiogenesis of HAR by up-regulating miR-210 and VEGF.</p>

Effect of traditional Chinese medicine on coronary heart disease with phlegm and blood stasis syndrome / 中国中药杂志

Coronary heart disease (CHD) with pathological characteristics of atherosclerotic coronary artery and myocardial ischemia is more likely to cause phlegm and blood stasis. The current study showed high morbidity and severity of phlegm and blood stasis syndrome in CHD, which has attracted increasing attention. This paper introduced the knowledge of traditional Chinese medicine (TCM) about the CHD with phlegm and blood stasis syndrome. On the basis of modern medical studies, we found a close relationship between phlegm and blood stasis syndrome and blood fat, hemorheology, oxygen free radical, blood coagulation function, inflammation, sugar metabolism and other related gene expression changes. We reviewed TCM therapies and prescriptions and found that TCM therapies achieved a good clinical effect through the understanding of etiology and pathogenesis and differentiation in organs. Besides, phlegm and blood stasis removing prescription relieved the objective indicators, which would have a good development prospects.

Relationship between calcium overload and myocardial ischemia reperfusion injury and intervention strategy of Chinese herbal medicine / 中国中药杂志

The recent report from National Cardiovascular Center shows that cardiovascular diseases account for more than 40% of disease deaths among residents, so it has become the first cause of death among the residents in our country, and the mortality of coronary heart disease is increasing year by year. Revascularization can quickly open the clogged blood vessels and restore coronary blood supply, so it is an important approach for the treatment of coronary heart disease. However, the revascularization can not terminate the pathological development of coronary heart disease because it is just a local treatment method. In addition, a series of reperfusion injuries after revascularization would seriously restrict its treatment effect for coronary heart disease. Myocardial ischemia reperfusion injury is a complex pathological process, which is closely related to oxygen free radicals, calcium overload and energy metabolism disorder. The calcium overload can be seen during reperfusion in the myocardial cells, and it can cause further damages to the myocardial cells through various mechanisms. Calcium overload is a common pathway of myocardial necrosis and apoptosis, so prevention and treatment of calcium overload is an important method to prevent ischemia reperfusion. The commonly used calcium channel blockers for preventing calcium overload has made great progress, all of which can act on L-type calcium channel of vascular smooth muscle to inhibit calcium overload. However, their clinical application was restrained to a certain extent due to the single target and great side effects. Traditional Chinese medicine (TCM) is a great treasure, and many drugs in TCM have similar effects with calcium antagonists, so the development and application of such drugs would be an important task for contemporary TCM doctors to make up for the deficiency of Western medicine.

Impact of sleep deprivation on coronary heart disease and progress in prevention and treatment with traditional Chinese medicines / 中国中药杂志

Sleep deprivation (SD) has been taken as an independent predictor for cardiovascular risks, which was closely related to the increased morbidity and mortality in coronary heart disease (CHD). In this article, after reviewing the impact of modern medical method sleep deprivation on CHD and studies on principle method recipe medicines for preventing and treating CHD, the authors observed the autonomic nerve dysfunction, hormonal metabolism dysfunction, endothelial dysfunction and inflammatory responses after sleep deprivation, which can cause or aggravate CHD. On the basis of the traditional Chinese medicine theories of "heart dominating the blood and vessels and the mind", the authors considered that traditional Chinese medicines can tonify heart and soothe the nerves, reducing all of the risk factors through multi-target and multi-pathway, and improve sleep and decrease the risk factors caused by sleep deprivation, which provides a new idea for the prevention and treatment of CHD.

Cordblood-Based High-Throughput Screening for Deafness Gene of 646 Newborns in Jinan Area of China

OBJECTIVES: Infants with slight/mild or late-onset hearing impairment might be missed in universal newborn hearing screening (UNHS). We identified the mutation hot spot of common deaf gene in the newborns in Jinan area population by screening the mutation spot with neonate cord blood, in order to make clear whether the neonate cord blood for screening is feasible. METHODS: Six hundred and forty-six newborns were subjected to both UNHS and genetic screening for deafness by using neonate cord blood. The newborn genetic screening targeted four deafness-associated genes, which were commonly found in the Chinese population including gap junction beta-2 protein (GJB2), gap junction beta-3 protein (GJB3), solute carrier family 26 member 4 (SLC26A4), and mtDNA 12S rRNA. The most common 20 spot mutations in 4 deaf genes were detected by MassARRAY iPLEX platform and mitochondrial 12S rRNA A1555G and C1494T mutations were sequenced using Sanger sequencing. RESULTS: Among the 646 newborns, 635 cases passed the UNHS and the other 11 cases (1.7%) did not. Of the 11 failures, two cases were found to carry homozygous GJB2 p.R143W pathogenic mutation, one case was found to have heterozygous GJB2 235delC mutation, and another one case carried heterozygous GJB3 p.R180X pathogenic mutation. Six hundred and thirty-five babies passed the newborn hearing screening, in which 25 babies were identified to carry pathogenic mutations, including 12 heterozygotes (1.9%) for GJB2 235delC, eight heterozygotes (1.3%) for SLC26A4 IVS7-2A>G, one heterozygote (0.2%) for p.R409H, two homozygotes (0.3%) for m.1494C>T, and two homozygotes (0.3%) for m.1555A>G. CONCLUSION: Newborn genetic screening through the umbilical cord blood for common deafness-associated mutations may identify carriers sensitive to aminoglycoside antibiotic, and can effectively prevent or delay hearing loss occurs.

Promoting development of new traditional Chinese medicine by combination disease-syndrome and multi-objective optimization research in prevention and treatment of cardiovascular disease / 中国中药杂志

Differences in theories, application forms and evaluation standards about curative effect between traditional Chinese medicine and modern medicine lead to not only question safty and effectiveness but also hinder development and internationalization of traditional Chinese medicine. Combination of common problems in traditional Chinese new drug registration with experiences in research on traditional Chinese new drugs of prevention and treatment of coronary heart disease elucidate application value about theory of disease-syndrome combination and multi-objective optimization in several ways such as the indications positioning, preparation process optimization, preclinical efficacy evaluating and clinical assessmenting of efficacy and analysis development prospect.

Expounding the functions of qi in TCM based on the effect mitochondria / 中国中西医结合杂志

Qi in TCM is the most essential substance that makes up the body and maintains life activities. All vital substances in the body are transformed by constant motion and changes of qi. Qi in TCM mainly means full of functions. What is the basic material attribute of qi? We don't have a systematic study on it. Therefore, we combined the achievement of modern medicine, and explored further from the origin, functions, pathogeneses, and therapies of mitochondria and qi. Surprisingly, we found out that the origin of mitochondria was similar to that of qi. They are tiny substance constituting the human body. Secondly, the function of mitochondria is similar to that of qi. When the disorder of qi and mitochondria occurs, similar vital signs occur or the same reactions occur. These results suggested that the basic material attribute of qi might be mitochondria.

Effect of danlou tablet on arrhythmia model rats induced by transient myocardial ischemia/ reperfusion / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the effect of Danlou Tablet (DT) on arrhythmia model rats induced by transient myocardial ischemia/reperfusion (I/R).</p><p><b>METHODS</b>Totally 45 healthy Wistar rats were randomly divided into 3 groups, the sham-operation group, the model group, and the DT group, 15 in each group. Rats in the sham-operation group and the model group were administered with distilled water by gastrogavage at the daily dose of 0.1 mL/kg. Rats in the DT group was administered with 0.53 g/mL DT suspension by gastrogavage at the daily dose of 0.1 mL/kg. All medication was lasted for 10 successive days. The myocardial I/R experiment was performed at 1 h after the last gastrogavage. ECG was performed before ligation and at I/R. The jugular arterial blood pressure of all rats was measured during the whole course. ST segment changes were observed at each time point of I/R. The ventricular fibrillation, the premature ventricular, the number and the duration of ventricular tachycardia within 30 min reperfusion were also observed. Activities of Na(+)-K+ ATPase and Ca2+ ATPase in the myocardium homogenate were detected as well.</p><p><b>RESULTS</b>The jugular arterial blood pressure and the heart rate were slightly lower in the DT group than in the model group, but with no statistical difference (P > 0.05). Compared with the sham-operation group, the degree of ST segment was obviously elevated in the model group at 0, 5, and 7 min (P < 0.05). It was significantly lower in the DT group than in the model group (P < 0.01). ST seg ment was more elevated at 5 min than at 0 min in the model group, but the degree of ST segment elevation was still obviously lower in the DT group than in the model group (P < 0.05). There was no statistical difference in the degree of ST segment elevation at 7 min between the two groups (P > 0.05). At 0 min when the decrement of ST segment exceeded one half the ischemia, there was no statistical difference in the degree of myocardial ischemia between the model group and the DT group (P > 0.05). Compared with the model group, the incidence of fatal and nonfatal ventricular fibrillation, the frequency and duration of ventricular tachycardia and premature ventricular beats were obviously lessened, and activities of Na(+)-K+ ATPase and Ca(2+)-ATPase increased (all P < 0.05).</p><p><b>CONCLUSIONS</b>DT could significantly protect arrhythmias induced by transient I/R. Its effect might be related to lowering the degree of myocardial ischemia, and increasing ion transport channel related enzyme activities.</p>

Measurement of penile length in children and its significance / 中华男科学杂志

Nowadays, more and more parents are paying increasing attention to the penile length of their children. At present, the methods of measuring penile length mainly include manual measurement and ultrasonography. The former can be used to measure the flaccid, stretched and erected penile lengths, and its use for measuring the stretched penile length has been internationally accepted for its precise definition, unified description, and high repeatability. The latter, as a new method, is being gradually accepted for its imaging visualization and measurement accuracy. This article reviews different measurements of penile length in the mainstream literature of recent years, with an analysis of their advantages and disadvantages.

Huoxue anxin recipe alleviated peroxidation damage of acute myocardial infarction rats by regulating iNOS/eNOS imbalance: an experimental research / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the protective mechanism of Huoxue Anxin Recipe (HAR) on peroxidation damage of acute myocardial infarction (AMI) rats.</p><p><b>METHODS</b>The AMI rat model was established by occluding the left anterior descending coronary artery. Compound Danshen Dripping Pill (CDDP) was used as the positive control. CDDP and HAR were administered to rats for 7 successive days since modeling. The heart function was detected using color Doppler echocardiography. Activities of induced nitric oxide synthase (iNOS), endothelial nitric oxide synthase (eNOS), total superoxide dismutase (tSOD) activity, and contents of malondialdehyde (MDA) were detected by ultraviolet spectrophotometer method.</p><p><b>RESULTS</b>Compared with the sham-operation group, ejection fraction (EF) and fraction shortening (FS) rate significantly decreased in the model group (P < 0.01). Compared with the model group, EF and FS rate significantly increased in the HAR group, showing statistical difference (P < 0.05). There was no statistical difference in activities of serum iNOS, eNOS, or tSOD among all groups (P > 0.05). Compared with the sham-operation group, iNOS activities and MDA contents significantly increased in the myocardial tissue of the model group (P < 0.01), activities of eNOS and tSOD significant decreased (P < 0.01). Compared with the model group, iNOS activities in the myocardial tissue, MDA contents both in serum and the myocardial tissue significantly decreased (P < 0.05), activities of eNOS and tSOD significantly increased in the HAR group (P < 0.05). There was no statistical difference in each index between the CDDP group and the HAR group (P > 0.05).</p><p><b>CONCLUSIONS</b>HAR could significantly improve cardiac functions of AMI rats. Its roles might be associated with regulating imbalanced iNOS/eNOS expressions and alleviating peroxidation damage of the myocardial tissue.</p>

Experimental study on attenuating ischemic injury of acute myocardial infarction rats by Huoxue Anxin Recipe / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the protection effects and the mechanisms of Huoxue Anxin Recipe (HAR) on acute myocardial infarction (AMI) rats.</p><p><b>METHODS</b>The AMI Wistar rat model was prepared by ligating the left anterior descending coronary artery. By taking elantan long as the positive control drug, HAR was extracted from Chinese herbs by modern pharmacological methods and composed according to theories of Chinese medicine (CM). The medication time started from the day of modeling to the 21st day of the modeling. The heart function, the morphological changes of the heart, changes in the mRNA and protein levels of toll like receptor 4 nuclear factor kappa B tumor necrosis factor alpha (TLR4-NFkappaB-TNFalpha) pathway were observed.</p><p><b>RESULTS</b>Compared with the sham-operation group, the ejection fraction (EF) and left ventricular fractional shortening (FS) significantly decreased (P < 0.01), the left ventricular intemal dimension end-diastolic (LVIDd) and left ventricular internal dimension end-systolic (LVIDs) significantly increased (P < 0.01), the mRNA and protein levels of TLR4-NFkappaB-TNFalpha channel significantly increased in the model group (P < 0.01). The infarction in the front wall of the left ventricle was obviously seen, accompanied with severe inflammatory cell infiltration and collagen deposition in model group. Compared with the model group, the EF and FS significantly increased, LVIDd and LVIDs significantly decreased in the positive control group, the high and low dose HAR groups (P < 0.05, P < 0.01). The infracted area of the front wall of the left ventricle was obviously contracted. The inflammatory cell infiltration and collagen deposition were obviously alleviated. In the high and low dose HAR groups, the mRNA and protein expression levels of TLR4-NFkappaB-TNFalpha significantly decreased (P < 0.05, P < 0.01), but no inhibition was found in the positive control group.</p><p><b>CONCLUSIONS</b>HAR could significantly improve the morphological structures and functional abnormality induced by myocardial ischemia in AMI rats. Its effects was correlated with inhibiting TLR4-NFkappaB-TNFalpha pathway.</p>

Salvianolic acid B inhibits the TLR4-NFκB-TNFα pathway and attenuates neonatal rat cardiomyocyte injury induced by lipopolysaccharide / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the role of the TLR4-NFκB-TNFα inflammation pathway on: lipopolysaccharide (LPS)-induced neonatal rat cardiomyocyte injury and the possible protective effects of salvianolic acid B (Sal B).</p><p><b>METHODS</b>Wistar rat (1-2 days old) cardiomyocytes were isolated and cultured. Sal B 10(-5)mol/L, 10(-6)mol/L and 10(-7)mol/L were pre-treated for 6 h in the culture medium. LPS (1 μg/mL) was added to mol/the culture medium and kept for 6 h to induce inflammation injury. The concentration of lactate dehydrogenase (LDH) in the supernatant was detected by spectrophotometry. The concentrations of tumor necrosis factor α (TNFα) and heat shock protein 70 (HSP70) in the supernatant were detected by enzyme linked immunosorbent assay. The protein expressions of toll, such as receptor 4 (TLR4) and nuclear factor kappa B (NFκB) were detected by immunohistochemistry. The mRNA expressions of TLR4 and NFκB were detected by real-realtime reverse transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>(1) The concentrations of LDH and: TNFα in the LPS control group were significantly higher than those in the control group (561.41±67.39 U/L and 77.94±15.08 pg/mL, versus 292.13±26.02 U/L and 25.39±16.53 pg/mL, respectively, P<0.01, P<0.05). Compared with the LPS control group, the concentrations of LDH and TNFα were significantly decreased in the Sal B 10(-5)mol/L pre-treated group (451.76±83.96 U/L and 34.00±10.38 pg/mL, respectively, P<0.05). (2) The TLR4 and NFκB protein expression area in the LPS control group were significantly higher than those in the control group (1712.41±410.12 μm(2) and 2378.15±175.29 μm(2), versus 418.62±24.42 μm(2) and 1721.74±202.87 μm(2), respectively, P<0.01). The TLR4 and NFκB protein expression internal optical density (IOD) values in the LPS control group were also significantly higher than those in the control group (3.06±0.33 and 7.20±1.04, versus 0.91±0.21 and 4.24±0.48, respectively, P<0.05 and P<0.01). Compared with the LPS control group, the TLR4 and NFκB protein expression areas were significantly decreased in the Sal B 10(-5)mol/L pre-treated group (1251.54±133.82 μm(2) and 1996.37±256.67 μm(2), respectively, P<0.05), the TLR4 and NFκB protein expression IOD values were also significantly decreased in the Sal B 10(-5)mol/L pre- mol/pretreated group (1.92±0.28 and 5.17±0.77, respectively, treated P<0.05). (3) The TLR4 and NFκB mRNA expressions (2(-ΔΔ)CT value) in the LPS control group were significantly higher than those in the control group (3.16±0.38 and 5.03±0.43 versus 1.04±0.19 and 1.08±0.21, respectively, P<0.01). Compared with the LPS control group, the TLR4 and NFκB mRNA expressions (2(-ΔΔ) -CT value) were significantly decreased in the Sal B 10(-5)mol/L pre- mol/pretreated group (1.34±0.22 and 1.74±0.26, respectively, treated P<0.05). The concentration of HSP70 did not show any <statistical differences in all groups (P>0.05).</p><p><b>CONCLUSIONS</b>The TLR4-NFκB-TNFα pathway was quickly activated: and was independent of HSP70 in the early phase of neonatal cardiomyocyte injury induced by LPS. The protective effects of Sal B may be through inhibiting the TLR4-NFκB-TNFα pathway and are dose-dependent.</p>

Experimental study on the induced differentiation of human amnion mesenchymal cells into osteoblasts / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the feasibility of inducing differentiation of the human amniotic mesenchymal cells (hAMCs) into osteoblasts in vitro, so as to provide the seed cells for bone tissue engineering.</p><p><b>METHODS</b>The hAMCs were isolated from abandoned human amnion and cultured in osteogenic media to induce the osteogenic differentiation in vitro. After hAMCs were induced by osteogenic media for 15 days, morphological observation, immunocytochemistry and western blot were used to study the cellular morphology and expression of alkaline phosphatase (ALP), type I collagen, osteopontin and osteocalcin.</p><p><b>RESULTS</b>The primary cultured hAMCs had long spindle shape or irregular shape, which were distributed evenly. The cells were usually suheultured in 5 or 7 days. After subculture, the cells became larger. After cultured by osteogenic media for 15 days, the hAMCs were detected to express ALP, osteocalcin and osteopontin, and secrete type I collagen.</p><p><b>CONCLUSIONS</b>The hAMCs are isolated, cultured and amplified easily in vitro. The induced differentiated cells by osteogenic media have typical osteoblast morphological and functional characteristics, which can be used as seed cells for bone tissue engineering.</p>