Tongxie Yaofang Regulates Expression of NKG2DL to Enhance Anti-tumor Effect of NK Cells in Colon Cancer under Chronic Stress / 中国实验方剂学杂志

ObjectiveTo observe the effect of Tongxie Yaofang on the function of tumor-related natural killer （NK） cells under chronic stress and explore the possible molecular mechanism. MethodFifty SPF-grade BABL/C male mice were randomized into normal， model， and low-， medium-， and high-dose （6.825， 13.65， and 27.3 g·kg-1， respectively） Tongxie Yaofang groups， with 10 mice in each group. Other groups except the blank group were subjected to 7 days of chronic restraint stress， and then forced swimming and tail suspension tests were carried out to evaluate the modeling performance. After the successful modeling， rats in Tongxie Yaofang groups were administrated with low-， medium-， and high-doses of Tongxie Yaofang by gavage， while those in the other groups were administrated with normal saline by gavage. After 14 days， each group of mice was inoculated with subcutaneous colon cancer to establish the model of colon cancer under chronic stress. The pathological changes of the tumor tissue in each group of mice were observed using hematoxylin-eosin （HE） staining. The content of CD49b-positive cells in the peripheral blood and tumor tissue of mice was measured by flow cytometry. Enzyme-linked immunosorbent assay （ELISA） was employed to measure the content of molecules associated with NK cell activation in the peripheral blood. Western blot was employed to determine the protein levels of major histocompatibility complex class Ⅰ polypeptide-related sequences A and B （MICA+MICB） and UL-16-binding protein 1 （ULBP1） in the tumor tissue. ResultCompared with the normal group， the model group showed a decrease in 5-hydroxytryptamine （5-HT） content and an increase in corticosterone （CORT） content in the serum （P<0.05）. Compared with the model group， Tongxie Yaofang increased the 5-HT content and decreased the CORT content （P<0.05， P<0.01）. Compared with the normal group， the modeling increased the tumor volume and weight （P<0.05）， while Tongxie Yaofang inhibited such increases with no statistical significance. The tumor cells in the model group presented neat arrangement， irregular shape， uneven size， obvious atypia， common nuclear division， and small necrotic area， and blood vessels were abundant surrounding the tumor cells. Compared with the model group， Tongxie Yaofang groups showed sparse arrangement of tumor cells， different degrees of patchy necrosis areas in the tumor， and karyorrhexis， dissolution， and nuclear debris in the necrotic part. Compared with the normal group， the model group showed reduced CD49b-positive cells in the peripheral blood and tumor tissue （P<0.01）. Compared with the model group， Tongxie Yaofang increased CD49b-positive cells （medium dose P<0.01， high dose P<0.05， P<0.01）. Compared with the normal group， the modeling lowered the serum levels of granzymes-B （Gzms-B）， perforin （PF）， interferon （IFN）-γ， and tumor necrosis factor （TNF）-α （P<0.05， P<0.01）. Compared with the model group， low-dose Tongxie Yaofang elevated the serum levels of PF， Gzms-B， and TNF-α （P<0.05， P<0.01）， and medium-dose Tongxie Yaofang elevated the serum levels of Gzms-B， PF， IFN-γ， and TNF-α （P<0.05， P<0.01）. In addition， high-dose Tongxie Yaofang elevated the serum levels of PF， IFN-γ， and TNF-α （P<0.01）. Compared with the normal group， the model group presented down-regulated protein level of ULBP1 （P<0.05）. Compared with the model group， low-， medium-， and high-dose Tongxie Yaofang up-regulated the protein level of ULBP1 （P<0.05， P<0.01）， and medium- and high-dose Tongxie Yaofang up-regulated the protein level of MICA+MICB （P<0.05， P<0.01）. ConclusionTongxie Yaofang may promote NK cell activation by up-regulating the expression of MICA+MICB and ULBP1， thereby delaying the progression of colon cancer under chronic stress.

Tongxie Yaofang Regulates T Lymphocyte Subsets to Improve Immune Microenvironment of Colorectal Cancer Under Chronic Stress / 中国实验方剂学杂志

ObjectiveTo explore the effect of Tongxie Yaofang on the immune microenvironment of colorectal cancer in mice under chronic stress and the underlying mechanism. MethodA total of 40 male SPF BABL/C mice were randomized into normal group， stress group， Tongxie Yaofang group （13.65 g·kg-1）， and Tongxie Yaofang-stress group （13.65 g·kg-1）， with 10 in each group. Chronic restraint stress was induced in mice and administration （ig） of Tongxie Yaofang began after 7 days of stress. On the 14th day， forced swim and tail suspension tests were used to examine the behavioral changes of mice after stress and the subcutaneous colorectal tumor was implanted in each group of mice. The effect of this prescription on the body mass and tumor volume of mice was observed. After the last administration， mouse serum and tumor samples were collected. The content of T lymphocytes （CD3+， CD4+， CD8+， and CD4+/CD8+） in tumor was detected by immunohistochemistry and flow cytometry and levels of corticosterone （CORT） in peripheral blood， and interleukin （IL）-2， interferon-γ （IFN-γ）， IL-6， and IL-10 in the serum were determined by enzyme-linked immunosorbent assay （ELISA）. The protein expression of inhibitor of nuclear factor-κB（IκB） kinase α/β （IKKα/β）， nuclear factor-κB （NF-κB）α （IκBα）， NF-κB p65， and phosphorylated （p）-NF-κB p65 was measured by Western blot. ResultCompared with the normal group， the stress group had large tumor volume （P<0.05）， low content of CD3+， CD4+， and CD4+/CD8+ （P<0.05， P<0.01）， high content of CD8+， low content of T helper 1 （Th1）-secreted IFN-γ （P<0.05）， high content of T helper 2 （Th2）-secreted IL-10 （P<0.05） and CORT （P<0.05）， high protein expression of p-NF-κB p65， NF-κB p65， and IKKα/β （P<0.05）， and low protein expression of IκBα （P<0.05）. Compared with the normal group， the Tongxie Yaofang group showed slow tumor growth， high content of CD3+， CD4+， and CD4+/CD8+ （P<0.01）， low content of CD8+ （P<0.05）， high content of Th1-secreted IL-2 and IFN-γ （P<0.05）， low content of Th2-secreted IL-6 and IL-10 （P<0.05）， low content of CORT， low protein expression of p-NF-κB p65， NF-κB p65， and IKKα/β （P<0.05）， and high protein expression of IκBα （P<0.01）. Tongxie Yaofang-stress group demonstrated slower tumor growth， higher content of CD3+， CD4+， and CD4+/CD8+ （P<0.01）， smaller content of CD8+ （P<0.05）， higher content of IL-2 and IFN-γ （P<0.05）， lower content of IL-6， IL-10 （P<0.05）， and CORT （P<0.05）， lower protein expression of p-NF-κB p65， NF-κB p65， and IKKα/β （P<0.05，P<0.01）， and higher protein expression of IκBα （P<0.01） than the stress group. ConclusionTongxie Yaofang can delay the growth of colorectal cancer under chronic stress and alleviate the deterioration of the immune microenvironment， possibly by inhibiting NF-κB signaling pathway， regulating the function of T lymphocyte subsets， and thus suppressing the secretion of pro-inflammatory factors.

Inhibitory Effect and Mechanism of Sishenwan-containing Serum on Aerobic Glycolysis in Human Colon Cancer Cells / 中国实验方剂学杂志

ObjectiveTo explore the effect and mechanism of Sishenwan-containing serum on aerobic glycolysis in human colon cancer HCT116 cells. MethodCell counting kit-8 （CCK-8） was used to detect the cell viability of colon cancer HCT116 cells after treatment with Sishenwan-containing serum （2.5%， 5%， and 10%） for 24， 48， 72 h. The concentration of lactic acid， the content of intracellular glucose， and the activity of hexokinase （HK） and fructose-6-phosphate kinase （PFK） in the cell culture medium were detected by the micro-method. The content of glucose transporter 1 （GluT1） mRNA was detected by Real-time quantitative polymerase chain reaction （Real-time PCR）. The protein expression of GluT1 and methyltransferase-like 3 （MettL3） was detected by Western blot. The expression of GluT1 in cells was detected by immunofluorescence and the level of N6-methyladenosine （m6A） RNA methylation was detected by colorimetry. ResultCompared with the normal serum， 2.5%， 5%， and 10% Sishenwan-containing serum had no significant effect on the viability of HCT116 cells at 24 h， while 10% Sishenwan-containing serum showed a significant inhibitory effect on the viability of HCT116 cells at 48 h （P<0.05）. Hence， 10% Sishenwan-containing serum was used in subsequent experiments， and the intervention time was 48 h. Compared with the normal serum， 10% Sishenwan-containing serum could reduce lactate production （P<0.05）， down-regulate glucose uptake （P<0.05）， and blunt the activities of HK and PFK， the key rate-limiting enzymes of glycolysis （P<0.05）. Meanwhile， 10% Sishenwan-containing serum could decrease the expression of GluT1 protein （P<0.01） and mRNA （P<0.05） and reduce the proportion of cells expressing GluT1 （P<0.01）. Compared with the normal serum， Sishenwan-containing serum also decreased the protein content of MettL3 （P<0.05） and the methylation level of m6A RNA （P<0.01）. ConclusionSishenwan can inhibit glycolysis in colon cancer cells， and its inhibitory mechanism may be related to reducing MettL3 overexpression， inhibiting m6A RNA methylation， and down-regulating GluT1 and the activities of intracellular aerobic glycolysis-related enzymes such as HK and PFK.

Clinical characteristics and outcomes of severe community- and hospitalacquired pneumonia / 中国感染与化疗杂志

Objective To explore the clinical characteristics and prognostic factors of severe community-acquired pneumonia (SCAP) and severe hospital-acquired pneumonia (SHAP). Methods Clinical data of patients with severe pneumonia were reviewed and analyzed retrospectively. Multivariate logistic regression analysis was conducted to investigate the potential prognostic factors. Results A total of 70 SCAP cases and 110 SHAP cases were identified. SHAP patients showed higher proportion of surgical operation (P＜0.001), radiotherapy and/or chemotherapy (P=0.006) within 1 month, higher Charlson comorbidity index (P=0.001), and more malignancies (P＜0.001) than SCAP patients, but fewer failed organs (P=0.048), lower proportion of heart failure (P=0.003), and lower level of BNPmax (P=0.037) than SCAP. SHAP patients showed more total days of hospital stay than SCAP patients [33.0 (25.0, 43.3) days versus 14.0 (5.8, 28.3) days, P＜0.001 ], and higher 180-day mortality than SCAP patients (44.6％ versus 27.5％, P=0.047). The 180-day survival rate was significantly different between SCAP and SHAP patients (χ2=4.009, P=0.045). Multivariate analysis indicated that APACHE-II score (OR=20.449, P=0.002), cancer (OR=6.183, P=0.039), maximum D-dimer level (OR=13.841, P=0.008), and renal replacement therapy (RRT) (OR=19.456, P=0.003) were associated with the outcomes of SCAP patients. Two or more organ failure (OR=6.308, P=0.028), ventilator-associated pneumonia (OR=4.491, P=0.005), and RRT (OR=11.456, P=0.002) were associated with the outcomes of SHAP patients. Conclusions SCAP doesn't show significant difference from SHAP in in-hospital mortality or 30-day mortality. SHAP is associated with higher 180-day mortality than SCAP.

Analysis and countermeasures of suspected medical devices adverse events of hospital / 中国医学装备

Objective:To sort out cases of suspected medical device adverse events in Subei hospital, analysis for the reasons of adverse events, and propose measures to ensure the safe clinical use of medical devices.Methods: From the National Adverse Drug Reaction Monitoring Centre database, 300 cases of suspicious medical device adverse events were selected from cities and provinces. The number of categories of adverse events involving medical devices and the main reasons resulting adverse events were analyzed by SPSS.Results: The main reasons resulting the medical device adverse events were quality-related issues of the product, biological incompatibility existing between the patient and the physical device, and improper operation or use.Conclusion: The enterprises of medical devices should product on the basis of GMP strictly to ensure the quality of product. The medical institutions should ask hypersensitivity of patients actively and operate on the basis of SOP strictly to improve the work of monitoring and reporting.

MUSCULAR METABOLISM IN LOW SE CHILDREN / 西安交通大学学报(医学版)

A series of etiological investigations were made on muscular metabolism in low Se children and rats, including (1) low Se environment and low Se state in children, (2) moasurement of some biochemical indexes concerning muscular metabolism (selenium, creatine, ereatinine, phosphate, CPK, etc.), (3) Se supplemented experiment on children in low Se state, (4) experimental study on Se deficiency and Se supplemented dietary in rats, (5) patho-histological findings on the muscular tissues of children with KBD, and (6) effect of Se deficiency on muscular metabolism in children. The results show that there may be a causal relationship between Se deficiency in environment and muscular metabolism di- sturbance in low Se children. A hyphothesis on the disturbance of energy metabolism in mitochondria caused by Se deficiency is put forward that Se deficiency may blunt creatinephosphate synthesis in mitochondria and cause lesions in the mitochondrial structure, which was different from the antioxidant fiction. There is a marginal deficiency in Se for children in this paper, i.e. to take more than 20 ?tg Se/day is necessary. We suggest that muscular atrophy in low Se children be precisely designated by the name of "endemic Seresponsive muscular dystrophy".