Lipophosphatidic acid(LTA)was used to stimulate Mac-T cells,and the expression lev-els and phosphorylation levels of key proteins of nuclear factor-κB(NF-κB)and mitogen-activated protein kinase(MAPK)signaling pathway and the expression levels of upstream key action factors TLR4 and MyD88 proteins were detected by Western blot,and EDU assay was used to detect cell proliferation levels and flow cytometry was used to detect apoptosis.The results showed that acti-vation of GPR120 significantly decreased the phosphorylation levels of LTA-induced NF-κB(P65 and IκBα)(P＜0.01)and MAPK(JNK,ERK,p38)(P＜0.01)in Mac-T cells;inhibition of GPR120 was able to upregulate LTA-induced NF-κB(p65 and IκBα)in Mac-T cells(P＜0.01)and MAPK(JNK,ERK,p38)phosphorylation levels(P＜0.01);and activation of GPR120 significantly allevia-ted LTA-induced upregulation of TLR4 and MyD88(P＜0.01);inhibition of GPR120 significantly exacerbated LTA-induced upregulation of TLR4 and MyD88(P＜0.05);LTA stimulation led to a trend of diminished Mac-T cell proliferation and significantly increased apoptosis,whereas activa-tion of the GPR120 gene significantly increased cell activity(P＜0.01),promoted cell proliferation and significantly reduced apoptosis(P＜0.05)thereby alleviating the damage to Mac-T cells by LTA;LTA stimulation led to a highly significant increase in apoptosis(P＜0.01).In contrast,acti-vation of the GPR120 gene significantly reversed the increase in the apoptosis rate of Mac-T cells induced by LTA(P＜0.01),while inhibition of the GPR120 gene enhanced the apoptosis-promo-ting effect of LTA(P＜0.05),indicating that activation of the GPR120 gene attenuated the in-crease of apoptosis rate caused by LTA-induced inflammatory Mac-T cells.The results suggest that GPR120 can regulate inflammation by mediating TLR4 and MyD88 expression to inhibit NF-κB/MAPK inflammatory pathway activation and can promote cell proliferation.

Objective To investigate the status quo of pain catastrophizing(PC)in the patients with di-abetic peripheral neuropathic pain(DPNP),and to analyze the influencing factors to provide reference for for-mulating clinical preventive intervention strategies.Methods A total of 206 patients with DPNP admitted and treated in the People's Hospital of Guangxi Zhuang Autonomous Region were selected as the research sub-jects by convenience sampling method.The general data questionnaire,Numerical Rating Scale(NRS),Pain Catastrophizing scale(PCS),Perceived Social Support Scale(PSSS)and diabetes distress scale(DDS)were used to conduct the investigation.Results The incidence rate of PC in 206 cases of DPNP patients was 44.66％(92/206),and the total score of PCS was(30.10±5.16)points.The results of multiple linear regres-sion analysis showed that the gender,duration of diabetes(≥10 years),multiple drug use,number of compli-cations(＞5),NRS score,PSSS score and scores of DDS dimensions were the main influencing factors of PC(all P＜0.05),which could explain 92.3％of the total variation of PC.Conclusion The PC incidence rate in the patients with DPNP is high.Clinical healthcare workers should pay attention to the evaluation of PC in these patients,and formulate the scientific and effective targeted intervention measures according to the main influen-cing factors to help the patients to reduce the pain burden in order to reduce the level of PC.

Objective:To investigate the interactive effect of mismatch repair (MMR) protein status and adverse clinicopathological features on prognosis of stage Ⅰ-Ⅲ colon cancer.Methods:The retrospective cohort study was conducted. The clinicopathological data of 1 650 patients with colon cancer of stage Ⅰ-Ⅲ who were admitted to 7 hospitals in China from January 2016 to December 2017 were collected. There were 963 males and 687 females, aged 62(53,71)years. Patients were classified as 230 cases of MMR deficiency (dMMR) and 1 420 cases of MMR proficiency (pMMR) based on their MMR protein status. Observation indicators: (1) comparison of clinicopathological characteristics between patients of different MMR protein status; (2) analysis of factors affecting the survival outcomes of patients of dMMR; (3) analysis of factors affecting the survival outcomes of patients of pMMR; (4) interaction analysis of MMR and adverse clinicopathological features on survival outcomes. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the independent t test. Measurement data with skewed distribution were represented as M( Q1, Q3), and comparison between groups was conducted using the Mann-Whitney U test. Count data were described as absolute numbers, and comparison between groups was conducted using the chi-square test or Fisher exact probability. Comparison of ordinal data was conducted using the Mann-Whitney U test. The random forest interpolation method was used for missing values in data interpolation. Univariate analysis was conducted using the COX proportional risk regression model, and multivariate analysis was conducted using the COX stepwise regression with forward method. The coefficient of multiplication interaction effect was obtained using the interaction term coefficient of COX proportional risk regression model. Evaluation of additive interaction effects was conducted using the relative excess risk due to interaction ( RERI). Results:(1) Comparison of clinicopathological characteristics between patients of different MMR protein status. There were significant differences in age, T staging, the number of lymph node harvest, the number of lymph node harvest <12, high grade tumor between patients of dMMR and pMMR ( P<0.05). (2) Analysis of factors affecting the survival outcomes of patients of dMMR. Results of multivariate analysis showed that T staging, N staging, the number of lymph node harvest <12 were independent factors affecting the disease-free survival (DFS) of colon cancer patients of dMMR ( hazard ratio=3.548, 2.589, 6.702, 95% confidence interval as 1.460-8.620, 1.064-6.301, 1.886-23.813, P<0.05). Age and N staging were independent factors affecting the overall survival (OS) of colon cancer patients of dMMR ( hazard ratio=1.073, 10.684, 95% confidence interval as 1.021-1.126, 2.311-49.404, P<0.05). (3) Analysis of factors affecting the survival outcomes of patients of pMMR. Results of multivariate analysis showed that age, T staging, N staging, vascular tumor thrombus were independent factors affecting the DFS of colon cancer patients of pMMR ( hazard ratio=1.018, 2.214, 2.598, 1.549, 95% confidence interval as 1.006-1.030, 1.618-3.030, 1.921-3.513, 1.118-2.147, P<0.05). Age, T staging, N staging, high grade tumor were independent factors affecting the OS of colon cancer patients of pMMR ( hazard ratio=1.036, 2.080, 2.591, 1.615, 95% confidence interval as 1.020-1.052, 1.407-3.075, 1.791-3.748, 1.114-2.341, P<0.05). (4) Interaction analysis of MMR and adverse clinicopathological features on survival outcomes. Results of interaction analysis showed that the multiplication interaction effect between the number of lymph node harvest <12 and MMR protein status was significant on DFS of colon cancer patients ( hazard ratio=3.923, 95% confidence interval as 1.057-14.555, P<0.05). The additive interaction effects between age and MMR protein status, between high grade tumor and MMR protein status were significant on OS of colon cancer patients ( RERI=-0.033, -1.304, 95% confidence interval as -0.049 to -0.018, -2.462 to -0.146). Conclusions:There is an interaction between the MMR protein status and the adverse clinicopathological features (the number of lymph node harvest <12, high grade tumor) on prognosis of colon cancer patients of stage Ⅰ-Ⅲ. In patients of dMMR, the number of lymph node harvest <12 has a stronger predictive effect on poor prognosis. In patients of pMMR, the high grade tumor has a stronger predictive effect on poor prognosis.

Protein aggregation is a critical issue in the production of biopharmaceuticals. During protein production, transport and storage, various factors can lead to protein aggregation. With the in-depth study, different ways of protein aggregation and various influencing factors were identified. This includes physical and chemical factors, translation modifications and protein structure. Since protein aggregation exerts major impact on the activity and homogeneity of proteins, it is of great importance to study the ways of protein aggregation and how to control it to obtain high-quality proteins. The review focuses on three ways of protein aggregation, namely 3D domain swapping, salt bridge formation, and oxidative stress, as well as methods to control protein aggregation during protein production, transport and storage. This may facilitate reducing the loss caused by the formation of protein aggregation and improving the purity and homogeneity of protein in research and commercial production.
Protein Aggregates ; Proteins/chemistry* ; Oxidative Stress

Objective:To investigate the role of cholinergic anti-inflammatory pathway in the regulation of peptide transporter 1 (PepT1) expression in small intestinal epithelium of septic rats by Ghrelin.Methods:One hundred adult male Sprague-Dawley (SD) rats were randomly divided into sham operation group, sepsis group, sepsis+vagotomy group, sepsis+Ghrelin group, and sepsis+vagotomy+Ghrelin group, with 20 rats in each group. In the sham operation group, the cecum was separated after laparotomy, without ligation and perforation. In the sepsis group, the rats received cecal ligation puncture (CLP). In the sepsis+vagotomy group, the rats received CLP and vagotomy after laparotomy. In the sepsis+Ghrelin group, 100 μmol/L Ghrelin was intravenously injected after CLP immediately. The rats in the sepsis+vagotomy+Ghrelin group received CLP and vagotomy at the same time, then the Ghrelin was intravenously injected immediately with the same dose as the sepsis+Ghrelin group. Ten rats in each group were taken to observe their survival within 7 days. The remaining 10 rats were sacrificed 20 hours after the operation to obtain venous blood and small intestinal tissue. The condition of the abdominal intestine was observed. The injury of intestinal epithelial cells was observed with transmission electron microscopy. The contents of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in serum and small intestinal tissue were detected by enzyme-linked immunosorbent assay (ELISA). The brush border membrane vesicle (BBMV) was prepared, the levels of mRNA and protein expression of PepT1 in the small intestinal epithelium were detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and Western blotting.Results:All rats in the sham operation group survived at 7 days after operation. The 7-day cumulative survival rate of rats in the sepsis group was significantly lower than that in the sham operation group (20% vs. 100%, P < 0.05). The cumulative survival rate of rats after Ghrelin intervention was improved (compared with sepsis group: 40% vs. 20%, P < 0.05), but the protective effect of Ghrelin was weakened after vagotomy (compared with sepsis+Ghrelin group: 10% vs. 40%, P < 0.05). Compared with the sham operation group, in the sepsis group, the small intestine and cecum were dull red, the intestinal tubules were swollen and filled with gas, the intestinal epithelial cells were seriously injured under transmission electron microscopy, the levels of TNF-α and IL-1β in serum and small intestinal were significantly increased, and the expression levels of PepT1 mRNA and protein in the small intestinal epithelium were significantly decreased. It indicated that the sepsis rat model was successfully prepared. After vagotomy, the intestinal swelling and gas accumulation became worse in septic rats, leading to the death of all rats. Compared with the sepsis group, the abdominal situation in the sepsis+Ghrelin group was improved, the injury of intestinal epithelial cells was alleviated, the serum and small intestinal TNF-α and IL-1β were significantly decreased [serum TNF-α (ng/L): 253.27±23.32 vs. 287.90±19.48, small intestinal TNF-α (ng/L): 95.27±11.47 vs. 153.89±18.15, serum IL-1β (ng/L): 39.16±4.47 vs. 54.26±7.27, small intestinal IL-1β (ng/L): 28.47±4.13 vs. 42.26±2.59, all P < 0.05], and the expressions of PepT1 mRNA and protein in the small intestinal epithelium were significantly increased [PepT1 mRNA (2 -ΔΔCt): 0.66±0.05 vs. 0.53±0.06, PepT1 protein (PepT1/GAPDH): 0.80±0.04 vs. 0.60±0.05, both P < 0.05]. Compared with the sepsis+Ghrelin group, after vagotomy in the sepsis+vagotomy+Ghrelin group, the effect of Ghrelin on reducing the release of inflammatory factors in sepsis rats was significantly reduced [serum TNF-α (ng/L): 276.58±19.88 vs. 253.27±23.32, small intestinal TNF-α (ng/L): 144.28±12.99 vs. 95.27±11.47, serum IL-1β (ng/L): 48.15±3.21 vs. 39.16±4.47, small intestinal IL-1β (ng/L): 38.75±4.49 vs. 28.47±4.13, all P < 0.05], the up-regulated effect on the expression of PepT1 in small intestinal epithelium was lost [PepT1 mRNA (2 -ΔΔCt): 0.58±0.03 vs. 0.66±0.05, PepT1 protein (PepT1/GAPDH): 0.70±0.02 vs. 0.80±0.04, both P < 0.05], and the injury of small intestinal epithelial cells was worse. Conclusion:Ghrelin plays a protective role in sepsis by promoting cholinergic neurons to inhibit the release of inflammatory factors, thereby promoting the transcription and translation of PepT1.

Objective:To investigate the changes in the expression of toll like receptor 4 (TLR4), pentamerin-3 (PTX3) and Apelin in the blood of patients with sepsis and their relationship with the condition and prognosis.Methods:From March 2015 to March 2020, 82 patients with sepsis in the First People′s Hospital of Chenzou were retrospectively selected, including 22 patients in septic shock group, 34 patients in severe sepsis group and 26 patients in general sepsis group. 82 healthy people in the same period were selected as the control group. The expression of serum TLR4, PTX3, Apelin, Sequential Organ Dysfunction Score (SOFA), Acute Physiology and Chronic Health Score System Ⅱ (APACHE Ⅱ) were measured and compared. The relationship between the expression of serum TLR4, PTX3 and Apelin and the scores of SOFA and APACHE Ⅱ in patients with sepsis were analyzed. The general data and the expression of serum TLR4, PTX3 and Apelin in patients with sepsis with different prognosis (28 day survival and death) were counted. The influencing factors of prognosis in patients with sepsis were observed, and the predictive significance of serum TLR4, PTX3 and Apelin on the prognosis of patients with sepsis was analyzed.Results:Comparison of serum TLR4, PTX3 and Apelin level in the groups: septic shock group>severe sepsis group>general sepsis group>control group ( P<0.05); The serum level of TLR4, PTX3 and Apelin in patients with high SOFA and APACHE Ⅱ scores were higher than those in patients with low SOFA and APACHE Ⅱ scores ( P<0.05); Pearson correlation showed that serum level of TLR4, PTX3 and Apelin was positively correlated with the SOFA and APACHE Ⅱ scores of sepsis patients ( P<0.05); The levels of serum TLR4, PTX3, Apelin and the scores of SOFA and APACHE Ⅱ in the dead patients with sepsis were higher than those in survival patients ( P<0.05); Logistic regression analysis found that serum level of TLR4, PTX3, Apelin, SOFA score, and APACHE Ⅱ score were all important risk factors for 28 day death of sepsis patients ( P<0.05); The receiver operating characteristic (ROC) curve showed that the sensitivity and specificity of combined blood indexes (TLR4, PTX3 and Apelin) in predicting the prognosis of patients with sepsis were 85.71% and 85.25%. Conclusions:The expression of serum TLR4, PTX3 and Apelin in patients with sepsis can be significantly increased, which is related to the patient′s condition and prognosis and can provide a certain basis for clinical diagnosis and treatment.

Objective:To investigate the role of LncRNA-TUG1 in the injury of intestinal epithelial cells induced by lipopolysaccharide (LPS).Methods:LPS was used to treat HIEC-6 human intestinal epithelial cells for 24 h to construct a sepsis injury model. Whole transcriptome RNA sequencing was used to analyze the expression changes of mRNA, microRNA and lncRNA in HIEC-6 cells after LPS treatment. Real-time fluorescence quantitative (qRT-PCR) and Western blot was performed to detect the expression changes of lncRNA-TUG1, microRNA-132-3p (miR-132-3p), SIRT1 mRNA and SIRT1 protein in HIEC-6 cells after LPS treatment. The expression levels of LncRNA-TUG1, miR-132-3p and SIRT1 were artificially changed by in vitro transfection. qRT-PCR and Western blot were used to confirm the regulatory effect of lncRNA-TUG1 on microRNA-132-3p and SIRT1. CCK-8 and flow cytometry were used to analyze the effects of LncRNA-TUG1, miR-132-3p and SIRT1 on the proliferation and apoptosis of HIEC-6 cells. The dual luciferase report analysis was used to verify the targeting relationship between LncRNA-TUG1, miR-132-3p and SIRT1. Statistical analysis was performed using SPSS 17.0, and differences between the two groups were compared using independent sample t test. Results:RNA sequencing results showed that the expressions of lncRNA-TUG1 and SIRT1 were decreased in HIEC-6 cells after LPS treatment ( t=3.26, P<0.05 and t=2.55, P<0.05), but the expression of miR-132-3p was increased ( t=4.12, P<0.05). In vitro cell experiments, the expression of lncRNA-TUG1 and SIRT1 were decreased in HIEC-6 cells treated with LPS ( t=5.69, P<0.05 and t=5.712, P<0.05), while the expression of miR-132-3p was increased ( t=3.88, P<0.05). Overexpression of lncRNA-TUG1 increased the proliferation rate ( t=6.55, P<0.05) and decreased the apoptosis rate ( t=3.94, P<0.05) of LPS-treated cells. Upregulation of lncRNA-TUG1 decreased the expression of miR-132-3p ( t=4.66, P<0.05), and increased the mRNA and protein levels of SIRT1 ( t=3.91, P<0.05). Transfection of miR-132-3P mimic could inhibit the mRNA ( t=4.08, P<0.05) and protein levels of SIRT1. In LPS-treated cells, the cells co-transfected with miR-132-3pmimic and siRNA-SIRT1 had a lower proliferation rate ( t=4.55, P<0.05 and t=5.67, P<0.05) and a higher apoptosis rate ( t=3.90, P<0.05 and t=4.22, P<0.05) than those transfected with only pcDNA3.1-lncRNA-TUG. Conclusions:lncRNA-TUG1 may act as a ceRNA to regulate miR-132-3p/SIRT1, therefore alleviating HIEC-6 cell injury caused by LPS. Intervention of lncRNA-TUG1/miR-132-3p/SIRT1 regulatory pathway may become a potential strategy to prevent sepsis-induced intestinal mucosal damage.

Objective: To identify the definition of heat wave based on mortality risk assessment in different regions of China. Methods: Daily mortality (from China Information System for Disease Control and Prevention) and meteorological data (from National Meteorological Information Center in China) from 66 counties with a population of over 200 000 were collected from 2006-2011. With the consideration of climate type and administrative division, China was classified as seven regions. Firstly, distributed lag non-linear model (DLNM) was used to estimate community-specific effects of temperature on non-accidental mortality. Secondly, a multivariate meta-analysis was applied to pool the estimates of community-specific effects to explore the region-specific temperature threshold and the duration for definition of heat wave. Results: We defined regional heat wave of Northeast, North, Northwest, East, Central and Southwest China as being two or more consecutive days with daily mean temperature higher than or equal to the P₆₄, P₇₁, P₈₅, P₆₇, P₇₅ and P₇₇ of warm season (May to October) temperature, respectively, while the thresholds of temperature were 21.6, 23.7, 24.3, 25.7, 28.0 and 25.3 ℃. The heat wave in South China was defined as five or more consecutive days with daily mean temperature higher than or equal to the P₉₃ (30.4 ℃) of warm season (May to October) temperature. Conclusion The region-specific definition of heat wave developed in our study may provide local government with the guidance of establishment and implementation of early heat-health response systems to address the negative health outcomes due to heat wave.

Objective To finding out the characters of vascular remolding after the establishment of native arteriovenous fistula on the wrist,and exploring the influential factors.Methods Doppler ultrasound was used to monitor the diameter of cephalic vein,brachial artery,radial artery and ulnar artery at the time before the surgery and one day,one week,two weeks,four weeks and eight weeks after the surgery respectively.The tendency of the diameter change was analyzed.Results Twenty eight patients completed the whole monitor session,in which eleven were female.The average age of those patients was (53.68 ± 2.61) years old.Twelve of them were diabetic nephropathy.The diameters of all vessel were increased more rapidly at the first day than any other days after surgery(all P ＜ 0.01).The patients were divided into two groups depending on whether diabetic nephropathy.No significant difference was found between the two groups on the tendency of diameter change in cephalic vein and brachial artery (all P ＞ 0.05).However,the tendency of diameter change in radial artery and ulnar artery was statistically significant difference between the two groups (all P ＜ 0.05).Conclusions Cephalic vein,brachial artery,radial artery and ulnar artery are all apparently dilated on the first day after the surgery.The vascular dilation and diameter increasing become much slower after the period,the diameter tend to be stable.The primary diseases may affect the tendency of the diameter change in radial artery as well as ulnar artery.

BACKGROUND:In high-intensity exhaustive exercise process, the body must bear the exercise intensity decreasing splanchnic blood flow“ischemia”, at the same time, along with the movement of energy and material consumption, metabolite accumulation and oxidative stress in the body cause pathological damage, leading to a decline in exercise capacity. Thus, what is the impact on kidney filtration barrier? How to adapt to the change of renal tissue? Houttuynia cordata has the functions of heat clearing and detoxifying, dieresis for treating strangurtia, hemostatic, expel ing phlegm to arrest coughing and analgesia, if it has a protective effect on the renal injury caused by acute exhaustive exercise and its mechanism has not been reported in the literature. OBJECTIVE:To explore the effect of acute exhaustive exercise on kidney filtration barrier in rats and the intervention effect of Houttuynia cordata.METHODS:After resting and watching for 3 days, Sprague-Dawley rats received adaptive running for 15 minutes at a speed of 10 m/min on a 0° treadmil . A total of 24 rats, which can finish the running, were selected. They were divided into normal control group, exhaustive exercise group and dosed exhaustive exercise group according to the weight of layer (n=8). Rats in the exhaustive exercise group and dosed exhaustive exercise group on the 10° treadmil received once exhaustive exercise. Dosed exhaustive exercise group received intraperitoneal injection of sodium houttuyfonate 10 mL/kg at 30 minutes before exercises. The normal control group did not do any exercise. RESULTS AND CONCLUSION:Compared with the normal control group, serum urea nitrogen, serum creatinine, urine protein content, malondialdehyde concentration, renal cellapoptosis and apoptosis index were significantly increased, but nitric oxide content and nitric oxide synthase activity in the renal tissue were significantly deceased in the exhaustive exercise group (P<0.05 or 0.01). Glomerular filtration epithelial cells, the kidney filtration barrier of basement membrane and podocyte damage were obvious, showing abundant cellapoptosis, occasional y necrosis. Compared with the exhaustive exercise group, urine protein content, serum creatinine, malondialdehyde concentration, renal cellapoptosis and apoptosis index were significantly reduced, but nitric oxide content and nitric oxide synthase activity were significantly increased in the dosed exhaustive exercise group (P<0.05). No obvious pathological changes were detected, but apoptosis was visible. These findings confirmed that houttuynine made a reduction in renal cellinjury induced by exhaustive exercise and possibly significantly reduced apoptosis, increased nitric oxide synthase content, decreased malonaldehyde, and apparently increased superoxide dismutase activity, and final y protected injured renal tissue induced by exhaustive exercise.