Enhanced endoplasmic reticulum RyR1 receptor phosphorylation leads to diaphragmatic dysfunction in septic rats / 南方医科大学学报

OBJECTIVE@#To explore the role of endoplasmic reticulum ryanodine receptor 1 (RyR1) expression and phosphorylation in sepsis- induced diaphragm dysfunction.@*METHODS@#Thirty SPF male SD rats were randomized equally into 5 groups, including a sham-operated group, 3 sepsis model groups observed at 6, 12, or 24 h following cecal ligation and perforation (CLP; CLP-6h, CLP-12h, and CLP-24h groups, respectively), and a CLP-24h group with a single intraperitoneal injection of KN- 93 immediately after the operation (CLP-24h+KN-93 group). At the indicated time points, diaphragm samples were collected for measurement of compound muscle action potential (CMAP), fatigue index of the isolated diaphragm and fitted frequencycontraction curves. The protein expression levels of CaMK Ⅱ, RyR1 and P-RyR1 in the diaphragm were detected using Western blotting.@*RESULTS@#In the rat models of sepsis, the amplitude of diaphragm CMAP decreased and its duration increased with time following CLP, and the changes were the most obvious at 24 h and significantly attenuated by KN-93 treatment (P < 0.05). The diaphragm fatigue index increased progressively following CLP (P < 0.05) irrespective of KN- 93 treatment (P>0.05). The frequency-contraction curve of the diaphragm muscle decreased progressively following CLP, and was significantly lower in CLP-24 h group than in CLP-24 h+KN-93 group (P < 0.05). Compared with that in the sham-operated group, RyR1 expression level in the diaphragm was significantly lowered at 24 h (P < 0.05) but not at 6 or 12 following CLP, irrespective of KN-93 treatment; The expression level of P-RyR1 increased gradually with time after CLP, and was significantly lowered by KN-93 treatment at 24 h following CLP (P < 0.05). The expression level of CaMKⅡ increased significantly at 24 h following CLP, and was obviously lowered by KN-93 treatment (P < 0.05).@*CONCLUSION@#Sepsis causes diaphragmatic dysfunction by enhancing CaMK Ⅱ expression and RyR1 receptor phosphorylation in the endoplasmic reticulum of the diaphragm.

Relationship between intestinal microbiota heterogeneity and acquired myasthenia in septic mice / 中华麻醉学杂志

Objective:To evaluate the relationship between intestinal microbiota heterogeneity and acquired myasthenia in septic mice.Methods:Eighty SPF healthy male C57BL/6 mice, weighing 18-20 g, aged 6-8 weeks, were included. Forty mice were selected to prepare a sepsis model by intraperitoneal injection of lipopolysaccharide (LPS) 10 mg/kg. Sepsis-sensitive mice (state of dying or even death within 24 h after developing the model) and sepsis-resistant mice (survival for 7 days and recovery) were screened. The feces from donor mice were collected to make fecal bacteria fluid. Forty mice were selected and divided into 4 groups ( n=10 each) by the random number table method: control group (C group), antibiotic group (ABX group), resistant group (Res group), and sensitive group (Sen group). Group C received no treatment. In ABX group, compound antibiotics were given by intragastric gavage once a day for 5 consecutive days. In Res group, the fecal solution from sepsis-resistant mice 150 μl was given by gavage once a day for 3 consecutive days starting from 5 days after gavage administration of compound antibiotics, and then LPS 15 mg/kg was intraperitoneally injected. In Sen group, the fecal solution from sepsis-sensitive mice was given by gavage for 3 days (using the same method as previously described in Res group) starting from 5 days after gavage administration of compound antibiotics, and then LPS 15 mg/kg was intraperitoneally injected. The severity of sepsis was assessed and scored at 24 h after developing the model. The four limb grip strength was measured after fecal bacteria transplantation and at 24 h after developing the sepsis model. The Compound Muscle Action Potential (CMAP) of the gastrocnemius was measured at 24 h after developing the sepsis model. Then blood samples were collected for determination of the levels of interleukin-1 (IL-1), IL-6 and tumor necrosis factor α (TNF-α) in serum by enzyme-linked immunosorbent assay. The tissues of anterior tibial muscle and gastrocnemius muscle were obtained for determination of the expression of muscle-specific ring finger protein 1 (MuRF-1) and muscle atrophy box F protein (MAFbx) by Western blot. The diameter and cross-sectional area of gastrocnemius muscle fibers were measured after HE staining. The feces of mice were collected at 3 days after fecal bacteria transplantation, and DNA was extracted from mouse fecal samples for 16S rDNA sequencing and untargeted metabolomics analysis. Results:Compared with C group, the score for severity of sepsis was significantly increased, the four limb grip strength was decreased after developing the sepsis model, the serum IL-6 concentration was increased, and the diameter and cross-sectional area of gastnemius muscle fibers were decreased ( P<0.05), and no statistically significant changes were found in the other parameters in Res group, and no statistically significant changes were found in the indexes mentioned above in ABX group ( P>0.05). Compared with C group and Res group, the score for severity of sepsis was significantly increased, the four limb grip strength was decreased, the latency of CMAP was prolonged, and the amplitude of CMAP was decreased, the serum concentrations of IL-1, IL-6 and TNF-α were increased, the diameter and cross-sectional area of gastrocnemius muscle fibers were decreased, and the expression of MuRF-1 and MAFbx in anterior tibial muscle was up-regulated after developing the sepsis model in Sen group ( P<0.05). 16S rDNA sequencing of intestinal flora: Compared with Sen group, no significant change was found in Chao1 index, Good-coverage index and Simpson index in Res group ( P>0.05), Shannon index was increased ( P<0.05), and no significant change was found in β diversity in Res group ( P>0.05). LDA analysis showed that f_Akkermarisiaceae, o_Verrucomicrobiales, g_Akmansia, c_Verrucomicrobiae and p_Verrucomicrobiota were significantly enriched in Sen group, and g_Enterococcus and f_Enterococcaceae were significantly enriched in Res group ( P<0.05). Nontargeted metabolic analysis: The metabolite profiles in Res group and Sen group suggested that the model was well developed (R2Y>Q2Y). Compared with Sen group, 90 metabolites was significantly up-regulated and 88 down-regulated, significantly up-regulated metabolic substances including vitamin K1, gamma-tocopherol, and taurine in Res group ( P<0.05). The differential metabolite KEGG enrichment pathway included 2-oxocarboxylic acid metabolism and ubiquinone and other terpenoid-quinone biosynthesis ( P<0.05). Conclusions:Intestinal flora heterogeneity may be involved in the pathogenesis of acquired myasthenia in septic mice.

The association between indoleamine 2, 3-dioxygenase activity and clinical prognosis in patients with early stage non-small cell lung cancer after stereotactic radiotherapy / 中华放射医学与防护杂志

Objective:To explore the changes in activity of indoleamine 2, 3-dioxygenase(IDO)before and after stereotactic body radiation therapy (SBRT) in early stage non-small cell lung cancer(NSCLC) and its relationship with survival.Methods:The retrospective study was to assess IDO activity by serum kynurenine (Kyn) and Kyn to tryptophan (Trp) ratio before and after SBRT of early stage NSCLC.Thirty early stage NSCLC patients who received SBRT in the Cancer Hospital of the University of Chinese Academy of Sciences were enrolled, from December 2014 to July 2017. High-performance liquid chromatography and mass spectrometry were used to detect serum Kyn and Trp before and after SBRT. The ratios of Kyn after SBRT to Kyn before SBRT were divided into high and low group according to the median value. Similarly, the ratios of Kyn to Trp after SBRT were divided into high and low group.The correlation between overall survival (OS), progression-free survival (PFS) and IDO activity was evaluated. The factors influencing prognosis were analyzed.Results:In all patients, lower ratio of Kyn after SBRT to Kyn before SBRT significantly was correlated with better PFS (median PFS: not reached vs. 26.8 months, HR=0.31, 95% CI =0.11-0.90, P<0.05). The lower Kyn∶Trp ratio after SBRT had longer OS (median OS: not reached vs. 36.5 months, HR=0.27, 95% CI =0.079-0.95, P<0.05). In multivariate analysis, smoking <30 packs/year, higher BED, and lower ratio of Kyn after SBRT to Kyn before SBRT were associated with longer PFS. Lower Kyn∶Trp ratio after SBRT and higher BED were associated with longer OS. Conclusions:SBRT could alter IDO-mediated antitumor immune activity. IDO is a potentially valuable biomarker for monitoring immune status and predicting survival in early NSCLC patients after SBRT.

Influence of endothelin A receptor antagonist pretreatment on renal function after cardiopulmonary bypass in beagles / 重庆医学

Objective To observe the influence of endothelin A receptor antagonist pretreatment on renal function after cardiopulmonary bypass (CPB) in beagles.Methods A total of 18 male beagles were selected and allocated to 3 groups (n =6) by adopting the random number table method:sham operation group (Sham group),CPB group and endothelin A receptor antagonist (ETA) group.Sitaxsentan 0.7 mg/kg in the ETA group was infused by continuous pumping for 30 min at 1 h prior to CPB.The body temperature,mean arterial pressure (MAP) and arterial gas were collected.The concentrations of serum creatinine (Scr) and blood urea nitrogen (BUN) were detected.The renal tubular necrosis score was evaluated,and the expression levels of phosphorylated Akt (p-Akt),phosphorylated eNOS (p-eNOS) were also detected.Results Serum SCr and BUN levels at 2 h after CPB in the CPB and ETA group were significantly higher than those before operation (P＜0.05),and the ETA group was obviously lower than the CPB group (P＜0.05);the renal tubular necrosis score in the ETA group was obviously lower than that in the CPB group (P＜0.05).Expressions of p-Akt,p-eNOS in the ETA group were significantly higher than those in the CPB group(P＜0.05).Conclusion CPB might contribute to acute kidney injury,the endothelin A receptor antagonist pretreatment might alleviate acute kidney injury after CPB.

Inhibitory effect of a Chinese medicine Xiaoaiping combined with cisplatin on the proliferation, invasion and apoptosis in ovarian cancer HO-8910 PM cells in vitro and in vivo / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the effect of a Chinese medicine, Xiaoaiping, in combination with cisplatin on the proliferation, invasion and apoptosis in human ovarian cancer HO-8910PM cells in vitro and vivo.</p><p><b>METHODS</b>CCK-8 assay was used to detect the inhibitoty effect of Xiaoaiping alone or in combination with cisplatin on the proliferation of human ovarian cancer HO-8910PM cells. Flow cytometry was used to detect the apoptosis and cell cycle distribution. Transwell migration test was used to assay the effect of drugs on the cell invasive capability. Changes of the tumor volume in nude mice were observed to evaluate the antitumor effects in vivo.</p><p><b>RESULTS</b>CCK-8 assay Xiaoaiping alone or combined with cisplatin could inhibit the proliferation of HO-8910PM cells with a dose-dependent manner. The inhibition rates of Xiaoaiping combined with cisplatin were (53.4±3.0)%, significantly increased than those with single drug (P<0.05). Flow cytometry showed that G0/G1 fraction was increased respectively from (64.2±1.6)% to (74.1±1.6)% and (68.6±1.6)%. The percentages of apoptotic cells were increased from (2.2±1.6)% to (16.1±1.6)%, (35.6±1.6)% after treated with Xiaoaiping, Cisplatin and combination drugs (P<0.05 for all). Transwell chamber with matrigel assay showed that number of cells penetrating through membrane in HO-8910PM cells was (89.2±20.7)/HPF in the drug combination group, significantly less than that in the control group(187.2±24.6)/HPF, Xiaoaiping(141.8±13.7 )/HPF or cisplatin group (155.8±19.4)/HPF (P<0.01 for all). The inhibition rate of drug combination group in the nude mouse transplanted tumors, compared with that of single Xiaoaiping and cisplatin group, was increased significantly (59.0% vs. 23.4% and 34.2%), (P<0.01 for both).</p><p><b>CONCLUSION</b>The results of our in vitro and vivo experiments indicate that Xiaoaiping can inhibit cell proliferation, increase G0/G1 arrest, promote apoptosis, inhibit cell migration of human ovarian cancer HO-8910 PM cells, and can synergistically enhance the antitumor activity of cisplatine.</p>

The effects of a rotating magnetic field in the prevention and treatment of irradiation-induced esophagitis / 中华物理医学与康复杂志

Objective To observe the effect of a rotating magnetic field in preventing and treating irradiation-induced esophagitis in rats. Methods Forty female Wistar rats were randomized into 5 groups:a non-irradiated control group,an irradiation group,an amifostine treatment group ( amifostine group ),a 90 min magnetic field treatment group (90 min magnetic group) and a 120 min magnetic field treatment group ( 120 min magnetic group),with 8 rats in each group.The esophaguses of all rats except those in the control group were exposed to a single irradiation with 6 MV X-rays from a linear accelerator at a dosage of 43 Gy.Four rats in each group were randomly chosen to be observed 1 and 2 weeks after the irradiation.Blood cytokines were detected in their arterial blood.Any pathological changes of the esophagus were observed with HE staining under a light microscope at the same time. Results Irradiation-induced esophagitis was observed in the irradiation group 7 days after irradiation,with obvious exfoliation and necrosis of the esophagal epithelium mucosae.The submucosa were hyperaemic and dropsical with abundant inflammatory cell infiltration.The pathological changes of the esophagus were similar at 7 and 14 days after irradiation.However,the irradiation-induced esophagitis of rats in the amifostine group,the 90 min magnetic group and the 120 min magnetic group were relatively slighter and the blood leucocytes and neutrophis in those 3 groups were significantly lower than those in the irradiation group,while a tendency toward repair of the mucosa of the esophagus was detected.Serum TNF-α,IL-1 and IL-6 in the 90 min magnetic group and 120 min magnetic group were significantly lower than those in the irradiation group. Conclusions Both a rotating magnetic field and amifostine can help prevent and treat irradiation-induced esophagitis.Their therapeutic efficacy is similar.Exposure to a rotating magnetic field could inhibit the expression of inflammatory factors,and thus lessen the inflammatory reaction of acute irradiation-induced esophagitis.

Expression and clinical significance of guanylyl cyclase C mRNA in peripheral blood of the patients with colorectal cancer / 国际外科学杂志

Objective To investigate the expression of guanylyl eyelasec(GC-C)mRNA in pefipheral blood of colorectal cancer patients and discuss its clinical significance. Methods To detect the GC-C mRNA in peripheral blood of colorectal cancer patients by reverse transcriptase PCR and analysis their clinical and pathological indexes. Results The positive rate of GC-C mRNA in 52 colorectal cancer patients was 50%(26/52),while to patients with extra-intestinal cancer,the positive rate was 0(0/8).There were significant difierences of positive rate of GC-C mRNA associated with tumor infiltration(P＜0.05)and tumor embolus in vessel(P＜0.05).While to CEA level in peripheral blood(P＞0.05),but there were no statistical difference.The positive rate of micrometastasis in coloreetal cancer patients might increase using the GC-C mRNA combined with CEA level in peripheral bood.The positive rate of GC-C mRNA in colorectal cancer patients with Dukes A、B、C and D stages were 0(0/3),47.06%(8/17),52.38%(11/21),63.63%(7/11),The expression of GC-C mRNA increased with the clinical stage chang,there was no statistical difference.Conclusions GC-C expresses selectively in peripheral blood of colorectal cancer patients.Itis a valuable new tumor marker in micrometastasis diagnosis and post-operation follow-up of colorectal cancer,and it can also be used as the location determination of metastasis intestinal tumor.

Rotating magnetic fields and granisetron treatment for preventing nausea and vomiting induced by chemo-therapy / 中华物理医学与康复杂志

Objective To observe the efficacy of a rotating magnetic field and granisetron hydrochloride in preventing nausea and vomiting caused by a eisplatin regimen, and any side effects. Methods Sixty-eight patients receiving cisplatin regimen chemotherapy were randomly assigned to two groups: a magnetic treatment group and a drug treatment group. The patients in the two groups were exposed to a rotating magnetic field or received granisetron hydrochloride, respectively. The effects of the treatments were observed. Results Both treatments could effectively prevent and treat the vomiting caused by chemotherapy. The rate of response to the rotating magnetic field was 88.2% and to the drug 91.2%. However, tardive vomiting was significantly better controlled in the rotating magnetic field group. The incidence of side effects in the magnetic field group was 20.6% , and in the drug treatment group it was 45.6%. Conclusion The efficacy of a rotating magnetic field and granisetron in treating acute vomiting were simi- lar. The rotating magnetic field was more effective in preventing tardive vomiting and had fewer side effects. Magnetic therapy should be more generally applied in clinical practice.

Detection of CK19 mRNA and CK20 mRNA in peripheral blood cells of patients with early cervicalcancer and its clinical significance / 中国癌症杂志

Purpose:To study the expression of CK19 and CK2 0mRNA in peripheral blood patients with FRGO Stage ⅠA to ⅡA cervical carcinoma and to investigate its clinical significance. Methods:Using the reverse transcription polymerase chain reacti on(RT-PCR),CK19 and CK20mRNA expression was examined in peripheral blood fro m 250 patients with early cervical carcinoma before operation,50 Patients with benign gynecological tumors and 18 healthy volunteers. In 250 patients,possible correlations between clinical pathological factors were analyzed. Results:The positive expression rates of CK19 and CK20mRNA were 36% and 24% in 250 cervical carcinomas respectively,in comparison with 3.0% an d 0% with benign gynecological tumors and all subjects in healthy volunteers wer e negative; The expression of CK19 and CK20 mRNA were significantly correlated w ith lymph vascular space involvement,but was not associated with prognostic fac tors including stage,differentiation,pathological types ,lymph node metastasis ,bully tumor size . In patients with CK19 mRNA(+)/CK20 mRNA(+),the rate of lymp h node metastasis and vascular space involvement and recurrence outside the pelv is was significantly higher than that of patients with CK1R mNA9(-)/CK20 mRNA(- )(P

Proliferation and Immunophenotypic Analysis of CD3AK Cells Derived from Lymph Node / 中国肿瘤生物治疗杂志

Objective: To study the propagation and phenotypes changes of killer cell (CD3AK cell activated by CD3 mAb in vitro. Methods: Lymph nodes taken from lung cancer patient is dissociated into single cell suspension by mechanical method and cultured in culture medium added CD3 mAb and a little dose IL-2. We analyze cell immunophenotype by flow cytometry and proliferation by trypan blue exclusion test per 2 days. Results: Immunophenotypic analysis showed that CD3AK expressing CD3, CD8, CD56, CD25 increased, and reached a peak value which is 2.33 times than before culturing in the 8 th day. Conclusion: CD3 mAb added to the culture medium can obviously activate CD3AK cell and stimulate proliferation and keep its killer activity.