ObjectiveTo investigate the influence of heat shock protein A2 (HSPA2) on the biological behavior of pancreatic adenocarcinoma cells and its mechanism. MethodsThe expressions of HSPA2 were determined in the human pancreatic adenocarcinoma cell lines (PANC-1, BxPC-3, and AsPC-1) using the Western blot. Subsequently, the cells with the lowest and highest HSPA2 expressions among these three lines were selected for conducting overexpression and knockdown experiments targeting HSPA2, respectively. The cellular proliferation, cell clonogenesis, migration, and invasion capabilities were assessed using MTT, clonogenic assay, and Transwell assay, respectively. Additionally, the impact of HSPA2 on the expression of key markers of epithelial-mesenchymal transition (EMT) was examined using the Western blot. The potential target molecules of HSPA2 were identified through immunoprecipitation assay and mass spectrometry. The rescue experiments further explored the regulatory relation between the HSPA2 and its target molecules. The influence of HSPA2 on pancreatic adenocarcinoma growth was investigated through establishment of xenograft tumor model in nude mice. ResultsThe HSPA2 exhibited the lowest expression in the PANC-1 cells and the highest expression in the AsPC-1 cells among the three cell lines. Subsequent functional studies demonstrated that the overexpression of HSPA2 in the PANC-1 cells markedly promoted proliferation, cell clonogenesis, migration, and invasion, while the knockdown of HSPA2 expression in the AsPC-1 cells markedly inhibited these processes. The Western blot analysis further showed that the HSPA2 overexpression downregulated E-cadherin expression and upregulated N-cadherin and Vimentin expressiones, whereas the HSPA2 knockdown produced opposite effects. The rescue experiments indicated that the HSPA2 promoted the EMT in pancreatic adenocarcinoma cells by upregulating Yes associated protein (YAP). The subcutaneous xenograft tumor experiments in the nude mice showed that the HSPA2 knockdown inhibited tumor growth. ConclusionThe results of this study suggest that HSPA2 promotes EMT via upregulating YAP, which facilitates proliferation, migration, and invasion of pancreatic adenocarcinoma cells.
ObjectiveTo evaluate the predictive value of mini-fluid challenge for volume responsiveness in patients under shock.MethodsSixty patients diagnosed as shock were included in the study. A 50 mL infusion of physiological saline over 10 seconds and a further 450 mL over 15 minutes were conducted through the central venous catheter. Cardiac output (CO), global end-diastolic volume index (GEDVI), central venous pressure (CVP) and extravascular pulmonary water index (EVLWI) were monitored by the pulse indicator continuous cardiac output monitoring. If the increase of CO after 500 mL volume expansion (ΔCO500) ≥10%, the patient was considered to be with volume responsiveness. The relevance between ΔCO50 and ΔCO500 was analyzed, and the sensitivity and specificity of the ΔCO50 were analyzed by receiver operating characteristic (ROC) curve.ResultsAfter 50 mL volume injection, the heart rate and systolic blood pressure of the two groups did not change obviously. The CVP of non-responders changed slightly higher than that of responders, but neither of them had obviously difference (P>0.05). The CO of responders had increased significantly (P<0.05) which was in accord with that after a further 450 mL volume injection. GEDVI and EVLWI did not change significantly (P>0.05). ΔCO50 and ΔCO500 were strongly correlated (r=0.706, 95%CI 0.677 - 0.891, P>0.05). The area under ROC curve for ΔCO50 was 0.814 (95%CI 0.707 - 0.922).ConclusionThe volume responsiveness of patients under shock can be predicted by mini-fluid challenge study which is related to normal volume expansion and it does not increase the risk of pulmonary edema.
Objective To detect the expression of heat shock protein 47 mRNA in pathological scar tissue by using real-time fluorescent quantitative reversetranscription-polymerase chain reaction (RT-PCR). Methods The tissues of normal skin(n=6), hypertrophic scar(n=6) and keloid(n=6) were adopted, which were diagnosised by Pathology Department. Based on fluorescent TaqMan methodology, the real-time fluorescent quantitative RT-PCR were adopted to detect the expression ofheat shock protein 47 mRNA. Results Compared with normal skin tissue（0.019±0.021）×105, the expressions of heat shock protein47 cDNA of hypertrophic scar tissue（1.233±1.039）×105 and keloid tissue（1.222±0.707）×105 were higher， being significant differences（Plt;0.05）. Conclusion A fluorescent quantitative method was successfully applied to detecting the expression of heat shock protein 47 mRNA. Heat shock protein 47 may play an important role in promoting the formation of pathological scar tissue.
ObjectiveTo observe the influence of heat shock protein 27 (HSP27) sensibilization to retinal ganglion cells (RGC) apoptosis of rats. MethodsThirty-five female Wistar rats were randomly divided into HSP27 sensibilization group (15 rats), borate buffer solution (BBS) control group (15 rats) and normal group (5 rats). The rats in HSP27 sensibilization group were received hypodermic injection in rear limb with 100 μg HSP27 and complete freund adjuvant, intraperitoneal injection with 1 μg pertussis toxin. The BBS control group received the same volume of BBS at the same site. The normal group received no intervention. The intraocular pressure was measured 3 days before injection and 1, 2, 4, 6, 8 weeks after injection. Four, 6 and 8 weeks after injection, the retinal frozen sections was made to observe RGC apoptosis by terminal-deoxynucleoitidyl transferase mediated nick end labeling. The anti-HSP27 level in serum and cerebrospinal fluid were detected by enzyme linked immunosorbent assay. ResultsThere was no obvious change of intraocular pressure in rats in 3 groups before injection (P>0.05). RGC apoptosis was observed in HSP27 sensibilization group 4 weeks after injection, and increased significantly at 6 weeks after injection. There was no RGC apoptosis in BBS control group and normal group. The level of anti-HSP27 in serum and cerebrospinal fluid of HSP27 sensibilization group occurred at 4 and 6 weeks after injection respectively, decreased with prolongation of injection time. Compared with BBS control group and normal group, the RGC apoptosis rate, anti-HSP27 level in serum and cerebrospinal fluid of HSP27 sensibilization group were significantly increased (P<0.05). There was no significant difference of the RGC apoptosis rate, anti-HSP27 level in serum and cerebrospinal fluid between BBS control group and normal group (P>0.05). ConclusionsHSP27 sensibilization could promote the RGC apoptosis. The variation trend of anti-HSP27 level in cerebrospinal fluid is consistent with the RGC apoptosis.
Electron spin resonance (ESR) spectroscopy and spin trapping agent PBN were applied to measure directly the changes of oxygen free redicals (OFR) in gastric mucosa of rats with portal hypertension (PHT) injured by shockreperfusion, and treated with superoxide dismutase (SOD), radix salviae miltiorrhizae (RSM), with concomitant monitoring activity of SOD and pathology of gastric mucosa. Results showed that the amount of OFR increased markedly in gastric mucosa of PHT rats during the shock-reperfusion. The pathological changes were in accordance with alteration of the amount of OFR and the activity of SOD. Gastric mucosa in PHT was more susceptible to shock-reperfusion insult than normal controls. The anti-oxidant SOD, RSM used at early stage exerted mild gastric mucosal insult through different mechanisms.
Objective To investigate the expressions of heat shock protein 27 (HSP27), Bcl-2, and Bax proteins of the nerve cells after spinal cord ischemia/reperfusion injury (SCII) in rats and their relationship. Methods Seventy adult male Sprague Dawley rats (weighing, 200-220 g) were randomly divided into the sham operated group (sham group, n=35) and the SCII group (n=35). Only the left renal artery was exposed with no occlusion of the abdominal aorta in the rats of sham group. The left renal artery was exposed with occlusion of the abdominal aorta for 20 minutes in the rats of SCII group. At 4, 8, and 12 hours and at 1, 2, 3, and 5 days, reperfusion treatment was performed in 5 rats respectively, and then the spinal cord tissue was harvested to detect the expressions of HSP27, Bcl-2, and Bax protein of the nerve cells by using immunohistochemistry staining. Results The HSP27 began to express at 4 hours, reached the peak at 3 days, and decreased at 5 days in SCII group; significant differences were found between at 3 and 5 days and at the other time points (P lt; 0.05). The Bcl-2 expression increased at 4 hours, reached the peak at 1 day and maintained a high level at 2 days, and then gradually decreased; significant differences were found between at 1 and 2 days and at the other time points (P lt; 0.05). The Bax expression reached the peak at 12 hours and 3 days, and decreased at 5 days; significant differences were found between at 12 hours and 3 days and at the other time points (P lt; 0.05). A little expression of each protein was observed in sham group at different time points; the expressions of HSP27, Bcl-2, and Bax proteins in SCII group were significantly higher than those in sham group at different time points (P lt; 0.05). Conclusion There may be the time window of self repair after SCII. High expression of HSP27 has an obvious protective effect on the SCII in rat, by promoting the expression of the anti-apoptotic protein Bcl-2 and reducing the expression of the pro-apoptotic protein Bax so as to inhibit spinal cord cell apoptosis.
Objective To evaluate the effects of intensive care unit (ICU)-acquired hypernatremia (IAH) on the outcome of septic shock patients. Methods This retrospective study analyzed 116 septic shock patients admitted to the ICU of the First Affiliated Hospital of Soochow University from August 2018 to December 2022. Patients were divided into two groups: IAH group and normonatremia group. χ2 test, t test and the Mann-Whitney U test of the non-parametric test were used to compare the differences in clinical data between the two groups. Independent risk factors for IAH were identified by unconditioned Logistic regression analysis, and receiver operating characteristic (ROC) curves were constructed to determine their role in predicting IAH. The Kaplan-Meier curve was used to evaluate the effects of IAH and its duration on 28-day survival. Results Renal insufficiency, K+ concentration, body temperature max, mechanical ventilation, chronic critical illness, rapid recovery, sepsis-associated encephalopathy, persistent inflammation, immunosuppression and catabolism syndrome, and the length of stay in ICU had significant differences (P<0.05). Multivariate logistic regression analysis showed: total urine volume in the previous 3 days [odds ratio (OR) 1.09, 95% confidence interval (CI) 1.01 - 1.16, P=0.019] and sodium content in enteral nutrition preparations (670 mg) (OR 6.00, 95%CI 1.61 - 22.42, P=0.006) were independent risk factors for IAH. In addition, the area under the ROC curve of total urine output in the first 3 days was 0.800 (95%CI 0.678 - 0.922, P=0.001). Finally, the duration of IAH was significantly correlated with 28-day survival rate (P=0.020). Conclusions IAH is a common and serious complication in septic shock, and is the main cause of poor prognosis. Sodium status may act as an ideal screening tool for patients with septic shock.