AIM: To study the effect of L-arginine (L-Arg) on function and structure of mitochondria in ischemia-reperfusion (MRI) myocardial cells. METHODS: Thirty rabbits were randomly divided into three groups (n=10 in each), control group, MIR group and MIR+L-Arg group. The mitochondrial respiratory function, Ca~(2+) concentration ([Ca~(2+)]m), malondialdehyde (MDA) content and superoxide dismutase (SOD) activity were determined. Meanwhile, the contents of adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), total adenylic acid number (TAN) and energy charge (EC) in the myocardial tissue were respectively measured. Moreover, the ultrastructure changes in myocardial mitochondria were observed during MIR. RESULTS: The mitochondrial respiratory control rate (RCR), velocity 3 (V_3), SOD, surface density (Sv) and specific surface (?) in MIR+L-Arg group were higher than those in MIR group, velocity 4 (V_4), [Ca~(2+)]m, MDA, volume density (Vv), horizental diameter (Hd) were lower than those in MIR group. ATP, ADP, TAN and EC levels of myocardial tissue were higher than those in MIR group. There was no significant difference between MIR+L-Arg and control group in V_3, V_4, SOD, MDA, Vv, Sv, ?, Nv, Vd, AMP and TNA. CONCLUSION: It is suggested that L-Arg improves the function and structure of mitochondria in myocardial cells in the reperfusion injury after myocardial ischemia by decreasing oxygen free radical level and Ca~(2+) overload in the mitochondria. [

Objective To investigate the characteristics and risk factors of central lymph node metastasis in clinically node negative (cN0) papillary thyroid carcinoma (PTC) (T1 or T2 stage) coexisting with Hashimoto' s thyroiditis (HT).Methods A total of 398 patients undergoing thyroidectomy with central lymph node dissection were enrolled in the study.Patients were divided into the trial group (PTC with HT)and the control group (PTC without HT).The difference of the clinicopathological characteristics between the 2 groups and risk factors for central lymph node metastasis were analyzed.Results Among the total 398 patients,98 (24.6％)had coexistent HT.Central lymph node metastasis rate was similar in the 2 groups (40.8％ vs 41.3％).The number of dissected central lymph nodes was significantly more in the trial group than in the control group (4.9 vs 2.9,P＜0.01) while the number of metastatic lymph nodes had no statistical significance between the 2 groups (1.0 vs 1.0).Univariate analysis showed that tumor size＞1 cm was significantly associated with central lymph node metastasis in the trial group (P＜0.01).Male,＜45 years,tumor size＞1 cm,and tumor located in the middle/lower third of lobe were all significantly associated with central lymph node metastasis in the control group (P＜0.01).Multivariate analysis showed that tumor size＞1 cm was independent predictor for central lymph node metastasis in the trial group,while female,＜45 years,tumor size＞1 cm,and tumor located in the middle/lower third of lobe were all independent predictors for central lymph node metastasis in the control group.Conclusions The number of central lymph nodes was larger in cN0 PTC coexisting with HT patients than that in PTC patients,but there was no statistical difference in the number of metastatic lymph nodes between cN0 PTC with and without HT.Central lymph node dissection is recommended when tumor size 1 cm in cN0 PTC coexisting with HT patients.

Objective To investigate the effects of carcinoembryonic antigen (CEA), carbohy-drate antigen (CA19-9), carbohy-drate antigen (CA125), human epididymis protein 4 (HE4) And the combined detection of ovarian cancer risk prediction model (ROMA) in the diagnosis and significance of endometrial cancer.Methods From May 2014 to May 2010, 80 patients with endometrial cancer were treated.According to its clinical pathology is divided into observation 1 group and observation 2 group.Serum samples were collected and serum CEA, CA19-9, CA125 and HE4 levels were analyzed statistically and the ROMA values were calculated according to menopausal status.Results Compared with the control group, the levels of CEA, CA19-9, CA125 and HE4 in the serum of observation group patients with endometrial carcinoma were significantly higher than those in the control group, the ROMA value of the menopausal state was significantly higher, the difference was statistically significant (P＜0.05).And with the progression of endometrial cancer staging, compared with the observation 1 group, the levels of CA19-9, CA125 and HE4 in the serum of the observation 2 group were significantly higher, the ROMA values of the menopausal states were significantly increased, the difference was statistically significant (P＜0.05).The combined detection rate of CEA, CA19-9, CA125, HE4 and ROMA in serum of patients with endometrial carcinoma was significantly higher than that of each index (P＜0.05).Conclusion The abnormally elevated levels of CEA, CA19-9, CA125, HE4, and ROMA in the serum of patients have important significance and role in the clinical diagnosis of endometrial cancer and its clinical staging.

Porcine interferon-alpha (pIFN-alpha) fermentative production by recombinant Pichia pastoris was carried out in a 10-L bioreactor to study its metabolism changes and effects on fermentation under different inducing strategies, by analyzing the change patterns of the corresponding metabolism and energy regeneration. The results show that the specific activities of alcohol oxidase (AOX), formaldehyde dehydrogenase (FLD) and formate dehydrogenase (FDH) largely increased when reducing temperature from 30 degrees C to 20 degrees C under pure methanol induction, leading significant enhancements in methanol metabolism, formaldehyde dissimilatory energy metabolism and pIFN-alpha antiviral activity. The highest pIFN-alpha antiviral activity reached 1.4 x 10(6) IU/mL, which was about 10-folds of that obtained under 30 degrees C induction. Using methanol/sorbitol co-feeding strategy at 30 degrees C, the major energy metabolism energizing pIFN-alpha synthesis shifted from formaldehyde dissimilatory energy metabolism pathway to TCA cycle, formaldehyde dissimilatory pathway was weakened and accumulation of toxic intermediate metabolite-formaldehyde was relieved, and methanol flux distribution towards to pIFN-alpha synthesis was enhanced. Under this condition, the highest pIFN-alpha antiviral activity reached 1.8 x 10(7) IU/mL which was about 100-folds of that obtained under pure methanol induction at 30 degrees C. More important, enhanced pIFN-alpha production with methanol/sorbitol co-feeding strategy could be implemented under mild conditions, which greatly reduced the fermentation costs and improved the entire fermentation performance.
Animals ; Energy Metabolism ; Fermentation ; Interferon-alpha ; biosynthesis ; genetics ; Methanol ; pharmacology ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; Sorbitol ; pharmacology ; Swine

Objective:To investigate the expression of lymphoid enhancement factor-1 (LEF-1) in Wnt signaling pathway in pancreatic ductal adenocarcinoma (PDAC) and its significance.Methods:The relative expressions of LEF-1 mRNA in human PDAC cell line PANC-1 and normal pancreatic ductal epithelial cell line HPDE6 were detected by fluorescence quantitative PCR. A total of 45 pancreatic cancer tissue specimens and their corresponding paracancerous tissue specimens were collected from the Department of Hepatobiliary and Pancreatic Surgery, Affiliated Tumor Hospital of Xinjiang Medical University from June 2012 to December 2013. The expressions of LEF-1 in the cancer tissues and paracancerous tissues were detected by immunohistochemistry, and the relationships between LEF-1 expression and clinicopathological characteristics and prognosis of patients were analyzed.Results:Fluorescence quantitative PCR showed that the relative expression level of LEF-1 mRNA in PANC-1 cell line was significantly higher than that in HPDE-6 cell line (2.895±0.485 vs. 1.006±0.126, t=3.056, P<0.001). Immunohistochemical results showed that LEF-1 was highly expressed in 33 cases (73.3%) of cancer tissues, which was higher than that in 12 cases (26.7%) of adjacent tissues, and there was a statistically significant difference ( χ2=14.815, P<0.001). LEF-1 expression was correlated with preoperative carbohydrate antigen (CA) 19-9 level ( P<0.001) and local lymph node metastasis ( P=0.041). Survival analysis showed that the median overall survival (OS) was 22.0 months in patients with PDAC, 19.0 months in patients with high LEF-1 expression ( n=33), 31.0 months in patients with low LEF-1 expression ( n=12), and there was a statistically significant difference ( χ2=5.554, P=0.018). Univariate Cox regression analysis showed that age ( HR=1.962, 95% CI: 1.043-3.692, P=0.037), LEF-1 ( HR=2.253, 95% CI: 1.097-4.630, P=0.027), and CA19-9 ( HR=2.667, 95% CI: 1.258-5.656, P=0.011) were associated with OS. Multivariate Cox regression analysis showed that CA19-9 ( HR=6.431, 95% CI: 1.078-38.382, P=0.041), CA125 ( HR=0.151, 95% CI: 0.027-0.839, P=0.031), primary tumor size ( HR=8.364, 95% CI: 1.925-36.335, P=0.005), LEF-1 ( HR=2.281, 95% CI: 1.025-5.075, P=0.043) were independent risk factors affecting the prognosis of PDAC patients. Conclusion:LEF-1 expression is up-regulated in PDAC tissues, which is positively correlated with preoperative CA19-9 level and local lymph node metastasis, and is an independent prognostic factor in patients with PDAC.

Objective:To investigate the expression level of β-catenin and its relationship with clinicopathology and prognosis of patients with pancreatic ductal adenocarcinoma (PDAC).Methods:Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of β-catenin mRNA in primary pancreatic cancer cell line and pancreatic ductal epithelial cell line HPDE6-C7 of the healthy. The data of 45 patients with PDAC confirmed by pathology at Xinjiang Medical University Cancer Hospital from June 2012 to December 2013 were retrospectively analyzed. Immunohistochemical method was used to detect the expression level of β-catenin in cancer tissues and adjacent tissues, and the correlation of β-catenin with pathological characteristics of patients with PDAC was analyzed. Cox proportional hazard model was performed to make univariate and multivariate analysis on the influencing factors of overall survival (OS).Results:The relative expression of β-catenin mRNA in primary pancreatic cancer cells was higher than that in HPDE6-C7 cell line [(3.83±0.83) vs. (1.00±0.03)], and the difference was statistically significant ( t = 3.45, P = 0.003). The high expression rate of β-catenin protein in PDAC tissues was higher than that in para-cancer tissues [68.9% (31/45) vs. 28.9% (14/45)], and the difference was statistically significant ( χ2 = 7.50, P = 0.005). The high expression rate of β-catenin protein in PDAC patients with different tumor diameter and TNM staging had statistically significant differences ( P = 0.026, P = 0.036). The median OS time of 45 patients was 22.5 months, and that of high expression of β-catenin protein group in 31 patients was 19 months, that of low expression of β-catenin group in 14 patients was 29 months, and the difference was statistically significant ( P = 0.009). Univariate Cox analysis showed that preoperative carbohydrate antigen199 (CA199) level, tumor diameter, tumor differentiation degree and the expression level of β-catenin protein were influencing factors of OS of patients with PDAC. Multivariate Cox analysis showed that preoperative CA199 ( OR = 9.883, 95% CI 2.815-34.689, P < 0.001), tumor diameter ( OR = 6.117, 95% CI 1.578-24.179, P = 0.009), tumor differentiation degree ( OR = 3.834, 95% CI 1.158-12.697, P = 0.028), the expression level of β-catenin protein ( OR = 0.139, 95% CI 0.045-0.430, P = 0.001) were independent affecting factors of OS of patients with PADC. Conclusions:β-catenin is abnormally highly expressed in PDAC which is correlated with the disease progression of patients and may be a new indicator and therapeutic target of prognosis for PDAC patients.

Objective:To investigate the clinical effect of respiratory intervention training in feeding of preterm infants, so as to provide reference for feeding interventions in preterm infants.Methods:From March 2021 to October 2021, the 80 preterm infants hospitalized in the Neonatal Intensive Care Unit of Xuzhou Central Hospital were selected and divided into intervention group (40 cases) and control group (40 cases) according to the random table method in a prospective study. The common nursing was given in the control group, the respiratory intervention training was given on this basis in the intervention group, once a day for two weeks. The intervention effect was evaluated by the time of indwelling gastric tube, hospital stay, feeding reaction, feeding related complications and satisfaction rate of catch-up growth. The oral feeding ability of preterm infants was evaluated by the non-nutritive sucking (NNS) score and preterm infants oral feeding readiness assessment scale (PIOFRAS) score.Results:The time of indwelling gastric tube and hospital stay was (15.3 ± 8.7), (28.9 ± 9.8) d in the intervention group and (21.7 ± 9.8), (34.6 ± 12.2) d in the control group, the difference was significant ( t=3.12, 2.32, both P<0.05). After intervention, the differences of respiratory number, heart rate and pulse oxygen saturation were (6.5 ± 1.3) beats/min, (11.2 ± 1.5) beats/min, 0.048 ± 0.015 in the intervention group and (11.2 ± 1.2) beats/min, (16.5 ± 1.3) beats/min, 0.082 ± 0.018 in the control group, the differences were significant ( t=16.39, 16.40, 9.35, all P<0.05). The incidence of feeding-related complications was 25.0% (10/40) in the intervention group and 55.0% (22/40) in the control group, the difference was significant ( χ2=7.50, P<0.05). The satisfaction rate of catch-up growth was 77.5%(31/40) in the intervention group and 40.0%(16/40) in the control group, the difference was significant ( χ2=12.78, P<0.05). The scores of NNS and PIOFRAS at 7 d, 10 d and 14 d after intervention were (13.73 ± 2.24), (29.98 ± 6.67), (49.08 ± 16.37) points and (15.28 ± 1.41), (25.08 ± 3.10), (31.03 ± 3.00) points in the intervention group and (10.30 ± 2.18), (15.68 ± 2.42), (28.60 ± 3.61) points and (12.60 ± 1.22), (17.15 ± 1.51), (23.75 ± 3.87) points in the control group, the differences were significant ( t values were -14.53--6.94, all P<0.05). Conclusions:The respiratory intervention training can effectively shorten the transition time from tube feeding to complete oral feeding in preterm infants, reduce feeding reaction and the incidence of feeding-related complications, and promote the coordination of feeding behavior, so as to improve oral feeding ability and catch-up growth of preterm infants.

Objective:To explore the anxiety level, influencing factors among surrogate decision-makers of patients with acute ischemic stroke during thrombolysis decision-making, and their correlation with decision-making duration.Methods:Acute ischemic stroke patients and their surrogate decision-makers who visited the Emergency Department of the First Affiliated Hospital of Zhengzhou University from September 2019 to December 2021 were selected as the research subjects.Sociodemographic data and disease related data of patients and surrogate decision-makers were collected.Surrogate decision-makers were evaluated with the state-trait anxiety inventory, decision participation expectation scale, Wake Forest physician trust scale, and perceived social support scale.SPSS 26.0 software was used for data processing.Pearson correlation analysis, Spearman correlation analysis and ridge regression analysis were used for statistical analysis.Results:The score of state anxiety of decision-makers was (49.47±9.04), and 18.2% (70/383) of decision-makers had a decision duration exceeding 15 minutes.The score of state anxiety of decision-makers was positively correlated with decision duration ( r=0.189, P<0.001). The influencing factors of state anxiety level of decision-makers included sociodemographic factors (age of decision-makers and patients, relationship between payers and patients, whether decision-makers bear the current medical expenses, type of medical insurance for patients), psychological factors (trust level in physicians, perceived social support), factors related to patient disease (numbers of stroke relapses, National Institutes of Health stroke scale scores for patients), characteristics of the decision-making process (whether patients participate in the decision-making process, and the role of decision-makers in the decision-making process) (all P<0.05). Conclusion:Most surrogate decision-makers experience anxiety.Medical staff should pay attention to the emotions of decision-makers and adopt appropriate communication skills when communicating with informed consent for thrombolysis, alleviate the anxiety of surrogate decision-makers, so as so reduce the decision-making duration.

Objective: To analyze the etiology and epidemiological characteristics of influenza in Wenzhou from 2009 to 2014, so as to provide the scientific basis for control and prevention of influenza. Methods: Throat swab specimens of influenza like illness (ILI) were collected from national influenza surveillance sentinel hospitals for nucleic acid detection with real-time PCR and virus isolation, culture and sequencing, and the results were analyzed with statistical methods. Results: During the 8 years, a total of 10 577 089 cases from outpatient and emergency department were monitored in sentinel hospitals. There were 337 896 ILI cases with an average ILI treatment rate of 3.19%. A total of 4 046 ILI samples were detected in children, 511 were positive for influenza, the positive rate was 12.63%. Among the detected influenza types, type B had the highest proportion, followed by H3N2. Among the 6 age groups, the number of flu patients was the highest in 0-3 years old group, the positive rate in 10-12 years old group was the highest (35.03%). There were 28 and 45 amino acid sequence mutations of HA fragment in influenza A and B, respectively, which included multiple mutation of 391 and 145 amino acids. The phylogenetic analysis showed that the strains of type B were different in different years, and Yamagata evolved into Y1 and Y2 two branches. Conclusions The prevalence peaks of influenza in children occurred in winter and spring in Wenzhou city, accompanied by small peaks in summer. Three subtypes of serotypes B, H3N2 and A(H1N1) dominated alternatively in Wenzhou during the 8 years. We should focus on strengthening the prevention and control of influenza in preschool children and primary and secondary school students.

Objective: To investigate the inhibitory effect of bortezomib (PS-341) on enterovirus replication. Methods: The methyl thiazolyl tetrazolium (MTT) assay was used to value cell viability in response to PS-341 treatment. The protein and viral gene mRNAs were measured by real-time quantitative PCR (qRT-PCR). Results: Our result show that after enterovirus (EV)-D68 or coxsackievirus B3 (CV-B3) infected cells were treated with PS-341, compared with the control group, the inhibition rate of the intracellular viral RNA reached 50%~70% or 60%~90%. PS-341 was added after RD cells were infectd with EV-D68, the intracellular virus titer was down-regulated by 90.23% and 83.40% in the supernatant, the intracellular virus titer was down-regulated by 93% and 90% in the supernatant and in RD cells. PS-341 had no effect on virus adsorption and importing. The cells were treated with PS-341 and apoptosis-inhibiting agent Ac-YVAD-CHO, the viral RNA replication inhibition rate reached 10%-30%, and the expression of viral protein was increased, which indicated that the inhibitory effect of PS-341 on viral replication was attenuated. Conclusions According to the result of the study, PS-341 could reduce apoptosis by regulating the proteasome pathway, inhibiting the gene replication and assemble, without effect on virus adsorption, entry and release. In addition, PS-341 also inhibited the replication of CV-B3 in cells, which suggest that PS-341 has a broad spectrum anti-EVs effects.