Objective To investigate correlation between the expression level of multiple cytokine levels in neonatal umbilical cord plasma and early-onset sepsis for screening out the cytokines with good diagnostic value for early-onset neonatal sepsis(EONS).Methods Full-term neonates and preterm neonates(Gestational age ≥ 32 weeks)of 310 cases between September 2021 and June 2023 were selected as study subjects.According to clinical signs,laboratory results and blood culture,these subjects were divided into 3 groups:control group without sepsis,EONS blood culture positive group and EONS blood culture negative group.Umbilical cord blood plasma of all subjects was collected within 72 hours after birth.The expression levels of cytokines IL-2,IL-4,IL-6,IL-9,IL-10,IL-21,IFN-γ and TNF-α were determined,and cytokines with high expression levels(high correlation)were screened out.Receiver operating characteristic(ROC)curve was used to analyze the specificity and sensitivity of the selected cytokines in the diagnosis of neonatal early-onset sepsis.Results Among the 8 cytokines mentioned above,the concentrations of IL-6,IL-9 and IL-21 in cord blood plasma of neonatal early-onset sepsis positive blood culture patients(392.6±258.7pg/ml,11.9±7.5pg/ml,29.1±16.8 pg/ml)and negative blood culture patients(353.8±244.5pg/ml,10.4±6.3pg/ml,27.7±19.2pg/ml)were higher than those of the control group(34.9±25.1pg/ml,5.9±4.5pg/ml,10.8±10.1 pg/ml),with significant differences(t=23.961,20.732;15.174,17.824;22.466,21.193,all P＜0.01),and the increase of IL-6 concentration was the most obvious.ROC curve analysis(the cut-off values of IL-6,IL-9 and IL-21:123.0 pg/ml,3.60 pg/ml,6.00 pg/ml,respectively)showed that the areas under the ROC curve for IL-6,IL-9 and IL-21 alone detection were 0.876(95％CI:0.786～0.955),0.782(95％CI:0.667～0.875)and 0.825(95％CI:0.737～0.913),respectively.The area under the ROC curve for the combined detection of IL-6,IL-9 and IL-21 was 0.930(95％CI:0.875～0.997).The combined detection of IL-6,IL-9 and IL-21 improved the specificity and sensitivity of the test than IL-6,IL-9 and IL-21 alone detection,and the differences were statistically significant(Z=2.137,2.391,2.257,all P＜0.05).There was no significant difference in cytokine expression between positive blood culture and negative blood culture neonates with early-onset sepsis(t=0.276～3.377,all P＞0.05).Conclusion The cytokines expression of IL-6,IL-9 and IL-21 in neonatal umbilical cord plasma of neonatal early-onset sepsis were increased.Combined detection of IL-6,IL-9 and IL-21 has good diagnostic value for early-onset neonatal sepsis.

Methods:Whole blood and serum samples were collected from 86 patients with HBV-ACLF and 42 patients with chronic hepatitis B (CHB). Serum alanine aminotransferase (ALT), total bilirubin (TBil), creatinine (Cr), albumin (ALB) cholinesterase (CHE) and international standardized ratio (INR) were detected by routine method . The CXCL10 mRNA level of peripheral blood mononuclear cells was detected by fluorescence quantitative PCR. The genotype and allele of rs56061981 were detected by direct DNA sequencing. The genotype and allele frequencies between the ACLF and CHB groups were compared. The levels of ALT, TBil, INR, ALB, CHE, model for end-stage liver disease (MELD) and CXCL10 mRNA were compared between the two groups according to whether the T allele was carried.Results:There was no significant difference in sex, age, alcohol history, smoking history, HBV genotype, HBeAg status and HBV-DNA level between CHB group and HBV-ACLF group (P > 0.05). The frequency of CT+ TT genotype and the frequency of T allele at the rs56061981 locus of the CXCL10 gene in group ACLF were significantly higher than that in the CHB group ( χ2=4.83, P=0.03 and χ2=4.95, P=0.03). The difference is significant Genotype CT+ TT is a risk factor for CHB to develop into HBV-ACLF ( OR=2.897, 95% CI: 1.09~7.68). Allele T is a risk factor for CHB to develop into HBV-ACLF ( OR=2.746, 95% CI: 1.10~6.89). The plasma INR and MELD scores of ACLF individuals carrying T alleles were significantly higher than those carrying C alleles (t=2.63, P=0.013 and t=2.7, P=0.011). The difference is significant. Albumin (ALB) and cholinesterase (CHE) was significantly lower than those carrying C alleles (t=2.67, P=0.01 and t=3.545, P=0.001). The difference is significant. But there was no significant difference in serum TBL and ALT between the two groups. Conclusions:The polymorphism of CXCL10 gene rs56061981 is significantly correlated with the susceptibility and severity of ACLF. Allele T may be a susceptible gene of ACLF, and it can also be used as a predictor of the severity of the disease.

Objective: To predict the tumor neoantigen peptides in hepatocellular carcinoma (HCC), and examine their specific immune effects against the tumor cells without injury to normal cells. Methods: The data of whole-genome sequencing and exome sequencing of HCC tumor and matched non-tumor liver tissues were analyzed to confirm the HCC-associated somatic mutations. Based on the HLA phenotype of the patients, we used NetMHC software to predict the neoantigen epitopes with high binding affinity to their MHC-I molecules. The predicted peptides with mutation sites included were synthesized. GPL10687 platform was applied to examine the gene expression difference between tumor and normal tissues of the selected genes in GSE25097, one of the GEO databases. The quantitative real-time PCR (qRT-qPCR) and immunohistochemistry were used to confirm the expressions in tumors and normal tissues of the selected genes. By using the predicted peptides, we induced the generation of antigen-specific CD8⁺ cytotoxic T lymphocytes (CTLs) and examined their specific effects against tumor cells. Results: The mutation frequency of TP53 (tumor protein p53) was 40%, and LAMA3 (Laminin Subunit Alpha 3) was 8% in the analyzed HCC tissues. In GSE25097 database, TP53 and LAMA3 mRNA levels in tumors were 1.57±0.02 and 1.37±0.10, which were significantly increased than those in matched no-tumor tissue (0.54±0.01 and 0.36±0.01, P<0.05). The differences of expression levels of TP53 and LAMA3 in tumor and no-tumor tissues were validated by using qRT-qPCR and immunohistochemistry in 10 HCC tissues. The mRNA levels of TP53 and LAMA3 in tumors were 0.24±0.03 and 0.13±0.06, which were significantly elevated than those in matched no-tumor tissue (0.11±0.01 and 0.01±0.01, P<0.05). Among the Chinese population, HLA-A2 and HLA-A11 and HLA-A24 accounted for 70%, representing the major MHC-I molecules. The CTLs induced by predicted peptides showed cytotoxicity to the targets pulsed with mutated peptide, with no effect on the target pulsed with normal peptide and on normal cells. Conclusions TP53 and LAMA3 existed relative higher mutation frequency in HCC, and expressed higher in tumor tissues. The induced CTLs by predicted peptides derived from mutation-associated protein could specific kill the target cells without injury to normal cells.

Objective To predict the tumor neoantigen peptides in hepatocellular carcinoma (HCC), and examine their specific immune effects against the tumor cells without injury to normal cells. Methods The data of whole?genome sequencing and exome sequencing of HCC tumor and matched non?tumor liver tissues were analyzed to confirm the HCC?associated somatic mutations. Based on the HLA phenotype of the patients, we used NetMHC software to predict the neoantigen epitopes with high binding affinity to their MHC?I molecules. The predicted peptides with mutation sites included were synthesized. GPL10687 platform was applied to examine the gene expression difference between tumor and normal tissues of the selected genes in GSE25097, one of the GEO databases.The quantitative real?time PCR (qRT?qPCR) and immunohistochemistry were used to confirm the expressions in tumors and normal tissues of the selected genes. By using the predicted peptides, we induced the generation of antigen?specific CD8+ cytotoxic T lymphocytes ( CTLs ) and examined their specific effects against tumor cells. Results The mutation frequency of TP53 (tumor protein p53) was 40%, and LAMA3 (Laminin Subunit Alpha 3) was 8% in the analyzed HCC tissues. In GSE25097 database, TP53 and LAMA3 mRNA levels in tumors were 1.57±0.02 and 1.37±0.10, which were significantly increased than those in matched no?tumor tissue (0.54±0.01 and 0.36±0.01, P＜0.05). The differences of expression levels of TP53 and LAMA3 in tumor and no?tumor tissues were validated by using qRT?qPCR and immunohistochemistry in 10 HCC tissues. The mRNA levels of TP53 and LAMA3 in tumors were 0.24±0.03 and 0.13±0.06, which were significantly elevated than those in matched no?tumor tissue (0.11±0.01 and 0.01±0.01, P＜0.05). Among the Chinese population, HLA?A2 and HLA?A11 and HLA?A24 accounted for 70%, representing the major MHC?I molecules. The CTLs induced by predicted peptides showed cytotoxicity to the targets pulsed with mutated peptide, with no effect on the target pulsed with normal peptide and on normal cells. Conclusions TP53 and LAMA3 existed relative higher mutation frequency in HCC, and expressed higher in tumor tissues. The induced CTLs by predicted peptides derived from mutation?associated protein could specific kill the target cells without injury to normal cells.

Objective To predict the tumor neoantigen peptides in hepatocellular carcinoma (HCC), and examine their specific immune effects against the tumor cells without injury to normal cells. Methods The data of whole?genome sequencing and exome sequencing of HCC tumor and matched non?tumor liver tissues were analyzed to confirm the HCC?associated somatic mutations. Based on the HLA phenotype of the patients, we used NetMHC software to predict the neoantigen epitopes with high binding affinity to their MHC?I molecules. The predicted peptides with mutation sites included were synthesized. GPL10687 platform was applied to examine the gene expression difference between tumor and normal tissues of the selected genes in GSE25097, one of the GEO databases.The quantitative real?time PCR (qRT?qPCR) and immunohistochemistry were used to confirm the expressions in tumors and normal tissues of the selected genes. By using the predicted peptides, we induced the generation of antigen?specific CD8+ cytotoxic T lymphocytes ( CTLs ) and examined their specific effects against tumor cells. Results The mutation frequency of TP53 (tumor protein p53) was 40%, and LAMA3 (Laminin Subunit Alpha 3) was 8% in the analyzed HCC tissues. In GSE25097 database, TP53 and LAMA3 mRNA levels in tumors were 1.57±0.02 and 1.37±0.10, which were significantly increased than those in matched no?tumor tissue (0.54±0.01 and 0.36±0.01, P＜0.05). The differences of expression levels of TP53 and LAMA3 in tumor and no?tumor tissues were validated by using qRT?qPCR and immunohistochemistry in 10 HCC tissues. The mRNA levels of TP53 and LAMA3 in tumors were 0.24±0.03 and 0.13±0.06, which were significantly elevated than those in matched no?tumor tissue (0.11±0.01 and 0.01±0.01, P＜0.05). Among the Chinese population, HLA?A2 and HLA?A11 and HLA?A24 accounted for 70%, representing the major MHC?I molecules. The CTLs induced by predicted peptides showed cytotoxicity to the targets pulsed with mutated peptide, with no effect on the target pulsed with normal peptide and on normal cells. Conclusions TP53 and LAMA3 existed relative higher mutation frequency in HCC, and expressed higher in tumor tissues. The induced CTLs by predicted peptides derived from mutation?associated protein could specific kill the target cells without injury to normal cells.

Objective To explore the role of magnetic resonance spectroscopy in evaluating the severity and prognosis of patients with brain stem injury. Methods A total of 45 patients with brain injury (the study group)and 10 healthy people(the control group)were retrospectively analyzed in Affiliated Anqing Hospital,Anhui Medical University during March 2015 to October 2016 using magnetic resonance spectroscopy.The NAA/Cr,NAA/Cho and Cho/Cr value in brain stem were analyzed. The clinical severity score (GCS) and Glasgow outcome scale (GOS)were also analyzed in our study. Results Compared with the normal group,patients in the study group had lower NAA level, NAA/Cho value but higher Cho/Cr value (P < 0.05). The severer the injury, the more obvious in NAA peak decrease. Cho peak increased and NAA/Cho ratio decreased. NAA/Cho, NAA/Cr and Cho/Cr values in the group with good prognosis were similar to those in the normal control group, but significantly different to the group with poor prognosis (P < 0.05). NAA/Cr, NAA/Cho and Cho/Cr values correlates with GOS in patients with brain stem injury. Conclusions Magnetic resonance spectroscopy can be used to measure the metabolic status of patients with brain stem injury,which is of great significance in the evaluation the severity of brain stem injury and prognosis.

Objective To observe the expression of aquaporin 4 (AQP-4) in contused brain tissue and its relationship with brain edema following brain trauma.Methods A retrospective case control analysis was made on 42 patients with severe brain trauma admitted from January 2015 to March 2016.There were 23 males and 19 females,aged from 23 to 62 years [(35.5 ± 5.6) years].Glasgow coma score (GCS) was 3-5 points in 7 patients,6-8 points in 23 and 9-10 points in 12.Brain tissue removed from the area 1 cm near the contusion during the cranial surgery were allocated to study group (n =42),while brain tissue removed far from the contusion after internal decompression were used as control (n =8).Ultrastructure of brain tissues was observed under electron microscope.Water content of brain tissue was measured by dry-wet weight method and expression of AQP-4 was measured by immunohistochemical method at postinjury hours ＜ 6,6-12,12-24,24-72,72-96 and ＞ 96.Results Morphology and structure of brain tissue in control group were normal.Whereas in study group,the intracellular and interstitial edema were obvious and morphological structure were damaged.Water content and AQP-4 expression in control group showed no obvious increase after operation(73.55 ±0.10,0.193 ±0.016).Water content in study group increased significantly compared to control group and reached the peak value (81.28 ± 0.56) at postinjury 24-72 hours (P ＜ 0.01).AQP-4 expression in study group increased at postinjury 6 hours (0.242 ±0.023) and reached a peak at postinjury 24-72 hours (0.338 ± 0.013),with significant difference compared to control group (P ＜ 0.05).Correlation analysis showed change of brain water content was positively correlated with expression level of AQP-4 (r =0.931,P ＜ 0.01).Conclusion Expression of AQP-4 in the injured area of brain trauma is significantly increased along with the increase of water content,suggests that the upregulation of AQP-4 plays an important role in traumatic brain edema.

Objective: To investigate the efficacy and safety of the new investigational drug pegylated interferon α-2b (Peg-IFN-α-2b) (Y shape, 40 kD) injection (180 µg/week) combined with ribavirin in the treatment of patients with genotype 1/6 chronic hepatitis C (CHC), with standard-dose Peg-IFN-α-2a combined with ribavirin as a positive control. Methods: A multicenter, randomized, open-label, and positive-controlled phase III clinical trial was performed. Eligible patients with genotype 1/6 CHC were screened out and randomly divided into Peg-IFN-α-2b(Y shape, 40kD) group and Peg-IFN-α-2a group at a ratio of 2:1. The patients in both groups were given oral ribavirin for 48 weeks in addition and then followed up for 24 weeks after drug withdrawal. Abbott Real Time HCV Genotype II was used to determine HCV genotype, and Cobas TaqMan quantitative real-time PCR was used to measure HCV RNA level at 0, 4, 12, 24, 48, and 72 weeks. Adverse events were recorded in detail. The primary efficacy endpoint was sustained virological response (SVR), and a non-inferiority test was also performed. Results: A total of 561 patients with genotype 1/6 CHC were enrolled, among whom 529 received treatment; 90.9% of these patients had genotype 1 CHC. The data of the full analysis set showed that SVR rate was 69.80% (95% CI 65.00%-74.60%) in the trial group and 74.16% (95% CI 67.73%-80.59%) in the control group (P = 0.297 0). The data of the per protocol set (PPS) showed that SVR rate was 80.63% (95% CI 76.04%-85.23%) in the trial group and 81.33% (95% CI 75.10%-87.57%) in the control group (P = 0.849 8), and the 95% CI of rate difference conformed to the non-inferiority standard. The analysis of the PPS population showed that of all subjects, 47.9% achieved rapid virologic response, with a positive predictive value of 93.8%. The incidence rate of adverse events was 96.30% in the trial group and 94.94% in the control group, and the incidence rate of serious adverse events was 5.13% in the trail group and 5.06% in the control group. Conclusion In the regimen of Peg-IFN-α combined with ribavirin for the treatment of genotype 1/6 CHC, the new investigational drug Peg-IFN-α-2b(Y shape, 40 kD) has comparable clinical effect and safety to the control drug Peg-IFN-α-2a.

Objective: To investigate the value of jugulo-omohyoid lymph nodes (JOHLN) in predicting occult lateral cervical lymph node metastasis in patients with papillary thyroid carcinoma (PTC). Methods: The clinicopathological data of 136 out of 2 100 PTC patients, who had a high risk of lateral neck lymph node metastasis and treated by us from January 2010 to December 2015, were retrospectively analyzed. Super selective neck dissection (SSND, level Ⅲ and Ⅳ)was performed and JOHLNs were sent for frozen section in all the 136 cases. The clinicopathological data was analyzed and the significance of JOHLN in predicting lateral cervical LNM was calculated using the SPSS software package. Results: Of the 136 patients, total thyroidectomy was performed in 76 cases (55.9%) and unilateral lobectomy plus isthmus was performed in the other 60 cases (44.1%). SSND was performed in 72 patients (52.9%), level Ⅱ-Ⅳ dissection in 15 (11.0%), and level Ⅱ-Ⅴ dissection in 49 (36.0%). According to the pathological results, 38 patients were pN0(27.9%), 18 (13.2%) were pN1a and 80 (58.8%) were pN1b. The lymph node metastasis(LNM) rates at level Ⅱ-Ⅵ were 19.9%, 43.4%, 42.6%, 2.9%, and 59.6%, respectively. The sensitivity, specificity and accuracy of JOHLN in predicting lateral neck metastasis were 58.8%, 62.9%, and 76.7%, respectively. The rates for predicting level Ⅱ metastasis were 81.5%, 43.2%, and 59.4%, respectively. None of the patients died in the follow-up. Only 1 recurrence was found in level Ⅱ and regional control was achieved after level Ⅱ and Ⅴ dissection. Conclusions JOHLN has a high accuracy for predicting lateral cervical lymph node metastasis and high sensitivity for level Ⅱ metastasis. For patients with high risk of lateral cervival metastasis, super-selective neck dissection including level Ⅲ and Ⅳ can confirm the stage and reduce the risk of reoperation. Dissection for level Ⅱ, Ⅲ, and Ⅳ is recommended.

Objective To investigate the surgical treatment of growth hormone (GH) pituitary tumors combined with cardiomyopathy patients and its clinical effect.Methods A total of 15 patients with GH pituitary tumors combined with cardiomyopathy were enrolled and underwent surgical treatment from February 2007 to May 2015 in our hospital.Pituitary endocrinology,brain magnetic resonance imaging,echocardiography diagram were inspected before and after surgery.Serum GH level,left ventricular diastolic diameter (LVIDd),thickness of the interventricular septum (IVST),left ventricular posterior wall thickness (LVPWT),left ventricular ejection fraction (EF),and mitral valve flow spectrum ratio (E/A) were detected by noninvasive methods and compared before and after treatment.Results Through the pituitary adenomas via nasal-sphenoid sinus surgery,total removal of the tumors were noted in 9 patients,subtotal resection in 4 patients,and partial resection in two patients.GH level,LVIDd,IVST,GH,LVPWT,E/A and EF after resection of pituitary tumor were significantly improved as compared with those before resection (P＜0.05).In the long-term follow-up,the cure rate was 35.68％,and the remission rate was 76.36％.Conclusion Surgical resection of pituitary tumor with good control of growth hormone levels can significantly improve the cardiac structure and function,thereby,GH pituitary tumor patients with cardiomyopathy can obtain satisfactory therapeutic effect.