Objective:To evaluate the effect of sleep deprivation on the expression of sirtuin 6 (SIRT6) in the cerebellum of immature mice.Methods:Fifty SPF healthy male C57BL/6 mice, aged 4 weeks, weighing 14-16 g, were divided into 2 groups ( n=25 each) using a random number table method: control group (Con group) and sleep deprivation group (SD group). The chronic sleep deprivation model was prepared by using the multi-platform water environment method, with 20 h of sleep deprivation per day for 10 consecutive days. After sleep deprivation, a balance beam experiment was performed to test the balance and coordination ability of mice. The mice were sacrificed after anesthesia and cerebellar lobular IV-VI (4-6 cb) tissues were taken for microscopic examination of the ultrastructure (with a transmission electron microscope) and for determination of the dendritic spine density of cerebellar 4-6cb Purkinje neurons (by Golgi staining), co-expression of SIRT6 and Calbindin D-28k (CbD-28k) and expression of glucose transporter Glut3 of cerebellar 4-6cb (by immunofluorescence staining). Results:Compared with group Con, the duration of passage through the balance beam was significantly prolonged, and the number of posterior foot slips was increased, the synaptic gap of cerebellar 4-6cb neurons was increased, the thickness of postsynaptic density was increased, the density of dendritic spines of Purkinje cells and the number of positive cells co-expressing SIRT6 and CbD-28k were decreased, and the expression of Glut3 was down-regulated in group SD ( P<0.05). Conclusions:The mechanism by which sleep deprivation decreases the abilities of balance and coordination is related to down-regulating SIRT6 expression in cerebellar Purkinje cells and decreasing neuronal glucose metabolism, thus damaging the synaptic plasticity of cerebellum in immature mice.

Objective:To detect gene mutations in a pedigree with dominant dystrophic epidermolysis bullosa (DDEB) .Methods:A 20-year-old male proband presented with repeated blisters, ulceration, pigmentation, scars on the limbs, and deformation of the nails/toenails after birth. There were 5 patients in the 3-generation family, and they all presented with typical skin lesions. Peripheral blood samples were obtained from 14 members of the pedigree (including the 5 patients) and 100 unrelated healthy controls. Whole-exome sequencing was performed in the proband to identify relevant mutation sites, which were then confirmed in the family by Sanger sequencing.Results:Genetic testing indicated that the proband and the other 4 patients all carried a missense mutation (c.7885G>A) in exon 107 of the COL7A1 gene, resulting in the substitution of glycine by arginine at amino acid position 2629 (p.G2629R). The mutation was identified neither in the 9 healthy relatives nor in the 100 unrelated healthy controls. The mutation co-segregated with DDEB in the family, and was not included in databases such as Pubmed, HGMD or ClinVar, suggesting it was a novel missense mutation. The amino acid encoded by this mutation may alter the structure of type Ⅶ collagen, thereby affecting its function.Conclusion:A novel missense mutation was identified in exon 107 of the COL7A1 gene in the family with DDEB, expanding the spectrum of mutations in the COL7A1 gene.

Objective:To investigate the clinical efficacy of modified biliary-intestinal anasto-mosis by pancreaticoduodenectomy and influencing factors of postoperative biliary leakage.Methods:The propensity score matching and retrospective cohort study was conducted. The clinicopatholo-gical data of 165 patients with benign and malignant diseases around the ampullary who underwent pancreaticoduodenectomy in the Affiliated Hospital of Inner Mongolia Medical University from June 2014 to October 2020 were collected. There were 92 males and 73 females, aged (59±10)years. Of the 165 patients, 44 patients undergoing modified biliary-intestinal anastomosis within pancreatico-duodenectomy were divided into the modified group, and 121 patients undergoing traditional biliary-intestinal anastomosis within pancreaticoduodenectomy were divided into the traditional group. Observation indicators: (1) propensity score matching and comparison of general data of patients between the two groups after matching; (2) intraoperative and postoperative situations; (3) analysis of influencing factors of biliary leakage after pancreaticoduodenectomy. Propensity score matching was done by the 1:1 nearest neighbor matching method, with the caliper setting as 0.05. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the 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. Univariate analysis was conducted using the corresponding statistical methods based on data type. All indicators in univariate analysis were included in multivariate analysis. Multivariate analysis was conducted using the Logistic regression model. Results:(1) Propensity score matching and comparison of general data of patients between the two groups after matching. Of the 165 patients, 72 cases were successfully matched, including 36 cases in the modified group and 36 cases in the traditional group, respectively. The elimination of jaundice, preoperative reduction of jaundice and hypertension confounding bias ensured comparability between the two groups after propensity score matching. (2) Intraoperative and postoperative situations. All patients in the two groups underwent surgery successfully. The operation time, postoperative pathological type (lower bile duct cancer, pancreatic head cancer, pancreatic cystic tumor, chronic pancreatitis, duodenal cancer), time of no drainage fluid in the drainage tube around biliary-intestinal anastomosis were 371(270,545)minutes, 6, 12, 1, 2, 15, (12±7)days in patients of the modified group, versus 314(182,483) minutes, 13, 14, 1, 4, 4, (16±8)days in patients of the traditional group, showing significant differences in the above indicators between the two groups ( Z=-3.54, χ2=10.01, t=-2.34, P<0.05). Cases with postoperative grade A biliary leakage was 0 in patients of the modified group, versus 6 in patients of the traditional group, showing a significant difference between the two groups ( P<0.05). Cases with postoperative grade B biliary leakage, cases with postoperative grade B pancreatic fistula, cases with postoperative bleeding, cases with abdominal infection, cases with incision infection, cases with delayed gastric emptying, cases undergoing unplanned readmission were 1, 0, 1, 4, 1, 5, 1 in patients of the modified group, versus 0, 1, 2, 5, 2, 5, 2 in patients of the traditional group, showing no significant difference in the above indicators between the two groups ( P>0.05). Cases with postoperative grade A pancreatic fistula, cases with overall complications, cases with Clavien-Dindo grade Ⅰ-Ⅱ complications, cases with Clavien-Dindo grade Ⅲ-Ⅳ complications were 6, 12, 6, 6 in patients of the modified group, versus 7, 14, 8, 6 in patients of the traditional group, showing no significant difference in the above indicators between the two groups ( χ2=0.09, 0.24, 0.36, 0.00, P>0.05). None of patient in the two groups had postoperative grade C biliary leakage and postoperative grade C pancreatic fistula. (3) Analysis of influencing factors of biliary leakage after pancreaticoduodenectomy. Results of multivariate analysis showed that preoperative reduction of jaundice and traditional biliary-intestinal anastomosis were independent risk factors for biliary leakage after pancreaticoduodenectomy ( odds ratio=11.37, 12.27, 95% confidence interval as 1.76-73.35, 1.14-131.23, P<0.05). Conclusions:Compared with traditional biliary-intestinal anastomosis, modified biliary-intestinal anastomosis within pancreaticoduodenectomy is safe and feasible. Preoperative reduction of jaundice and traditional biliary-intestinal anastomosis are independent risk factors for biliary leakage after pancreaticoduodenectomy.

Objective:To evaluate the prognostic accuracy of the sequential organ failure assessment (SOFA), quick sequential organ failure assessment (qSOFA) and systemic inflammatory response syndrome (SIRS) criteria in predicting the mortality in patients with infection or suspected infection by using network Meta-analysis.Methods:Five databases including Wanfang Data, China National Knowledge Infrastructure (CNKI), China Science and Technology Journal Database (VIP), PubMed, Web of Science were searched from February 23, 2016 to September 5, 2020 to identify the relevant literatures comparing the prognostic accuracy of two or more scores for mortality in patients with infection or suspected infection. The literatures screening, data extraction and the quality assessment of the included studies were all conducted independently by two reviewers. Stata 14.0 software was used to test the heterogeneity between the original studies of pairwise comparison of each of the three scoring systems. Ring inconsistency test was used to judge the consistency between direct comparison and indirect comparison. Then network Meta-analysis was performed and the results were ranked. The predictive ability of the three scoring systems was evaluated by surface under cumulative ranking curve (SUCRA). A "comparison-correction" funnel plot was drawn to assess whether there was publication bias in the included studies.Results:A total of 38 studies were enrolled, the overall quality was high. Network meta-analysis showed that SOFA had a great prognostic performance in predicting mortality for patients with infection or suspected infection, which was followed by qSOFA [mean difference ( MD) = 0.07, 95% confidence interval (95% CI) was 0.05-0.09] and SIRS scores ( MD = 0.16, 95% CI was 0.14-0.18), and the qSOFA score was better than SIRS score ( MD = 0.09, 95% CI was 0.07-0.11). In the order of predicting the death risk of patients with infection or suspected infection, SOFA score had higher predictive value, followed by qSOFA score, and SIRS score was the lowest, with SUCRA values of 1.0, 0.5 and 0, respectively. Funnel plot showed that all the studies were distributed on both sides of the midline, but the distribution was not symmetrical, suggesting that there was a high possibility of publication bias and small sample effect. Conclusions:SOFA score had the best prognostic performance in predicting mortality of patients with infection or suspected infection as compared with qSOFA score and SIRS score. However, the funnel plot showed that included literatures may exist small sample effects or publication bias. So the final results should be validated by more prospective studies with multicenters and large samples.

Objective:To investigate changes of nicastrin (NCSTN) downstream molecules in signaling pathways related to cell proliferation and differentiation after silencing the expression of the NCSTN gene in the human immortalized keratinocyte cell line HaCaT.Methods:HaCaT cells were divided into 3 groups: interference group transfected with a specific small interfering RNA (siRNA) targeting NCSTN (NCSTN-siRNA) , negative control group transfected with a negative control siRNA, and blank control group transfected with the equal amount of transfection reagent. Real-time PCR and Western blot analysis were performed to measure the NCSTN mRNA and protein expression in groups, in order to verify the transfection efficiency. Differences in gene expression profiles in HaCaT cells were detected between the interference group and negative control group by using Agilent whole-genome microarray, and differentially expressed genes were identified based on a fold change ≥ 2.0 with a P value ≤ 0.05. Gene ontology (GO) enrichment analysis was employed to identify the roles of the differentially expressed genes, and then to screen out significantly differentially expressed genes associated with proliferation and differentiation of keratinocytes, some of which were verified by real-time PCR. Results:The interference group showed significantly decreased mRNA and protein expression of NCSTN (0.287 ± 0.090, 0.443 ± 0.085, respectively) compared with the negative control group (0.969 ± 0.127, 1.047 ± 0.114, respectively) and blank control group (1.000 ± 0.151, 1.000 ± 0.111, F = 30.787, 31.139, respectively, both P = 0.001) . Whole genome-expression analysis using an Agilent microarray platform revealed 605 downregulated genes and 444 upregulated genes in HaCaT cells in the interference group compared with the negative control group. GO analysis showed that differentially expressed genes were enriched into 4 biological processes, including epithelial development, epithelial cell differentiation, keratinocyte differentiation and keratinization. The significantly differentially expressed genes associated with proliferation and differentiation of keratinocytes, including the Sprouty-related protein with EVH1 domain 2, fibroblast growth factor 7, insulin-like growth factor-binding protein 5, Rho-associated coiled-coil kinase 2 and bone morphogenetic protein 6 genes, were verified by real-time PCR, and the verification results were consistent with the difference trend shown by the microarray results. Conclusion:The loss of NCSTN gene function may affect the normal proliferation and differentiation of keratinocytes by regulating the expression of its downstream molecules in signaling pathways associated with cell proliferation and differentiation.

Acne inversa is a chronic inflammatory skin disease of the folliculo-sebaceous-apocrine unit. Currently, genetic and immunological factors are hot topics in the study of its pathogenesis. Genetic factors are mainly related to γ-secretase mutations, and abnormal expression of the γ-secretase-Notch axis leads to increased keratinization of hair follicles and inflammation in some patients with haploinsufficiency of the γ-secretase gene. Mutations in the γ-secretase gene are not necessary for acne inversa, and the risk of Alzheimer′s disease in familial acne inversa patients still remains unclear. Some progress has been made in researches on the association of genotype with phenotype in acne inversa patients, but further studies with large sample size are needed for verification.

Objective To evaluate the clinical application of Catalyst system in patient positioning during breast cancer radiotherapy,and to analyze its correlation with age and body mass index (BMI).Methods Twenty-four patients with breast cancer who were admitted to our hospital from May to August,2016 were enrolled as subjects.For all patients,auxiliary positioning was made by the optical surface imaging system (CRad Catalyst) before each treatment.The kV-kV imaging was executed weekly to verify positioning.Age,BMI,and setup errors of the two systems in the anterior-posterior (AP),superior-inferior (SI),and left-fight (LR) directions were recorded and analyzed by independent samples t-test and Pearson correlation analysis.Results The C-Rad Catalyst system had a significantly larger setup error in the AP direction than the kV-kV imaging (0.22±0.17 vs.0.18±0.13 cm,P＜0.05).There were no significant differences in setup errors in the SI or LR direction between the two systems (0.23±0.18 vs.0.19±0.15 cm,P＞0.05;0.28±0.28 vs.0.20±0.15 cm,P＞ 0.05).Age and BMI of patients had significant impacts on the C-Rad Catalyst system but the kV-kV imaging (P＞0.05):there were significant differences in setup errors in the AP and SI directions between patients ≤44 years of age and those 45-59 years of age (all P＜0.05);there were significant differences in setup errors in the AP and LR directions between patients ≤44 years of age and those ≥60 years of age (all P＜0.05);there was a significant difference in setup error in the LR direction between patients 45-59 years of age and those ≥ 60 years of age (P＜0.05);there was a significant difference in setup error in the SI direction between patients with BMIs of＜ 25 and ≥ 25 kg/m2 (P＜ 0.05).For patients ≥ 60 years of age,setup error of the C-Rad Catalyst system in the SI direction was correlated with age (r=-0.496,P＜0.05).For patients with BMI of＜25 kg/m2,setup error of the C-Rad Catalyst system in the AP direction was correlated with BMI (r=-0.445,P＜ 0.05).For patients with a BMI of ≥ 25 kg/m2,setup error of the C-Rad Catalyst system in the SI direction was correlated with BMI (r=-0.252,P＜0.05).Conclusions There is significant difference in setup error in the AP direction between the C-Rad Catalyst system and the kV-kV imaging.Age and BMI have impacts on patient positioning by the C-Rad Catalyst system.

Objective To analyze γ?secretase gene mutations in a pedigree with acne inversa. Methods Clinical data were collected from a pedigree with acne inversa, which contained 30 members spanning 4 generations. Of these members, 12 were affected by acne inversa, and 9 of the affected members were alive. Peripheral blood DNA was obtained from the proband, his seven relatives (including 4 affected and 3 unaffected members), and 100 unrelatedhealthy human controls. PCR was performed to amplify all the coding exons and their flanking sequences of the NCSTN, PSEN1, PSENEN, Aph1 genes followed by DNA sequencing. Results A heterozygous insertion mutation (c.229_230insCACC)of the PSENEN gene, which led to translational frameshifting and resulted in dysfunciton of the PSENEN protein, was detected in all the 5 patients, but not in unaffected members or healthy controls. Conclusion There is a novel heterozygous insertion mutation c.229_230insCACC in the PSENEN gene, which may be the molecular basis of acne inversa in this family.

Objective To measure expressions of nicastrin and its downstream Notch-HES signaling pathwayassociated proteins in skin lesions of patients with acne inversa harbouring nicastrin gene mutations.Methods An immunohistochemical study was performed to measure the expressions of nicastrin and Notch-HES signaling pathwayassociated proteins in paraffin-embeded skin samples from lesions of 4 patients with acne inversa and confirmed nicastrin mutations and from normal skin of 6 human controls.Spearman correlation analysis was carried out to assess the relationship between the expressions of nicastrin and Notch-HES signaling pathway-associated proteins.Results In normal control skin samples,nicastrin was widely distributed in the full-thickness epidermis and skin appendages such as pilosebaceous units,apocrine glands and eccrine glands.However,the expressions of nicastrin and Notch-HES signaling pathway-associated proteins were markedly decreased in the epidermis and hair follicle infundibulum in lesions of patients harbouring nicastrin gene mutations compared with normal control skin.Furthermore,nicastrin expression was positively correlated with Notchl,Notch3 and HES-1 expressions (r =0.831,0.748 and 0.807,P ＜ 0.01,0.05 and 0.01 respectively),but not significantly correlated with Notch2 or HES-5 expressions (r =0.597,0.591 respectively,both P ＞0.05).Conclusion Nicastrin expression markedly decreases in lesions of patients with acne inversa harbouring nicastrin gene mutations,and is positively correlated with the expressions of several Notch-HES signaling pathway-associated proteins,suggesting that the decrease in nicastrin expression may take part in the pathogenesis of acne inversa by influencing the expression of the downstream Notch-HES signaling pathway.

OBJECTIVE: Lymphoepithelial carcinoma (LEC) of salivary glands is a rare malignant neoplasm. The purpose of this research was to investigate the clinicalpathologic features and treatment methods of this rare disease. METHOD: The clinical data and treatment outcomes of 17 patients from 2006 to 2012 were reviewed retrospectively. RESULT: Ten males and seven females with a ratio of 1. 43:1 were involved. The II, III, IV stage cases were 7 (41.2%), 4 (23.5%), 6 (35.3%), respectively. The average follow-up duration was 2.56 years, and 12 patients had no evidence of recurrence. Five patients had local recurrence and (or) distant metastases within three years after surgery, including 4 deaths. CONCLUSION LEC in salivary gland is a high grade malignant tumor in oral and maxillofacial region, occurring predominately in parotid gland and submandibular gland. To prevent distant metastasis, radical surgery combined with chemoradiotherapy should be adopted.
Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasms, Squamous Cell ; pathology ; secondary ; Parotid Neoplasms ; pathology ; Rare Diseases ; pathology ; Retrospective Studies ; Salivary Gland Neoplasms ; pathology ; Salivary Glands ; pathology ; Submandibular Gland Neoplasms ; pathology ; Treatment Outcome