Humans ; Sodium-Calcium Exchanger ; Carrier Proteins ; Pain ; Calcium/metabolism*

OBJECTIVES: To analyze the clinical and genetic characteristics of children with autosomal dominant neurodevelopmental disorders caused by kinesin family member 1A (KIF1A) gene variation. METHODS: Clinical and genetic testing data of 6 children with KIF1A gene de novo heterozygous variation diagnosed in Shanghai Children's Medical Center Affiliated to Shanghai Jiao Tong University School of Medicine from the year 2018 to 2020 were retrospectively analyzed. Pathogenic variants were identified based on whole exome sequencing, and verified by Sanger sequencing. Moreover, the effect of variants on three-dimensional structure and stability of protein was analyzed by bioinformatics. RESULTS: Among 6 patients there were 4 males and 2 females, and the age of consultation varied from 7 months to 18 years. All cases had varying degrees of motor developmental delay since childhood, and 4 of them had gait abnormalities or fell easily. In addition, 2 children were accompanied by delayed mental development, epilepsy and abnormal eye development. Genetic tests showed that all 6 cases had heterozygous de novo variations of KIF1A gene, including 4 missense mutations c.296C>T (p.T99M), c.761G>A (p.R254Q), c.326G>T (p.G109V), c.745C>G (p.L249V) and one splicing mutation c.798+1G>A, among which the last three variants have not been previously reported. Bioinformatics analysis showed that G109V and L249V may impair their interaction with the neighboring amino acid residues, thereby impacting protein function and reducing protein stability, and were assessed as "likely pathogenic". Meanwhile, c.798+1G>A may damage an alpha helix in the motor domain of the KIF1A protein, and was assessed as "likely pathogenic". CONCLUSIONS KIF1A-associated neurological diseases are clinically heterogeneous, with motor developmental delay and abnormal gait often being the most common clinical features. The clinical symptoms in T99M carriers are more severe, while those in R254Q carriers are relatively mild.
Male ; Female ; Humans ; Child ; Retrospective Studies ; China ; Mutation ; Epilepsy/genetics* ; Neurodevelopmental Disorders/genetics* ; Kinesins/genetics*

Objective:To characterize the histopathological subtypes and their clinicopathological parameters of gender and onset age by common, rare and sparse primary esophageal malignant tumors (PEMT).Methods:A total of 272 437 patients with PEMT were enrolled in this study, and all of the patients were received radical surgery. The clinicopathological information of the patients was obtained from the database established by the State Key Laboratory of Esophageal Cancer Prevention & Treatment from September 1973 to December 2020, which included the clinical treatment, pathological diagnosis and follow-up information of esophagus and gastric cardia cancers. All patients were diagnosed and classified by the criteria of esophageal tumor histopathological diagnosis and classification (2019) of the World Health Organization (WHO). The esophageal tumors, which were not included in the WHO classification, were analyzed separately according to the postoperative pathological diagnosis. The χ 2 test was performed by the SPSS 25.0 software on count data, and the test standard α=0.05. Results:A total of 32 histopathological types were identified in the enrolled PEMT patients, of which 10 subtypes were not included in the WHO classification. According to the frequency, PEMT were divided into common (esophageal squamous cell carcinoma, ESCC, accounting for 97.1%), rare (esophageal adenocarcinoma, EAC, accounting for 2.3%) and sparse (mainly esophageal small cell carcinoma, malignant melanoma, etc., accounting for 0.6%). All the common, rare, and sparse types occurred predominantly in male patients, and the gender difference of rare type was most significant (EAC, male∶ female, 2.67∶1), followed with common type (ESCC, male∶ female, 1.78∶1) and sparse type (male∶ female, 1.71∶1). The common type (ESCC) mainly occurred in the middle thoracic segment (65.2%), while the rare type (EAC) mainly occurred in the lower thoracic segment (56.8%). Among the sparse type, malignant melanoma and malignant fibrous histiocytoma were both predominantly located in the lower thoracic segment (51.7%, 66.7%), and the others were mainly in the middle thoracic segment.Conclusion:ESCC is the most common type among the 32 histopathological types of PEMT, followed by EAC as the rare type, and esophageal small cell carcinoma and malignant melanoma as the major sparse type, and all of which are mainly occur in male patients. The common type of ESCC mainly occur in the middle thoracic segment, while the rare type of EAC mainly in the lower thoracic segment. The mainly sparse type of malignant melanoma and malignant fibrous histiocytoma predominately occur in the lower thoracic segment, and the remaining sparse types mainly occur in the middle thoracic segment.

Objective To investigate the association of energy metabolic markers with the risk of spontaneous bacterial peritonitis (SBP) in patients with decompensated hepatitis B virus-related liver cirrhosis (HBV-LC). Methods A retrospective analysis was performed for the clinical data of the patients with decompensated HBV-LC who were admitted to Mengchao Hepatobiliary Hospital of Fujian Medical University from November 2017 to November 2019, and baseline clinical parameters and energy metabolic markers were compared between the patients with SBP and those without SBP within 2 weeks after admission. A multivariate logistic regression analysis was performed to investigate the risk factors for SBP. The t -test was used for comparison of normally distributed continuous data between two groups, and the Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between two groups; the Fisher's exact test was used for comparison of categorical data between two groups. The receiver operating characteristic (ROC) curve was plotted to evaluate the diagnostic efficiency of the newly established logistic regression model, and with the corresponding point of Youden index as the cut-off value, the DeLong test was used to compare the area under the ROC curve (AUC). Results A total of 50 patients with decompensated HBV-LC were included, among whom 23 (46%) developed SBP within 2 weeks after admission and 27 (54%) had no SBP during hospitalization. Compared with the non-SBP patients, the SBP patients had significantly lower triglyceride, prealbumin, and prothrombin time activity (PTA) and significantly higher international normalization ratio, C-reactive protein (CRP), and Model for End-Stage Liver Disease score (all P < 0.05). Comparison of baseline energy metabolic markers showed that compared with the non-SBP patients, the SBP patients had significantly lower respiratory quotient (RQ) [0.79(0.76-0.86) vs 0.85(0.79-0.91), P =0.041] and carbohydrate oxidation (CHO) rate [20.50%(15.25%-41.05%) vs 41.6%(22.25%-68.05%), P =0.041]. The multivariate logistic regression analysis showed that PTA was an independent risk factor for SBP in the patients with decompensated HBV-LC during hospitalization (odd ratio=0.004, P =0.008), and the regression model established based on the variables including PTA, CRP, RQ, and CHO had an AUC of 85.0% and a cut-off value of 0.60 at the maximum Youden index, with a specificity of 85.19% and a sensitivity of 73.91%, suggesting that this model had a better discriminatory ability than CRP (AUC=74.5%, P =0.049) and procalcitonin (AUC=56.4%, P < 0.01). Conclusion There are significant reductions in the energy metabolic markers RQ and CHO in the patients with decompensated HBV-LC who develop SBP within a short term, and their combination with PTA, CRP, and CHO/RQ ratio can help clinicians identify the patients at a high risk of SBP in the early stage and enhance nutrition support for such patients.

Objective:To investigate and analyze the energy metabolism characteristics and the correlation between energy metabolism and the risk of secondary bacterial infection in patients with hepatitis B virus-related chronic liver disease (HBV-CLD).Methods:Data of 183 cases admitted to the Mengchao Hepatobiliary Hospital of Fujian Medical University from November 2017 to November 2020 were retrospectively analyzed. 79 cases of chronic hepatitis B, 51 cases of hepatitis B-related liver cirrhosis, and 53 cases of hepatitis B-related liver failure were collected. Among them patients with liver failure and decompensated liver cirrhosis were defined as severe liver disease group. The Quark RMR indirect calorimetry (COSMED Corporation, Italy) was used to exam the patients' energy metabolism condition, and the incidences of secondary bacterial infection of the patients during hospitalization were recorded. Shapiro-Wilk test and normal QQ plot were used to analyze the normal distribution of continuous variable data, which was consistent with the normal distribution and was described by mean ± standard deviation. In addition, if it did not conform to the normal distribution, the median and interquartile distance were used to describe it. Levene’s test was used to test the homogeneity of variance of the data, which was consistent with the normal distribution. The t-test was used to compare the means of the two groups of samples. One-way analysis of variance was used to compare the mean values of the three groups of samples, and then the Tukey's test was used to compare the two groups. If the variance was uneven or did not conform to the normal distribution, the Wilcoxon rank sum test was used to compare the differences between the two groups. Kruskal-Wallis test (H test) was used to compare the differences between the three groups of samples, and then the Dunnett’s test (Z test) was used for comparison between the two groups. Categorical variable data were analyzed using chi-square test. Logistic regression analysis was used to screen independent risk factors, and the criteria for variable inclusion ( P < 0.05). Results:The respiratory entropy (RQ) and non-protein respiratory entropy (npRQ) of the three groups had statistically significant difference ( P < 0.05). Among them, the RQ and npRQ of the chronic hepatitis B group were higher than hepatitis B-related liver cirrhosis group and hepatitis B-related liver failure group. There were statistically significant differences in fat oxidation rate (FAT%) and carbohydrate oxidation rate (CHO%) between the three groups ( P < 0.05). Compared with hepatitis B-related liver cirrhosis group and hepatitis B-related liver failure group, chronic hepatitis B group ( P < 0.05) had lower FAT% and higher CHO%. There were no statistically significant differences in the measured and predicted resting energy expenditure and protein oxidation rate (PRO%) between the three groups. The incidence of secondary bacterial infection in patients with severe liver disease was 48.39% (45/93). Compared with the non-infected group, the RQ and npRQ values ??of the infected group were significantly decreased ( P < 0.05), while FAT% was significantly increased ( P < 0.05). Logistic regression analysis showed that glutamyltransferase, cholesterol, and npRQ were independent risk factors for secondary bacterial infections in patients with severe liver disease. Glutamyltransferase elevation, and cholesterol and npRQ depletion had suggested an increased risk of secondary bacterial infection. Subgroup analysis of patients with hepatitis B-related liver failure also showed that compared with non-infected group, RQ value and npRQ value of secondary bacterial infection group were significantly decreased ( P < 0.05), while FAT% was significantly increased ( P < 0.05). Conclusion:Patients with hepatitis B virus-related chronic liver disease generally have abnormal energy metabolism. Low RQ, npRQ, CHO% and high FAT% are related to the severity of the disease; while npRQ reduction is related to the risk of secondary bacterial infection in patients with severe liver disease, and thus can be used as a clinical prognostic indicator.

OBJECTIVE： To establish HPLC fingerprints of Paeonia tactilora decoction pieces， and to conduct its cluster analysis and principal component analysis. METHODS： HPLC method was adopted. The determination was performed on SunFire® C18 column with mobile phase consisted of acetonitril-0.05％ phosphoric acid solution （gradient elution） at the flow rate of 1.0 mL/min. The detection wavelength was set at 230 nm， the column temperature was 30 ℃， the collection time was 70 min，and sample size was 15 μL. Using paeoniflorin as reference， HPLC fingerprints of 26 batches P. tactilora decoction pieces from different habitats and 30 batches by different processed methods were established. The similarity of samples was evaluated by TCM Chromatographic Fingerprint Similarity Evaluation System （2012 edition） to confirm common peak. Cluster analysis and principal component analysis were performed by using SPSS 20.0 software. RESULTS： There were 9 common peaks in HPLC fingerprints of 26 batches of sample from different habitats， the similarity of which was higher than 0.880. Six peaks were identified， including gallic acid， catechin， albiflorin， paeoniflorin， 1，2，3，4，6-pentagalloylglucose and benzoylpaeoniflorin. Cluster analysis showed that 26 batches of samples were clustered into 2 categories when cosine distance was 15. S1-S21 were clustered into one category； S22-S26 were clustered into the other category. By principal component analysis， the accumulative contribution rate of two main components was 81.124％. There were 10 common peaks in HPLC fingerprints of 30 batches of sample by different processed methods， the simi- larity of which was higher than 0.970. Seven peaks were identified， including gallic acid， catechin， aplopaeonoside， albiflorin， paeoniflorin， 1，2，3，4，6-pentagalloylglucose and benzoylpaeoniflorin. Cluster analysis showed that 30 batches of samples were clustered into 2 categories when cosine distance was 25. B1-B10 were clustered into one category； C1-C10 and J1-J10 were clustered into the other category. By principal component analysis， the accumulative contribution rate of four main components was 86.887％. CONCLUSIONS： Established HPLC fingerprint， the results of cluster analysis and principal component analysis can provide reference for quality control of decoction pieces of P. tactilora.

Objective To investigate the effects of alcohol extract of Kunmu decoction on proliferation and apoptosis of rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS). Methods Synovial tissues were obtained from patients with active RA received joint replacement or arthroscopy. The surface antigen and the amount of apoptotic cells were determined by flow cytometry. The inhibitive effect was detected by MTT assay. Results The CD90+surface antigen of synoviocytes was (94.78 ± 0.98)%. The inhibitive effect on the proliferation in all treatment groups were in a time-and dose-dependent manner. The apoptosis rate was increased in a dose-dependent manner among all dosage alcohol extract groups. Conclusion Kunmu decoction might inhibit proliferation and induce apoptosis of RA-FLS.

Objective:To study the relationship between serum soluble intercellular adhesion molecule-1(sICAM-1) and vascular endothelial growth factor(VEGF), placenta growth factor(PLGF), vascular endothelial growth factor receptor 1(VEGFR1, Flt-1), soluble vascular endothelial growth factor receptor 1(sVEGFR1, sFlt-1) mRNA expression in placenta tissue of preeclampsia(PE). Methods: The serum level of sICAM-1 was measured by enzyme-linked immunosorbent assay and gene expression of placenta tissue was detected by quantitative reverse transcriptase-polymerase chain reaction(RT-PCR). Results:(1)The average serum level of sICAM-1 was(218.45±29.93) μg/L in PE group compared with (168.84±19.39) μg/L in controls(P < 0.01).(2)The mRNA expressions of VEGF, PLGF, Flt-1 and sFlt-1 were increased in placenta of PE than those in controls (P < 0.01).(3)There was a positive correlation between the serum level of sICAM-1 and the sFlt-1 mRNA expression in placenta tissue(r = 0.90, P < 0.01). Conclusion: The serum level of sICAM was increased in the patients with PE. The expressions of VEGF, PLGF, Flt-1 and sFlt-1mRNA were increased remarkably in the placenta tissue of palients with PE,especial for sFlt-1. The remarkable increase for expression of sFlt-1mRNA may contribute to endothelial dysfunction, hypertension and proteinuria.

Myelin of the adult mammalian central nervous system(CNS)has been attributed to affect nerve structural plasticity and suppress regeneration of nerve fibers.Nogo-A is possibly the best characterized of a variety of neurite growth inhibitors in CNS myelin.Neutralizing its activity results in improved axon regrowth and functional recovery in experimental spinal cord injury(SCI)models of animals.Nogo-A and its receptors,especially Nogo-66 receptor(NgR),p75 neurotrophin receptor(p75NTR),and LINGO-1 increasingly become the hot spot in the study of SCI repair,and have become the major targets for therapeutic intervention to promote axon regeneration after SCI.Inhibition of Nogo-A and its receptors NgR/p75NTR/LINGO-1 may be promote the regeneration of axon and maximize functional recovery after SCI.

Objective To establish a rapid test method for the determination of five sweeteners and two preservatives in the milk beverage by HPLC.Methods Trough a set of pretreatment such as dilution,protein precipitation,high-speed centrifugation,the samples were tested.With NUCLEODUR C18 RP-column separation,phosphate buffer(pH=5.0) and acetonitrile were used as the mobile phase,gradient elution analysis was conducted,the detective wavelength was 205nm.Results The separation of one sample was achieved within 15 min,five sweeteners and two preservatives could be well separated,the linear range was 5.00-100.0 mg/L,r≥0.9993,RSD was 1.5%-4.2%(n=6),the average recovery rate was 85.2%-108%.Conclusion This method is simple and quick with better reproducibility and selectivity,it is an effective way to determine sweeteners and preservatives in milk beverage.