Objective This study was to investigate the relation of nitric oxide synthase with cell proliferation and apoptosis in hepatocellular carcinoma (HCC). Methods The expression and localization of inducible nitric oxide synthase (iNOS) and PCNA in 21 patients with HCC were studied by immunohistochemistry. Apoptotic cell in tissue section was detected by TUNEL (terminal deoxynucleotidyl transferase [TdT] mediated desoxyuridinetriphosphate [dUTP] nick end labeling). Results The iNOS protein was markedly expressed in the liver tissue of hepatic cirrhosis or chronic liver hepatitis, which was close to tumor edge. Meanwhile it was weakly or undetectable in the liver tissue localized beyond 1.5 cm from the tumor edge. The iNOS protein was markedly expressed at the invasive tumor edge and HCC cells that invased into stroma, but it was weakly or undetectable at the most tumor core. The expression of iNOS was correlated with the proliferative activity of cells, and the number of apoptotic cells was correlated to the number of PCNA positive cells ( P

Objective To analyze the distribution of B-cell subtypes and IgG subclasses in pa-tients with non-small cell lung cancer ( NSCLC) and to investigate their potential functions in the progress of NSCLC.Methods Flow cytometry analysis was used to detect the distribution of immature B cells, memory B cells and mature B cells in 25 healthy subjects and 55 patients who were at different clinical stages of NSCLC.Enzyme-linked immune-sorbent assay ( ELASA) was performed to measure the concentrations of IgG1, IgG2, IgG3, IgG4 and total IgG, as well as the ratio of each IgG subclass in total IgG.Results The percentages of immature and memory B cells in CD19+B cells were positively correlated with the progress of NSCLC.The ratio of IgG4/IgG gradually increased along with the development of NSCLC, while that of the other three IgG subclasses showed no significant changes as compared with that of the control group.Conclu-sion The imbalanced distribution of B-cell subtypes and the up-regulated ratio of IgG4/IgG were closely as-sociated with the development of NSCLC.This study paved the way for further investigation on more effective immune therapy strategies targeting NSCLC.

0.05), but not in other brain regions. The number of GFAP-immunopositive cells of the E.coli-treated pups was markedly increased in periventricular white matter and hippocampus at P7 compared with the control group (P0.05). CONCLUSION: Intrauterine infection induces an increased expression of GFAP in the neonatal brain. [

In this experiment, a novel biotin-avidin conjugation probe was synthesized and employed in the detection of reverse-phase protein microarray. Firstly, the proportion of the biotin-avidin conjugation probe was optimized. Then the rat IgG and goat anti-rat IgG system was served as a model to optimize the fabrication conditions of reverse-phase protein microarray, including the non-specific absorption of streptavidin-Cy3 molecules, spotting buffer as well as protein activities. At last, the biotin-avidin conjugation probe was applied to the detection of the reverse-phase protein microarray. The results show that the protein microarray prepared by using BSA spotting buffer could prevent non-specific absorptions of fluorescent molecules and improve the sensitivity, effectively. In addition, compared with traditional biotin-avidin system, the detection limit could be improved four times using the biotin-avidin conjugation probe. In conclusion, the biotin-avidin conjugation probe has its merits of easy synthesis, low price and could be further conjugated with other signal amplification techniques, which is promising to be used in the detection of protein microarray.
Avidin ; chemistry ; Biotin ; chemistry ; DNA Probes ; Immunoglobulin G ; analysis ; immunology ; Protein Array Analysis ; methods

Objective:To summarize the clinical features and PLGL gene variation characteristics of children with CHIME syndrome. Methods:The medical records of one patient who was diagnosed with CHIME syndrome in the Third Affiliated Hospital of Zhengzhou University in October 2018 were analyzed.Foreign and domestic databases were searched with " CHIME syndrome or PIGL gene" as the keywords, so as to review clinical features of CHIME syndrome and PIGL gene variation characteristics. Results:(1) The boy, 1 year old and 3 months, developed seizures at the age of 7 months, when he received rehabilitation due to developmental delay.Physical examination showed that the boy had facial dysmorphisms, including high forehead, ocular hypertelorism, low and flat nasal root, broad nose tip, full lips, overfolded helices, cleft palate, developmental delay, dry skin, erythematous papular rash on the neck, and indirect inguinal hernia. Conductive deafness was revealed by the hearing test and retinal defect was found in fundus examination.Whole exome sequencing test identified PIGL(NM_004278)gene compound hybrid variation.The frameshift variation c. 26delT was present in one allele, combined with a synonymous variation c. 333C>T in the opposite allele.(2) A total of 9 CHIME syndrome patients were retrieved from the databases.No cases were reported in China.All 9 patients had craniofacial dysmorphism, epilepsy, conductive deafness, development delay and retinal defect.Eight patients had ichthyosiform skin, 6 patients had congenital heart disease and 4 patients had renal malformation.There were 6 different kinds of PIGL gene variations in patients, including 7 missense variants, 4 frameshift variants, 3 deletion variants, 2 nonsense variants, 1 splice variant, and 1 synonymous variant. All of the missense variants were c. 500T>C (p.Leu167Pro), which was the most common site. Conclusions:CHIME syndrome is mainly manifested by nervous system and dermal system abnormalities, and often involves multiple systems. PIGL gene variation is the cause of CHIME syndrome, and c. 500T>C (p.Leu167Pro) is the most common site.

In order to ensure the biosafety of the IFN-gamma antiviral activity assay, we used a replication-deficient VSV carrying GFP as an interferon sensitive indicator virus (VSVdeltaG*G). The antiviral activities of porcine IFN-gamma expressed in Escherichia coli and in baculovirus on MDBK cells were assessed. The results showed that the antiviral activity of porcine IFN-gamma expressed in baculovirus could reach 10(5) IU/mL, while the porcine IFN-gamma expressed in E. coli showed some antiviral activity (32 IU/mL) after refolding. The results of the VSVdeltaG*G-based antiviral assay were almost identical to that of the VSV*GFP-based assay, suggesting it is highly feasible to use VSVdeltaG*G as a substitute for VSV*GFP, making assays for IFN-gamma antiviral activity safer and more accurate.
Animals ; Antiviral Agents ; pharmacology ; Baculoviridae ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Interferon-gamma ; biosynthesis ; genetics ; metabolism ; pharmacology ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Swine ; Vesiculovirus ; drug effects ; physiology

Objective:To investigate the current status of job burnout among trainees in the standardized residency training of pediatrics (short for "pediatric residents") in the hope of identifying the problems involved, and proposing corresponding countermeasures and suggestions based on these current situations and problems.Methods:This study conducted a questionnaire survey among the pediatric residents who received the standardized residency training in The Children's Hospital, Zhejiang University School of Medicine. The questionnaire consisted of two parts: general information survey and Maslach job burnout survey. The generated data were analyzed using IBM SPSS 26.0. Chi-square test was used to determine the equilibrium of the groupings for the basic situation of the data. If the normal distribution of the data in two groups was satisfactory, the t-test was used for inter-group comparison; otherwise, the Kruskal-Wallis H test was employed. For more than two groups, if the data met the normal distribution and the variance was homogeneous, one-way analysis of variance was used for inter-group comparison; otherwise, the Kruskal-Wallis H test was used. Multiple linear regression analysis was used for multivariate analysis. Results:A total of 138 pediatric residents participated in this survey. The detection rate of job burnout was 65.2% (90/138), of which 45 (32.6%), 29 (21.0%) and 16 (11.6%) were mild, moderate and severe, respectively. In univariate analysis, there was a statistically significant difference in job burnout among pediatric residents in different grades ( H=7.22, P=0.027), and the low achievement score (27.90±8.48) of the first-year pediatric residents was higher than that of the second-year pediatric residents (23.54±6.79), and the difference was statistically significant ( t=-2.25, P=0.025). Multiple linear regression analysis regression model showed no statistically significant difference. Conclusion:The level of job burnout among pediatric residents is high, and relevant departments should pay attention to the problem of job burnout of the pediatric residents, and take various measures to prevent the occurrence of job burnout.

Tripterygium glycosides tablet(TGT),the classical commercial drug of Tripterygium wilfordii Hook.F.has been effectively used in the treatment of rheumatoid arthritis,nephrotic syndrome,leprosy,Behcet's syndrome,leprosy reaction and autoimmune hepatitis.However,due to its narrow and limited treatment window,TGT-induced organ toxicity(among which liver injury accounts for about 40％of clinical reports)has gained increasing attention.The present study aimed to clarify the cellular and molecular events underlying TGT-induced acute liver injury using single-cell RNA sequencing(scRNA-seq)technology.The TGT-induced acute liver injury mouse model was constructed through short-term TGT exposure and further verified by hematoxylin-eosin staining and liver function-related serum indicators,including alanine aminotransferase,aspartate aminotransferase,alkaline phosphatase and total bilirubin.Using the mouse model,we identified 15 specific subtypes of cells in the liver tissue,including endothelial cells,hepatocytes,cholangiocytes,and hepatic stellate cells.Further analysis indicated that TGT caused a significant inflammatory response in liver endothelial cells at different spatial locations;led to marked inflammatory response,apoptosis and fatty acid metabolism dysfunction in hepatocytes;activated he-patic stellate cells;brought about the activation,inflammation,and phagocytosis of liver capsular macrophages cells;resulted in immune dysfunction of liver lymphocytes;disturbed the intercellular crosstalk in liver microenvironment by regulating various signaling pathways.Thus,these findings elaborate the mechanism underlying TGT-induced acute liver injury,provide new insights into the safe and rational applications in the clinic,and complement the identification of new biomarkers and ther-apeutic targets for liver protection.

This paper reported a case of neonatal radial nerve palsy that was successfully treated by conservative therapy. The patient with 39 weeks of gestational age was born vaginally. On the 2nd day after birth, right wrist drop, decreased muscle strength in the right upper limb, the erythema patch, and the subcutaneous nodule on the upper arm were observed. Electromyography revealed acute denervation of the radial nerve. Based on the electromyography results combined with clinical evaluations, neonatal radial nerve palsy was diagnosed. The patient was treated with self-made simple splint fixation along with comprehensive treatment. At 15 days of age, the family members removed the splint fixation themselves (13 days of fixation). At the age of 20 days, the symptoms of right wrist drop had disappeared, and the grasp reflex and Moro reflex of both hands returned to normal. A follow-up electromyography conducted at six months after discharge showed no obvious abnormalities.

To explore the effects of intrauterine infection on early growth and neurobehavioral development in neonatal rats. (E. coli) was inoculated into uterine cervix of pregnant rats with gestation of 15 d to establish the intrauterine infection model, and the effect on the delivery of pregnant rats was observed. The neonatal rat brain tissue was stained with Hematoxylin-Eosin and the cerebral white matter damage was assessed. Immunohistochemical staining and Western blot analysis were performed to evaluate the expression of glial fibrillary acidic protein (GFAP), 2', 3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) and neurofilament (NF) in pup brains. Birth weight and early growth development indices were monitored,and neurobehavioral tests were performed to access the change of neurobehavioral development in neonatal rats. The white blood cell count increased significantly in the uterus and placenta of the pregnant rats after intrauterine E. coli infection and no significant impact was observed on the delivery of pregnant rats. Weak staining and focal rarefaction of cerebral white matter from rats at P7 in intrauterine infection group were observed. The expression of GFAP markedly increased (<0.05) in infection group, while the level of CNPase and NF in pup brains at P7 significantly decreased (<0.05 or <0.01). Compared with control group, the neonatal rats in infection group had lower birth weight and slower weight gain during the suckling period (<0.05 or <0.01), and the completion times of ear opening, eye opening, surface righting, negative geotaxis, acoustic startle and swimming test in infection group were significantly delayed (<0.05 or <0.01). Intrauterine infection in pregnant rats can induce cerebral white matter damage and retardation of early growth and neurobehavioral development in neonatal rats.
Animals ; Animals, Newborn ; Behavior, Animal ; Body Weight ; Disease Models, Animal ; Escherichia coli ; Escherichia coli Infections ; complications ; physiopathology ; Female ; Glial Fibrillary Acidic Protein ; genetics ; Growth Disorders ; etiology ; Leukoencephalopathies ; etiology ; Pregnancy ; Pregnancy Complications, Infectious ; physiopathology ; Rats ; Rats, Sprague-Dawley