Objective: To investigate the efficacy, safety, and traditional Chinese medicine (TCM) medication patterns of rituximab (RTX) combined with TCM on treating children with steroid-dependent nephrotic syndrome (SDNS). Methods: One hundred and forty-three children with SDNS who visited the Pediatric Nephrology Department of the First Affiliated Hospital of Henan University of Chinese Medicine from January 2018 to December 2022 were enrolled. A cohort study design was adopted, with " RTX treatment" as the exposure factor. Children who met this exposure factor were assigned to the RTX cohort (RTX, glucocorticoid, immunosuppressive agent, combined with traditional Chinese medicine syndrome differentiation treatment), whereas those who did not were assigned to the basic treatment cohort (glucocorticoid, immunosuppressive agent, combined with traditional Chinese medicine syndrome differentiation treatment ), and followed up for 6 months. The frequency of urinary protein recurrences, urinary protein remission duration, proportion and duration of steroid reduction and cessation, cumulative usage of steroids, proportion of recurrence, recurrence amount of steroid used, efficacy of TCM syndrome, and laboratory and safety indicators after treatment, and height and CD19+ B cell count before and after treatment were compared between the two cohorts. The medication patterns of TCM in the two cohorts were analyzed using frequency statistics, association rule analysis, and systematic clustering analysis. Results: Compared with the basic treatment cohort, the RTX cohort showed a decrease in the frequency of urinary protein recurrence, extended sustained remission of urinary protein, an increase in the proportion of steroid reduction and cessation, a shorter duration of steroid reduction and cessation, a decrease in cumulative steroid dosage, a lower recurrence rate, a decrease in CD19+ B cell count, and a decrease in 24-h urinary total protein quantification and the level of cholesterol (P<0.05). No significant difference in the recurrence amount of steroid used, height, TCM syndrome efficacy, albumin, aspartate transaminase, blood urea nitrogen, platelet count, and safety indicators between the two cohorts. Children with SDNS were mostly characterized by qi and yin deficiency syndrome, followed by spleen and kidney yang deficiency syndrome. A total of 175 TCMs were included, including 28 high-frequency drugs such as Huangqi, Fuling, Gancao, Baizhu, Dangshen, and Jiuyurou. The primary use of medication is to nourish the qi and spleen, nourish the kidney, and warm yang. The analysis of association rules yielded eight binary associations and ten three-phase associations, with Huangqi, Baizhu, Fuling, and Dangshen, being the most closely related. Cluster analysis identified four TCM combinations, primarily focusing on tonifying kidney and replenishing essence, benefiting qi and nourishing yin, and removing blood stasis. Conclusion RTX combined with TCM syndrome differentiation treatment can reduce the recurrence frequency of SDNS, prolong the remission period, reduce the glucocorticoid dosage, and have no marked effect on height growth. No apparent adverse reactions were observed. TCM should focus on nourishing qi and yin while removing blood stasis.

Following the principles of evidence-based medicine,in accordance with the structure and drafting rules of standardized documents,based on literature research,according to the characteristics of chronic cough in children and issues that need to form a consensus,the TCM Guidelines for Diagnosis and Treatment of Chronic Cough in Children was formulated based on the Delphi method,expert discussion meetings,and public solicitation of opinions.The guideline includes scope of application,terms and definitions,eti-ology and diagnosis,auxiliary examination,treatment,prevention and care.The aim is to clarify the optimal treatment plan of Chinese medicine in the diagnosis and treatment of this disease,and to provide guidance for improving the clinical diagnosis and treatment of chronic cough in children with Chinese medicine.

Objective To investigate the syndrome differentiation characteristics of children with atopic dermatitis(AD)and the immune imbalance status in children with different syndrome types of AD.Methods A total of 159 AD children and 100 normal control children were enrolled.The peripheral blood eosinophil(Eo)count was measured by impedance method,total serum immunoglobulin E(IgE)by immunoturbidimetric assay,and interferon-gamma(IFN-γ),interleukin-4(IL-4),interleukin-5(IL-5)and interleukin-17(IL-17)were measured by multiple microspheres flow immunofluorescence assay.Results Among 159 AD children,syndrome of heart-fire and spleen-deficiency was most commom,accounting for 38.4%,followed by syndrome of blood-deficiency and wind-dryness(22.0%),syndrome of heat accumulation in heart and spleen(20.1%)and syndrome of spleen-deficiency and dampness-accumulation(19.5%).Compared with normal control group,there was no significant difference in serum IFN-γ level among different syndrome types of AD.The levels of peripheral blood Eo,serum total IgE,IL-4 and IL-17 in AD with heart-fire and spleen-deficiency syndrome were significantly increased(P<0.05).The levels of peripheral blood Eo,IL-4,IL-5 and IL-17 in AD with blood-deficiency and wind-dryness syndrome were significantly increased(P<0.05).The levels of IL-4,IL-5 and IL-17 in AD with heat accumulation in heart and spleen syndrome were significantly increased(P<0.05).The levels of peripheral blood Eo and serum IL-4 in AD with spleen-deficiency and dampness-accumulation syndrome were significantly increased(P<0.05).Conclusion Heart-fire and spleen-deficiency syndrome is the most common type in children with AD,however,the main type under 3 years old is heat accumulation in heart and spleen syndrome.Th2/Th17 immune imbalance are the main pathogenesis in heart-fire and spleen-deficiency syndrome,blood-deficiency and wind-dryness syndrome and heat accumulation in heart and spleen syndrome,and Th2 immune imbalance is the main pathogenesis of spleen-deficiency and dampness-accumulation syndrome.

Objective:To analyze the expression of cell division cycle associated protein 5 (CDCA5) in pancreatic cancer tissues and its correlation with prognosis based on the bioinformatics.Methods:The RNA sequencing data (HTSeq-FPKM) and corresponding clinical information of 168 pancreatic cancer samples from January to December 2021 were downloaded from the Cancer Genome Atlas (TCGA) database, and the data of 179 pancreatic patients from January to December 2021 were downloaded from the GEPIA2 database, and 171 normal pancreatic tissues from TCGA and GTEx databases were simultaneously integrated. The relative expression level of CDCA5 mRNA in pancreatic cancer patients in GEPIA2 database and its relationship with overall survival (OS) and disease-free survival (DFS) were explored. Combined with the clinical data of the patients, univariate and multivariate Cox regression model analysis was used to analyze the factors influencing the OS of pancreatic cancer patients. Gene set enrichment analysis (GSEA) was performed to investigate the possibly involved signal pathways of CDCA5 in pancreatic cancer.Results:In the GEPIA2 database, the relative expression level of CDCA5 mRNA in pancreatic cancer tissues was higher than that in normal pancreatic tissues, and the difference was statistically significant ( P < 0.05). The pancreatic cancer patients were divided into the high CDCA5 mRNA expression group (89 cases) and the low CDCA5 mRNA expression group (89 cases) according to the median of relative expression level of CDCA5 mRNA (the case equal to the median value was not subgrouped). Survival analysis showed that patients with high CDCA5 mRNA expression had shorter OS ( P = 0.024) and DFS ( P = 0.025) compared with those with low CDCA5 mRNA expression. Multivariate Cox analysis showed that in TCGA database, N staging ( HR = 2.15, 95% CI 1.24-3.72, P = 0.006) and CDCA5 expression ( HR = 1.71, 95% CI 1.23-2.38, P = 0.001) were independent influencing factors of OS for pancreatic cancer patients. The results of GSEA enrichment analysis indicated that high CDCA5 mRNA expression was enriched in 13 biological pathways [all P < 0.05, false discovery rate (FDR) < 0.005] including cell cycle, DNA replication, homologous recombination, pyrimidine metabolism, mismatch repair, pentose phosphate pathway, glycolysis gluconeogenesis and p53. The expression of CDCA5 mRNA was positively correlated with the expressions of HK2, PKM, PGK1, ALDOA, EN01 and LDHA (all P < 0.05). Conclusions:CDCA5 is highly expressed in pancreatic cancer tissues and is associated with poor prognosis of patients, and it can be used as a prognostic marker for pancreatic cancer.

Objective: To establish a method for the determination of manganese in urine by graphite furnace atomic absorption spectrometry (AAS) without the use of matrix modifier. Methods: The urine samples were 5 times diluted with 1% nitric acid then directly determined by AAS. Zeeman was used for background correction. Results: The linear range for determination of manganese in urine was 5~60 μg/L (urine) . The correlation coefficient was greater than 0.995 with the detection limit of 1.5 μg/L and with the lower limit of quantification of 5.0 μg/L. The relative standard deviations (RSDs) of within-run precision was between 1.1%~4.3%, the RSDs of between-run precision was between 3.3%~7.0%. The average recovery was 102.6%. The samples can be stored for 14 days at room temperature, 4℃, -8 ℃ and -35 ℃. Conclusion The method is feasible for determination of manganese in urine.

Objective To establish a method for the isolation and identification of platelets.Methods 10 healthy volunteers were selected to collect the EDTA anticoagulant venous blood of 3 tubes,each tube was 2 ml,which was divided into the whole blood cell tube,platelet rich plasma (control group),and stepped centrifugal platelet extract (experiment group).Platelet was isolated by simple centrifugation method(PRP) and stepped centrifugal method.The two groups were full blood count and analyzed by microscopic morphology and platelet activity test.Leukocyte specific HGB gene and platelet mitochondrial ND1 gene content was analyzed by real time PCR.Results Platelets were extracted and detected in control group and experimental group.Platelets were found and white blood cells and red blood cells were not remained in experimental group.Platelets and sporadic white blood cells were found in control group.The platelet pick up rate of experiment group was significantly higher than control group,the difference was statistically significant.Experimental gene content HGB of experiment group was significantly lower than control group,the difference was statistically significant (t=-3.281,-2.865,P＜0.05).ND1 gene content of experiment group higher than the control group,the difference was not statistically significant.There was no significant difference for platelet activity test between experimental group and control group (t=-0.046,-0.799,P＞ 0.05).Conclusion A isolation and identification method of stepped centrifugal platelet was established.The method can be used for the study of platelet gene and the functional analysis of platelets.

Objective: To investigate the detection of a human leukocyte antigen-B (HLA-B) allele HLA-B*13:01 by dual allele-specific real-time polymerase chain reaction (PCR) in patients with trichlorethylene-induced dermatitis. Methods: A total of 20 patients with trichlorethylene-induced dermatitis who were admitted and treated from January 2014 to October 2016 were enrolled as case group, and 20 persons who underwent physical examination from January to October, 2016 were enrolled as control group. Peripheral cubital venous blood samples were collected from all subjects, and dual allele-specific real-time PCR was used to detect the HLA-B*13:01 gene. The two groups were compared in terms of the proportion of subjects carrying HLA-B*13:01 gene. Results: There were no significant differences between the case group and the control group in median age (25.0 years vs 27.0 years, Z=0.30, P>0.05) and the proportion of male subjects (60.0% vs 70.0%, χ²=0.44, P>0.05) . The mean time of exposure to trichloroethylene was 30.8 days in the case group, while the subjects in the control group were not exposed to trichloroethylene. The case group had a significantly higher frequency of HLA-B*13:01 gene than the control group (80.0% vs 20.0%, χ²=14.40, P<0.01) with an odds ratio of 16.00. Conclusion Dual allele-specific real-time PCR can be used for detection of the HLA-B*13:01 gene in patients with trichlorethylene-induced dermatitis.

Objective Total aortic arch replacement and stented elephant trunk surgery is an important surgical method for acute type A aortic dissection, and the short, middle, long term curative effect has been recognized by more and more experts at home and abroad.Circulatory arrest was an independent risk factor for postoperative complications and mortality in patients with type A aortic dissection.The aim of this article is to observed the effection of a new technology to block aortic arch, whicn can shorten the circulatory arrest time to 2 minutes and avoid harm of circulatory arrest on patients.Methods From May 2016 to February 2017, 68 patients with acute type A aortic dissection were divided into the conventional group and the modified group.All the patients underwent total arch replacement and stented elephant trunk surgery.The rectal temperature of the conventional group was 25℃ and circulatory arrest time was 20 min.While the rectal temperature of the modified group was 28℃ and and circulatory arrest time was 2 min.Results In the conventional operation group, 3 patients died after operation while all the patients in the modified group were cured and discharged.There are no differences between the two groups in the time of cardiopulmonary bypass(CPB) and heart arrest time(P>0.05).There are Significant differences in CPB time, circulatory arrest time, postoperative awake time, intubation time, amount of blood used, the amount of drainage during the first two days after operation, the time staying in ICU and the postoperative time in hospital.And the modified group was much better.(P<0.05)Conclusion The results of new technology blocking aortic arch in the patients with acute type A aortic dissection are better than the conventional surgical approach during the perioperative period.This technology is simple and effective.It is worth promoting.

OBJECTIVE: To explore the correlation between human leukocyte antigen( HLA)-B~* 13 : 01 allele and liver dysfunction in patients with occupational medicamentosa-like dermatitis due to trichloroethylene( OMDT). METHODS: Twenty patients with OMDT were chosen as study subjects by using a convenient sampling method. The sequence-based genotyping method was used for detecting HLA-B~* 13 : 01 allele in the DNA samples from peripheral blood of all study subjects. The serum levels of total protein,albumin,total bilirubin,direct bilirubin and alanine aminotransferase,aspartate aminotransferase and alkaline phosphatase activities in patients were examined. The correlation between the number of HLA-B~* 13 : 01 alleles and the liver function indices was also analyzed. RESULTS: There were 16 patients carrying HLA-B~* 13: 01 allele. The serum total protein in the HLA-B~* 13: 01 carriers was higher than that of non-carriers( P < 0. 05). The serum total protein was positively correlated with the number of patients carrying HLA-B~* 13: 01 alleles( P < 0. 05). CONCLUSION: The degree of liver function damage in OMDT patients may be related to carrying the HLA-B~* 13: 01 allele.

OBJECTIVETo establish a modified method for microculturing whole human blood for cytogenetic analysis.

METHODSA novel tube rack was designed to overcome the drawbacks of directly culturing the cells within centrifuge tubes. The fractions of human plasma, human serum and two commercial fetal bovine sera were analyzed with 15% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The influence of adding 0%, 5%, 10%, 15%, 20%, 25% and 30% autologous plasma to the culture on lymphocyte transformation rate and mitotic index (MI) was examined.

RESULTSThe SDS-PAGE analysis showed a significant difference between commercial fetal bovine sera, and that the components of human plasma were similar to those of fetal bovine serum. The value of MI in lymphocyte was evidently increased along with addition of autologous plasma. However, this has exerted no significant effect on the transformation rate. With the addition of 10% autologous plasma, the MI value has become much higher than the conventional method.

CONCLUSIONA modified method was established by application of a novel tube inclined rack and optimization of whole blood inoculation. This method is easier and cheaper, and is suitable for application in clinical practice.

Adolescent ; Adult ; Cell Culture Techniques ; methods ; Cytogenetics ; Female ; Humans ; Lymphocytes ; ultrastructure ; Male ; Mitotic Index