Objective To explore the effect of methylene blue photochemistry(MB-P)viral inactivation treatment on the expression of microRNA(miRNA)in plasma exosomes,in order to provide a new reference for the quality control of MB-P virus inactivated plasma. Methods Whole blood samples of 11 healthy volunteers were collected from July 2021 to April2022. Fresh plasma from the same person was prepared into two parts,fresh frozen plasma(FFP)and MB-P virus inactivated plasma,respectively. The plasma exosomes were isolated by differential centrifugation,and identified by transmission electron microscopy(TEM)and nanoparticle tracking analysis(NTA). Then the expression profiles of miRNA were detected by microarray technique. Furthermore,four differentially expressed miRNA were verified by qRT-PCR,the target genes of differentially expressed miRNA were predicted by bioinformatics methods,and GO function enrichment analysis and KEGG pathway enrichment analysis were performed on the differentially expressed genes. Results The morphological characteristics and diameters of the extracted vesicles of the two groups were consistent with the characteristics of exosomes. Compared with the control group,there were 14 differentially expressed miRNA in plasma exosomes of MB-P group,of which the expression of six miRNA was up-regulated and eight miRNA was down-regulated. The results of qRT-PCR were generally consistent with the expression trend of microarray. The target genes of differentially expressed miRNA were mainly involved in DNA binding,ion binding,catalytic activity and other functions,and participated in a variety of biological processes such as nucleic acid metabolism,biosynthesis,and transcription regulation. In addition,significantly enriched functional pathways were closely related to viral infectious diseases,tumors,PI3K-Akt signaling pathway,MAPK signaling pathway and so on.Conclusion The expression of exosome miRNA in MB-P virus inactivated plasma was different from that in FFP. The plasma exosome miRNA may be used as a potential reference for the quality evaluation of MB-P virus inactivated plasma.

Objective To screen an effective method for removing free polysaccharides in polysaccharide-protein conjugate of pneumococcus type 9V(Pn9V),and to optimize the process and verify its stability,so as to use it in the preparation of multivalent conjugate vaccine against pneumococcus.Methods Pn9V-CRM_(197) conjugate was prepared,of which the free polysaccharides were removed by molecular sieve(Sepharose 4FF),ion exchange chromatography(DEAE Sepharose FF),hydrophobic chromatography(GP-Butyl)and ammonium sulfate(AS)salting out. The optimal method was selected by comparing the percentage of free polysaccharides,the recovery rate of polysaccharides and the recovery rate of proteins,and the determined method was optimized for the process. Three batches of conjugates were prepared continuously to verify the stability of the process.Results Both hydrophobic chromatography and ion exchange chromatography reduced the free polysaccharides in Pn9V-CRM197 conjugate to less than 5%. The recovery rates of polysaccharide and protein by hydrophobic chromatography were 34. 7% and 50. 1%,respectively,which were higher than those by ion exchange chromatography(16. 2% and 25. 7%). Hydrophobic chromatography was determined with the optimized process including 1. 1 mol/L AS as the loading buffer,water for injection used for elution,and the conjugate loading of 1. 81 mg protein/mL filler. The free polysaccharide removal rate,polysaccharide recovery rate and protein recovery rate of the conjugates prepared continuously in three batches were good,and the consistency among batches was high. The conjugates had good stability when stored at 2-8 ℃ for 6 months,and the percentage of free polysaccharides increased significantly(F = 5. 83 e~(-32),P = 0. 003)and was more than25% when stored at(25 ± 2)℃ for 6 months,indicating that high temperature storage was not suitable for the conjugates.Conclusion A hydrophobic chromatography process was established to effectively remove free polysaccharides from Pn9V polysaccharide-protein conjugate,which has good stability and provides a reference for the removal of free polysaccharides in other pneumococcal polysaccharide-protein conjugates.

Objective To analyze and discuss the clinical value of tuberculosis infection T cell ELISPOT test and tuberculosis antibody test used in bone and joint tuberculosis.Methods The patients with suspected bone and joint TB diagnosed and treated in Xishan people's Hospital of Wuxi City from 2012 to 2014 were selected as the objects of this study.They were taken randomly,which divided into T-SPOT.TB test group and tuberculosis antibody test groups.Each group had 162 per-sons.The patients of T-SPOT.TB test group were tested by tuberculosis infection T cell enzyme-linked immunosorbent spot test,tuberculosis antibody test group were taken tuberculosis antibody test,comparative analysis of the sensitivity and speci-ficity of two diagnostic methods were contrastively analyzed.Results The sensitivity and specificity of T-SPOT.TB test group were 93.43% and 84.00%.The sensitivity and specificity of tuberculosis antibody in the experimental group were 40.44% and 53.85%.There was statistical difference between two groups (χ2 =184.62,7.86,all P <0.05).The false posi-tive rate and the false negative rate of T-SPOT.TB test group were 16.00% and 6.57%.The false positive rate and the false negative rate of tuberculosis antibodies in the experimental group were 26.92% and 59.56%,respectively.There was statis-tical difference between two groups (χ2 =7.86,156.18,all P <0.05).In addition,the positive and negative predictive values of T-SPOT.TB test group were 96.97% and 70.00%.Positive and negative predictive values of tuberculosis antibody in the experimental group were 88.71% and 14.74%.There was statistical difference between two groups (χ2 = 20.27,73.06,all P <0.01).Conclusion Compared to experimental tuberculosis antibody,TB infection of T cells ELISPOT test had high sen-sitivity and specificity in the diagnosis of bone and joint tuberculosis.It’s worthy of spreading.

Objective To discuss the techniques of ureteroscopy in the diagnosis and treatment of ureteral diseases. Methods The authors retrospectively analyzed clinical data ot 92 patients with ureteral diseases undergoing the ureteroscopy. Results The success rate of ureteroscopy was 92 4% (85 of 92). Except for 2 cases with the complications of false urethra, no other complications were seen. Conclusions By the guidance of ureteral catheter, it is a safe and reliable approach to enter the ureteroscope directly or spirally by means of hand-controlled intermittent perfusion dilation. And more stones can be removed by intermittent low-flow perfusion and by the use of ureteral catheter or basket for stone fixation.

ObjectiveTo investigate the outcomes of mother a nd baby smaller than 37 gestational age caused by preeclampsia. Method sThe data for 122 preterm birth that was single pregnancy with preecla mpsia in our hospital from January 1993 to December 2002 were collected. The pri nciple of diagnosis and treatment, the outcomes were analyzed retrospectively. T he 122 pregnant women were divided into 3 groups according to their gestational age, more than 28 weeks and smaller than 31 +6 weeks was the first group (28 cases), more than 32 weeks and smaller than 33 +6 weeks was the second grou p (26 cases), more than 34 weeks and smaller than 36 +6 weeks was the thir d group (68 cases). The data were statisted with SAS software.Results The earlier the pregnancy-induced hypertension happened, the earlier the preeclampsia occured, the mean time was 14 03 d, 16 85 d,22 22 d, respect ively, (P0.05), the outcomes of mothers were good. Increasing the gestational age under the cond ition that the occurrene of comptications was not increased, 60 64% pregnant wo men accepted the treatment of improving the maturity of fetal lung. There were n o differences among the three groups at the morbidity of perinatal SGA, intracra nial hemorrhage (P 0.05) .ConclusionWe could improve the outcomes of pret erm birth caused by preeclampsia by enhancing the perinatal care, treating and t erminating pregnancy activey Increasing the gestational age, improving the matur ity of fetal lung would decrease the morbidity of severe RDS and improve the pe rinatal outcome effectively.

ObjectiveTo investigate the relationship between carotid atherosclerosis and progressing stroke.Methods103 patients with acute cerebral infarction within the first 24 hours from onset were tested with carotid color doppler ultrasonography during the first week from admission. Univariate and multiple logistic regression analyses were performed to examine the relationship between carotid intima media thickness (IMT), plaque structure, the degree of artery stenosis and the early deterioration of cerebral infarction.ResultsPrevalence of carotid plaque and thickness of IMT in patients with progressing stroke were higher than that in non progressing patients. In logistic regression, presence of carotid plaque was independent predictors of progressing stroke. Plaque size, extent of artery stenosis and IMT were not included in the regression model.ConclusionPlaque structure and its chemical feature may be the cause of progressing stroke.

OBJECTIVE: To compare the stability between imported branded and domestic entecavir active pharmaceutical ingredient(API) and among four Entecavir products (A for the original brand product and B-D for the domestic generic agents) and to provide references for clinical use. METHODS: The influencing factor tests and accelerated tests of the API and the four products were carried out. The stability of the imported and domestic API was studied. The stability of the product was studied by comparing the appearance, weight of tablets, API content, related substance and dissolution. RESULTS: The influence of illumination on the stability was the least, while high temperature and high humidity had the most significant influence. In the accelerated tests, product A was the most stable one while products B-D changed with varying degrees, which showed up in appearance, the API content, the related substances and the dissolution. CONCLUSION: The stability of the API from different sources appeared to be similar, while that of the four products showed differences. The stability of the original product was noticeably superior to the domestic generic agents. Copyright 2012 by the Chinese Pharmaceutical Association.

ATP1A3 gene mutations are associated with a wide spectrum of neurological symptoms. This article describes a case in children, which is caused by mutation of ATP1A3 gene. A seven years old boy presented with symptoms of fever-induced generalized dystonia, dysarthria, and paroxysmal weakness. Gene sequencing results showed that the ATP1A3 gene in the boy had heterozygous mutations of c.1838C>T. Combined with the clinical manifestations, gene mutations and literatures, the patient was diagnosed as a phenotypic overlap of an atypical form of rapid onset dystonia-parkinsonism and paroxysmal weakness. The results of genetic testing can contribute to the early diagnosis of patients with atypical and overlapping phenotypes in ATP1A3 spectrum disorder.

Maternal heart disease has emerged as one of the major diseases threatening pregnant women's safety and long-term cardiovascular health. It is worth noting that there is a large gap in pregnancy prognosis with various cardiovascular diseases. The American College of Obstetricians and Gynecologists (ACOG) has published a practice bulletin named "Pregnancy and Heart Disease" in April 2019, which addressed the prevalence, risk factors, and heart disease effects on pregnancy and the puerperium. It outlined the common types and tiered care recommendations for cardiovascular diseases during pregnancy and puerperium.