Objective To discuss the effect of quick freezer equipment and quick freezing time on the preparation of cryoprecipitate.Methods The cryoprecipitate,already prepared,was placed into the MBF21 freezer and minus 30℃ SANYO refrigerator for 30-minute and 50-minute storage.The activity of coagulation factor VⅧ,and the content of fibrinogen in different equipment were detected by automatic coagulation analyzer,in order to make sure whether it met the quality requirements.Results The content of fibrinogen in the cryoprecipitates from both of the equipment after 30-minute storage met the requirement,with the qualification rate of 100％.As to the activity determination of coagulation FⅧ,low temperature refrigerator showed a qualification rate of 62.5％,which was significantly lower than that of quick freezer with a qualification rate of 97.5％ (P＜0.01).The cryoprecipitates from both of the equipment after 50-minute storage,with part of fibrin precipitation,had a qualification rate of fibrinogen content lower than 50％ (P＞0.05).The qualification rate of quick freezer and low temperature refrigerator was 35％ and 12.5％,respectively,with significant difference.Conclusion Quick freezer can make the cryoprecipitate quick-frozen,which can ensure the quality.The coagulation factor VⅧ is unstable,whose activity decreased with the increase of temperature.We should try our best to shorten the time off the cold chain.

Nucleos(t)ide analogues (NAs) are effective inhibitors for HBV replication and have become the preferred antiviral regimen for most patients with chronic hepatitis B (CHB). HBeAg seroconversion is an important index used to evaluate the durability and efficacy of antiviral therapy in HBeAg-positive CHB patients. The search for biomarkers that can predict HBeAg clearance or seroconversion after NAs treatment plays an important role in the selection of antiviral drugs, the adjustment of treatment regimens, and the achievement of individualized treatment. This article reviews the value of related markers, including HBV DNA, HBV RNA, anti-HBc, and HBcrAg, in predicting HBeAg clearance or seroconversion in CHB patients treated with NAs.

Abstract: Objective　To analyze the clinical characteristics of children with lobar pneumonia and the distribution of pathogens in bronchoalveolar lavage fluid (BALF) collected from these patients, hence providing a scientific basis for their precise diagnosis and treatment. Methods　A total of 115 children diagnosed with lobar pneumonia from August 2019 to August 2022 at Suining Central Hospital were screened as the research subjects. The clinical manifestations and occurrence of complications in the patients were investigated. All the children underwent bronchoalveolar lavage after admission, and BALF samples were collected. Fluorescence quantitative PCR was adopted to detect and analyze the distribution and clinical characteristics of Streptococcus pneumoniae (SP) and other related pathogenic microorganisms in BALF specimens. Results Among the 115 pediatric patients with lobar pneumonia, the occurrence of manifestations or complications including involvement of ≥2 lung lobes, myocardial damage, pleural effusion, abnormal liver function, digestive system involvement, nervous system involvement, rash, renal function impairment, and lung atelectasis were observed in 46, 46, 39, 33, 18, 17, 11, 5, and 4 cases, respectively. The pathogen positivity rate in the BALF samples of the 115 patients was 87.0% (100/115), with 81 cases of single infection and 19 cases of mixed infection. A total of 121 strains of pathogens were isolated, including 83 strains of Mycoplasmal pneumonia (MP) (accounting for 68.6%) and SP(13.2%). The differences in the detection rates of HI, MP, RSV strains among different age groups were statistically significant (χ2=8.834, 19.454, 10.284, P<0.05), while the differences in the infection rates of SP, KP, CP, and ADV were not statistically significant (χ2=3.393, 2.67, 0.565, 0.097, P>0.05). The MP pneumonia group showed significantly higher incidence of complications such as pleural effusion, nervous system involvement, and abnormal liver function than the non-MP pneumonia group (χ2=3.925, 4.195, and 4.513, P<0.05). The highest pathogen detection rate was in winter, accounting for 33.91%. Conclusions MP is the most common pathogen in BALF of children with lobar pneumonia. There is variation in the pathogen detection rate among different age groups and seasons. Those with combined infections were more prone to complications, which is worthy of attention by clinicians.

Objective: To summarize and critically assess the evidence from randomized controlled trials (RCTs) of acupuncture in treating dry eye syndrome (DES) according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement and the Cochrane Collaboration recommendations. Methods: A search of PubMed, the Cochrane Central Register of Controlled Trials, ClinicalTrials.gov and Embase was made from their inception to August 2016, as well as Chinese, Japanese, and Korean databases. Two reviewers independently selected RCTs and assessed the methodological quality. Meta-analysis and the level of evidence were processed by RevMan 5.3 and the Grading of Recommendations Assessment, Development and Evaluation (GRADE) approach. Results: After selection, 8 trials were subjected to our systematic review. The methodological quality was low generally. The 3-10 weeks follow-up showed that acupuncture improved the tear film break-up time (BUT) (MD=1.33, 95%CI=1.01-1.66, 619 participants). The mean difference of Schirmer's test was 1.73 mm (95%CI=1.28-2.18, 618 participants) between the acupuncture group and the control group. The subjective variables exhibited no significant differences. Conclusion: The low methodological quality of the trials does not suggest drawing firm conclusions on the value of acupuncture therapy for DES. Acupuncture treatment may have some effects on the tear film BUT and Schirmer's test, but not on the subjective symptoms. Well-planned large-scale high-quality RCTs are needed to make it clear whether acupuncture is effective in treating DES.

Objective:To observe the therapeutic effect of warm needling moxibustion for erectile dysfunction (ED).
Methods:A total of 46 eligible ED patients were randomized into a treatment group (n=24) and a control group (n=22). The treatment group was intervened by warm needling moxibustion while the control group by conventional acupuncture treatment, for a total of 4 courses. The therapeutic effect was evaluated by the international index of erectile function 5 (IIEF-5).
Results:After 4 courses of treatment, the overall effective rate was 91.3% in the treatment group (23 completed), versus 75.0% in the control group (20 completed), with a significant difference (P<0.05). After the treatment, the change of IIEF-5 score in the treatment group was significantly different from that in the control group (P<0.05).
Conclusion:Warm needling moxibustion can produce a more significant efficacy than conventional acupuncture in the treatment of ED.

Objective To explore the differential level of hypermethylated SFRP2 gene in colorectal cancers andadjacent nontumorous tissues,to analyze the relation of SFRP2 gene promoter hypermethylation status and its clinicopathologic significance in colorectal carcinoma,and to study the relationship between the level of hypermethylated SFRP2 and the invasion and metastasis of colorectal carcinoma.Methods Real-time quantitative PCR technique was performed to analyze the level of hypermethylated SFRP2 gene promoter in colorectal cancers and adjacent nontumorous tissues taken from 30 colorectal cancer patients.The relation of the level of hypermethylated SFRP2 gene promoter and its clinicopathologic features of colorectal cancers was analyzed.Results SFRP2 gene promoter hypermethylation occurred in both the tumor tissues and the adjacent tissues.The level of SFRP2 gene promoter hypermethylation was significantly higher in the patients with TNM Ⅲ and Ⅳ (7.24 ± 1.13)than in patients with TNM Ⅰ and Ⅱ ( 5.92 ± 0.97 ) ( P ＜ 0.05 ).The level of SFRP2 gene promoter hypermethylation was significantly higher in the patients with lower differentiation(7.31 ± 1.11 ) than in patients with higher differentiation (6.23 ± 1.03) ( P ＜ 0.05 ).No significant association was found between the level of SFRP2 gene promoter hypermethylation and the status of gender,age.tumor location and tumor size ( P ＞ 0.05 ).Conclusions SFRP2 gene hypermethylation in tissues may be a useful objective parameter for the malignant level,invasion,metastasis,recurrence and prognosis of the colorectal cancer.

Objective This study is aimed to investigate the prognostic significance of ring sideroblast ( RS) in MDS( Myelodysplastic Sydrome ) and evaluate the correlation of RS and other prognostic index.Methods A total of 198 patients with MDS between March 2009 and December 2015 in Chinese PLA′s Gerneral hospital were chosen for this study .Based on the ratio of RS in nucleated red blood cell , patients were first separated into myelodysplastic syndrome without ring sideroblast (MDS RS-) group, RS≥15%, and myelodysplastic syndrome with ring sideroblast ( MDS RS +) group, RS <15%. Then, according to the proportion of blasts in bone marrow nucleated cells above 5%or below, patients were further divided into myelodysplastic syndrome with low blasts without ring sideroblast ( MDS-LB RS-) group, myelodysplastic syndrome with low blasts and ring sideroblast ( MDS-LB RS+) group, refractory anemia with excess blast without ring sideroblast ( RAEB RS-) group and refractory anemia with excess blast and ring sideroblast ( RAEB RS+) groupe.All patients had completed the morphological , genetics , molecular biology examination at dignosis, and followed up by phone.The results of the overall survival (OS) analysis have been presented in a Kaplan-Meier curve and cox regression model .Last, according to the percentage of RS in nucleated red blood cell , patients were separated into RS <5%groupe, 5%-15%group, 15%-40%group, RS≥40%group, and analyse their survival prognosis by statistical methods .Results Comparing to MDS RS-group, the morbidity age, WBC and PLT count were significantly higher [61 ±1.91 vs 52 ±1.37, t=-3.555, P<0.01, 3.82(0.47-323)vs 2.6(0.6-59.7), z=-4.014, P<0.01;139.5(7-608) vs 60(3-724), z =-3.988, P<0.01], bone marrow eythroid hyperplasia and gigantocyte were more obvious in MDS RS+group[χ2 =11.032, P<0.01, χ2 =5.165, P<0.05]; the percentage of GATA1 gene and abnormal rate of poor prognosis gene ( MLL, NRAS, WT1 ) , either mutation or high gene expression , were higher in MDS-LB RS+group than that in MDS-LB RS-( P<0.05 ); Contrasting with RAEB RS-group, the karyotype is worse in RAEB RS +group[χ2 =4.966, P<0.05];Comparing to 15%-40%group, the OS were poorer in RS≥40%;MDS RS+patients were more prone to adverse prognosis than MDS RS-patients.Conclusion Compared to MDS RS-group, MDS RS +patients had worse prognosis;RS maybe correlate to morbidity age , eythroid dysplasia and gene abnormality in affecting the survival prognosis of MDS.

Objective To reveal the mechanisms of immunological infertility,the method of coimmunoprecipitation(CO-IP) and liquid chromatogram mass/mass (LC-MS/MS) was used to screen sperm membrane proteins which interacting with antisperm antibodies (ASA).Methods This study was designed as a case-control.The disease group including 56 serum samples from 521 cryptogenic infertile patients were screened ASA positive by ELISA and conformed with mixed antiglobulin reaction(MAR).The controls were 31 serum samples which ASA is negative and already possessed healthy offspring.All subjects were enrolled from September 2015 to December 2015 in China PLA General Hospital.Spermatozoa samples from 48donors with normal sperm parameters were from January 2016 to April 2016 in China PLA General Hospital.The purified human sperm membrane proteins were then mixed with serum from disease group (positive for ASA) and control group (not containing ASA).The binding proteins of antisperm antibodies were enriched using CO-IP assay.The immunoprecipitates were separated on sodium dodecyl sulfate-polyactylamide gel (SDS-PAGE),then the binding proteins were cut from the gel and analyzed by LC-MS/MS after the enzymolysis.These proteins could bc idcntified as definition,biological function (s) and subccllular localization with Uniprot database.Results The serum samples from infertile persons (39 females and 17males) were screened ASA positive by ELISA and conformed with MAR.The healthy controls (17 females and 14 males) were ASA-negative in ELISA.Forty proteins that interact with ASA were obtained from the study and these could be divided into three groups:11 antigens detected by control serum samples only,14antigens recognised by both infertile patients and control sera,and 15 antigens specific for patients with ASA.These 15 proteins are Sperm Cation channcl protein 1,Sperm Cation channel protein 3,Sperm Cation channel protein 4,Sperm associated antigen 9,Apolipoprotein A-I,Dynein heavy chain 14,Cylicin-2,Izumo sperm-egg fusion protein 4,Thioredoxin domain-containing protein 2,IQ domain-containing protein H,IQ domain-containing protein F1,Spermatogenesis-associated protein 5,Spermatogenesis-associated protein 5-like protein 1,Sperm acrosome membrane-associated protein 1,E3 ubiquitin-protein ligase RNF 114.Conclusion Fifteen proteins discovered with CO-IP technology and LC-MS/MS analysis could be referred as male immunoinfertility-related antigens and they may hold the great importance in revealing the secret of immunological infertility.

Hepatitis C virus (HCV) infection is a global public health issue.At present,pegylated interferon (IFN) combined with ribavirin is the major therapeutic regimen for anti-HCV treatment in China,but this regimen cannot achieve ideal sustained virologic response.Direct-acting antivirals (DAA) have achieved marked clinical effects in the treatment of patients with hepatitis C.This article briefly describes the change in immune status after anti-HCV treatment from the three aspects of innate immunity,adaptive immunity,and cytokines/ chemokines.The analysis shows that HCV patients have complex immunoregulation,and the results vary from one study to another.In general,the expression of NK cell surface receptor and its function tend to recover and may even recover to a normal state.IFN treatment cannot restore the function of virus-specific CD8 + T lymphocytes,while the change in cytotoxic T lymphocytes after DAA treatment has not been clarified.Further studies are needed to elucidate the long-term effect of DAA on cvtokines and chemokines.

【Objective】 To investigate the status of occult hepatitis B virus infection(OBI) among blood donors in Quzhou, Zhejiang, and to analyze the mutation of S region in blood donors with anti-HBc+ alone and non anti-HBc+ alone. 【Methods】 The OBI samples were screened by ELISA and NAT; the HBV DNA was amplified and sequenced; 20 anti-HBc+ alone and 25 not anti-HBc+ alone samples were obtained. 【Results】 The detection rate of OBI in Quzhou was 0.10%(155/161 045), and the positive rate of anti-HBc was 74.19%(115/155). The detection rate of OBI increased with the age of blood donors(P<0.05), but was not related to gender. The positive rate of anti-HBc+ alone was 22.58%(35/155), and that of not anti-HBc alone was 51.61%(80/155). Among the 45 OBI sequencing samples, the proportion of B and C genotype was 73.33%(33/45) and 20.00%(9/45), respectively. The mutation sites of blood donors in the anti-HBc+ alone group were more than those in the not anti-HBc+ alone group, and the mutation rates of S114T and V168A on MHR were significantly different(P<0.05). 【Conclusion】 The genotype of OBI infection in Quzhou is mainly type B. The mutation sites of blood donors with anti-HBc+ alone are higher than those with not anti-HBc+ alone, which may be more suitable as one of the OBI screening indicators.