Objective To assess the impact of freeze-drying and dry heat virus inactivation processes on the activity ofα2-macroglobulin(A2M)derived from human plasma Cohn fraction Ⅳ.Methods A2M derived from human plasma Cohn fraction Ⅳ was prepared and subjected to programmed freeze-drying with dry heat virus inactivation.The lyophilized products were evaluated for their appearance,water content,and validation of the viral inactivation process.The bioactivity of the products before and after lyophilization as well as before and after dry heat inactivation was determined via trypsin inhibition,and the comparisons were studied.Results The appearance of the lyophilized product was fluffy,and the water content was(5.83±0.45)％.The specific activities of the samples before and after lyophilization were(10.199±0.137)and(10.033±0.201)μg/mg,respectively,with no statistically significantdifference between the two groups(P＞0.05).The viral inactivation of the samples was carried out by using dry heat inactivation conditions at 100 ℃ for 30 min.After inactivation,the reduction was ≥5.125 LgTCID50/0.1 mL in Pseudorabies virus(PRV)titers,≥4.500 LgTCID50/0.1 mL in Sindbis virus(SinV)titers,≥6.375 LgTCID50/0.1 mL in encephalomyocarditis virus(EMCV)titers,and≥4.500 LgTCID50/0.1 mL in porcine parvovirus(PPV)titers.The specific activities of the samples before and after dry heat were(9.921±0.292)and(10.091±0.278)μ g/mg,respectively,with no statistically significant difference between the two groups.Conclusion A2M derived from human plasma Cohn fraction Ⅳ,when subjected to freeze-drying followed by dry heat inactivation at 100 ℃ for 30 minutes,can effectively inactivate viruses without altering the biological activity of the product.

α2-Macroglobulin(A2M)is a high-abundance plasma protein with a molecular weight of 720×103,containing 1451 residues and 11 domains,and was isolated and identified for the first time in 1946.The capture and inhibitory effect on proteases is the classical biological function of A2M.However,A2M can also interact with membrane receptors,cytokines,and growth factors,and act as a molecular chaperone to affect extracellular protein homeostasis so that it is involved in a wide range of physiological and pathological processes such as immunity,inflammation,and degeneration.This article reviews the structural characteristics of A2M,the reported molecular targets,the mechanism of action,and its biological effects in the hope of providing a new line of thought for the functional exploration and clinical applications of A2M.

Objective To investigate the experience of intrauterine device(IUD)removal in patients with cervical cancer after radiotherapy.Methods A total of 23 patients with cervical cancer after radiotherapy underwent abdominal ultrasound guided hysteroscopic removal of IUD in our department from January 2020 to December 2022.For vaginal and/or cervical adhesions,blunt separation of adhesions was performed by using hysteroscope head or curved forceps under abdominal ultrasound guidance.If it was difficult to separate the adhesions for hysteroscope head entering the uterine cavity,a probe was inserted into the uterine cavity under ultrasound guidance,and the cervical canal was gradually expanded to 6-caliber dilation rod.Then the hysteroscope was inserted again into the uterine cavity.For obvious cervical atrophy that was tough in which cervical forceps could not be used to clamp the cervix,a 1-0 absorbable suture line was used to suture the anterior and/or posterior lips of the atrophied cervix at the top of the vagina,with an assistant firmly pulling and fixing the cervix.Results There were 4 cases of vaginal partial adhesions and cervical contracture,10 cases of cervical contracture,and remaining 9 cases having no vaginal adhesions and cervical contracture.Under hysteroscopy,there were 3 cases of cervical adhesions,2 cases of endometrial polyps,1 case of submucosal uterine fibroids,2 cases of uterine abscess,2 cases of incarcerated IUD,and remaining 13 cases having normal uterine cavity morphology.All the 23 cases of IUD were successfully removed by using abdominal ultrasound guided hysteroscopy(circular shaped in 12 cases,uterine shaped in 6 cases,V-shaped in 2 cases,Y-shaped in 1 case,T-shaped in 1 case,and umbrella shaped in 1 case).The surgical time was(19.2±10.9)min,and there were no complications such as false passage formation,uterine perforation,organ damage,massive vaginal bleeding,transurethral resection of the prostate syndrome,infection,embolism,or shock.The 23 cases were followed up for 2-24 months postoperatively,with a median of 12 months.One case continued concurrent radiotherapy and chemotherapy,3 cases continued post-loading radiotherapy,1 case continued chemotherapy,and 2 cases received targeted treatment(distant metastasis).The remaining 16 cases recovered well in regular reviews without complications such as abdominal pain,fever,or vaginal bleeding.Conclusions If the size and location of the cancer lesion do not affect the removal of IUD after radiotherapy for cervical cancer,it should be removed as soon as possible.The application of abdominal ultrasound guided hysteroscopy in IUD removal in patients with cervical cancer after radiotherapy is safe and feasible to a certain extent.

Objective     To analyze the clinical efficacy and survival outcome of totally thoracoscopic redo mitral valve replacement and evaluate its efficiency and safety. Methods     The clinical data of patients with totally thoracoscopic redo mitral valve replacement in Guangdong Provincial People’s Hospital between 2013 and 2019 were retrospectively analyzed. Survival analysis was performed using the Kaplan-Meier method. Univariate and multivariate Cox regression analyses were used to determine the risk factors for postoperative death. Results     There were 48 patients including 29 females and 19 males with a median age of 53 (44, 66) years. All the procedures were performed successfully with no conversion to median sternotomy. A total of 15, 10 and 23 patients received surgeries under non-beating heart, beating heart and ventricular fibrillation, respectively. The in-hospital mortality rate was 6.25% (3/48), and the incidence of early postoperative complications was 18.75% (9/48). Thirty-five (72.92%) patients had their tracheal intubation removed within 24 hours after the operation. The 1- and 6-year survival rates were 89.50% (95%CI 81.30%-98.70%) and 82.90%(95%CI 71.50%-96.20%), respectively. Age>65 years was an independent risk factor for postoperative death (P=0.04). Conclusion     Totally thoracoscopic redo mitral valve replacement is safe and reliable, with advantages of rapid recovery, reducing blood transfusion rate, reducing postoperative complications and acceptable long-term survival rate. It is worthy of being widely popularized in the clinic.

【Objective】 To establish and evaluate a fluorescent antibody to membrane antigen (FAMA) method for detecting antibodies against varicella-zoster virus (VZV) based on Vero E6 cells. 【Methods】 Based on the adapted VZV-Oka-E6 strain that VZV-Oka live attenuated varicella vaccine strain grew in Vero E6 cells, Vero E6 cells were infected with VZV-Oka-E6 of three different doses (10^4.65, 10^4.95 and 10^5.25 TCID50), and the cytopathic effect was observed under a microscope to determine the optimal infection dose of VZV-Oka-E6 strain. Then the detectable sensitivity of the infected cell antigen slides prepared after fixing the infected cells with different fixatives was compared to determine the optimal fixative. As a result, the FAMA method based on Vero E6 cells for the determination of neutralizing anti-VZV has been developed. The established FAMA assay was used to detect the international standard for varicella zoster immunoglobulin with different titers to determine the sensitivity of the assay. The specificity of the assay was evaluated by detecting specific antibodies against human herpes simplex virus (HSV) type 1 and type 2. The neutralizing anti-VZV antibodies of the international standard for varicella zoster immunoglobulin were detected using VZV-infected cell antigen slides prepared in the same batch and four different batches, respectively, to determine the intra-assay repeatability and inter-assay repeatability. The international standard for varicella zoster immunoglobulin with three known titers was detected to evaluate the relative accuracy of this assay. The anti-VZV titers in 20 apheresis plasma samples were determined with the newly established FAMA test and ELISA test, respectively, and the detection results of the two methods were compared using Spearman’s correlation test. 【Results】 The optimal infection dose of the VZV-Oka-E6 strain in FAMA assay was determined to be 10^5.25 TCID50, and acetone precooled at -20℃ was used as the fixative. The FAMA test has a high sensitivity with a detecting limit of 31.25 mIU/mL for neutralizing anti-VZV titers. The negative result was observed when detecting herpes simplex virus (HSV) type 1 and 2 specific antibodies. The international standard for varicella zoster immunoglobulin was detected by VZV infected cell antigen slides prepared in the same batch and 4 different batches, with the coefficient of variation being 29.95% and 26.71%, respectively. The detection value of the international standard for varicella zoster immunoglobulin with three different titer levels was consistent with their theoretical value. The correlation coefficient of the detection results of 20 apheresis plasma samples by the FAMA test and ELISA test was 0.268. 【Conclusion】 The VZV FAMA assay has good sensitivity, specificity, repeatability, and relative accuracy in detecting neutralizing anti-VZV titers. It can be applied for detecting neutralizing anti-VZV titers in apheresis plasma samples as well as the varicella-zoster immunoglobulin (VZIG) preparations.

Objective: To study the rapid antidepressant effects of cannabioiol(CBD) on depression-like mice and its possible mechanism. Methods: Chronic restraint was used to establish a mouse depression model. The test mice were divided into 5 groups: normal control group, model group, positive control group, CBD low-dose group (25 mg/kg) and CBD high-dose group(50 mg/kg). The mice in each group were given intragastric administration one hour before the behavioral experiment. After the behavioral experiment, the hippocampus and prefrontal cortex specimens were collected, and brain-derived neurotrophic factor(BDNF), postsynaptic density protein 95(PSD-95), synaptophysin(SYP) and target of rapamycin(mTOR) were tested by ELISA. Results: Compared with the model group, the central area activity distance percentage, the central area activity time percentage and total distance increased in the open field experiment in the CBD low-dose group. Compared with the model group, the percentage of immobility in the forced swimming experiment in the low-dose CBD group decreased. The ELISA test results showed that CBD could rapidly increase the concentration of BDNF and PSD-95 in the prefrontal cortex, as well as the concentration of SYP and mTOR in the hippocampus and prefrontal cortex. Conclusion CBD can rapidly improve the behavioral performance of depression-like mice, and rapidly up-regulate the level of BDNF and synaptic protein in the hippocampus or prefrontal cortex.

Objective To assess the quality of reports of clinical randomized controlled trials (RCTs) published in Chinese Journal of Dermatology from 2007 to 2016,and to provide a reference for standardization of clinical paper writing.Methods Based on the consolidated standards of reporting trials (CONSORT) 2010 statement,an evaluation form was designed and used to assess the quality of the clinical RCT articles published in Chinese Journal of Dermatology from 2007 to 2016.Results A total of 94 RCT articles were enrolled,including 45 articles from 2007 to 2011,and 49 articles from 2012 to 2016.Among these articles,the writing of introduction and discussion parts was relatively standardized.In the method and result parts,the proportions of articles correctly reporting blinding (23 articles,24.47％),sample size (0 article),primary and secondary outcome measures (21 articles,22.34％),participant flow (0 article),loss to follow-up (32 articles,34.04％) and compliance (13 articles,13.83％) were low,while the proportions of those correctly reporting inclusion and exclusion criteria,intervention measures,statistical methods,starting and ending time of follow-up and baseline data were all over 80％.Conclusion Most contents of the RCT articles published in Chinese Journal of Dermatology are standardized and clear,but the reporting of blinding,compliance,sample size,participant flow and so on,is insufficient,and close attention should be paid to these items.

Objective To investigate the mechanism by which hydrogen(H2) helps prevent acute lung injury induced by hyperoxia (HALI) in rats.Methods Thirty male Sprague-Dawley rats were randomly divided into three groups: control group, HALI group and H2 group, with 10 rats in each group.The control group was exposed to air at atmospheric pressure.Rats in HALI and H2 groups were exposed continuously to pure oxygen (100%O2) for 60 hours and during this period, 10 ml/kg of normal saline or H2-saturated normal saline was given every 12 hours by intraperitoneal injection to the HALI and H2 groups, respectively.After treatment, the arterial partial pressure of oxygen was examined and histopathological examination was conducted in each group.Then,RT-qPCR and Western blotting were performed to measure the transcriptional level and protein expression of heme oxygenase 1 (human heme oxygenase 1, HO-1) in rat lung tissue.Results Compared with the HALI group, H2 group showed significantly decreased severity of lung injury and a marked increase in the arterial oxygen saturation.Besides, H2 treatment induced up-regulation of HO-1 mRNA and protein levels.Conclusion The findings suggest that HO-1 may play an important role in the protection against HALI by H2.

Objective To develop an effective process for isolating and purifying haptoglobin ( Hp) from Cohn fractionⅣby a new ion exchange chromatography and to preliminarily identify and analyze the product of each purification step . Methods The fraction was first diluted and impurities were adsorbed with Rivanol .Then, the supernatant was treated with 50%ammonium sulfate.Finally, the precipitate was redissolved , and Hp was purified further with Q Sepharose Fast Flow chromatography .Native-PAGE was used to measure the activity of the haptoglobin-bound hemoglobin , while SDS-PAGE analysis and immunoblot were used for identification of the target protein .Results After pretreatment , some of the impuri-ties were removed from the Cohn fraction Ⅳ, and the target protein was enriched .In our case, the target protein was Hp and Hp2-2 was the main phenotype in the human plasma fraction Ⅳ.Target protein band and high purity were identified by SDS-PAGE.Immunoblot analysis further proved that this method could successfully isolate the target protein Hp , and the activity of 2.8 U/ml was measured by Native-PAGE method.Conclusion Haptoglobin is successfully isolated from human Cohn fractionⅣwith this method.The purification process is simple and suitable for scale-up production with a good prospect.

Plasma products are considered to be special medicinesderived from healthy human plasma .During 1980′s, events of transmission of human immunodeficiency virus through plasma products were frequently reported .Since then, ensuring the viral safety of plasma products has raised great concerns all over the world .So far, with decades of effort , most countries in the world have established rigorous systems with preventive measures to ensure the viral safety of plasma prod -ucts.These measures include control of source plasma , validated inactivation/removal of infectious agents , the adherence to current good manufacturing practices .Nevertheless , new infectious agents which may be threats to viral safety require continuous studies on appropriate countermeasures .