[Analysis of risk factors and prognosis of cytomegalovirus infection post umbilical cord blood stem cell transplantation in children with primary immunodeficiency diseases].

Objective: To investigate the risk factors and outcomes of cytomegalovirus (CMV) infection post umbilical cord blood stem cell transplantation (UCBT) in children with primary immunodeficiency diseases (PID). Methods: Clinical data of 143 PID children who received UCBT in the Children's Hospital of Fudan University from January 2015 to June 2020 were collected retrospectively. CMV-DNA in the plasma was surveilled once or twice a week within 100 days post-UCBT. According to the CMV-DNA test results, children were divided into the CMV-infected group and the CMV-uninfected group. The incidence and risk factors of CMV infection were analyzed. At 1-month post-UCBT, the absolute lymphocyte count, ratio of lymphocyte subsets and immunoglobulin levels were compared between those whose CMV infection developed 1-month later post-UCBT and those not. Mann-Whitney U test and chi-squared test were used for comparision between groups. Kaplan-Meier survival analysis was used to analyze the impact of CMV infection on survival. Results: Among 143 patients, there were 113 males and 30 females, with a age of 14 (8, 27) months at UCBT. Chronic granulomatosis disease (n=49), very-early-onset inflammatory bowel disease (n=43) and severe combined immunodefiency (n=29) were the three main kinds of PID. The rate of CMV infection was 21.7% (31/143), and the time of infection occurring was 44 (31, 49) days post-UCBT. The incidence of recurrent CMV infection was 4.2% (6/143) and refractory CMV infection was 4.9% (7/143).There was no significant difference in the first time CMV-DNA copy and peak CMV-DNA copy during treatment between the recurrent CMV infection group and the non-recurrent CMV infection group (32.8 (18.3, 63.1)×106 vs. 22.5 (13.2, 31.9)×106 copies/L, Z=-0.95, P=0.340;35.2 (20.2, 54.6)×106 vs. 28.4 (24.1, 53.5)×106copies/L, Z=-0.10, P=0.920), so were those between the refractory CMV infection group and non-refractory CMV infection group (21.8 (13.1, 32.2)×106 vs. 25.9 (14.2, 12.2)×106copies/L, Z=-1.04, P=0.299; 47.7 (27.9, 77.6)×106 vs. 27.7 (19.7,51.8)×106copies/L, Z=-1.49, P=0.137). The CMV-infected group accepted more reduced-intensity conditioning (RIC) regimen than the CMV-uninfected group (45.2% (14/31) vs. 25.0% (28/112), χ2=4.76, P<0.05). The rate of CMV-seropositive recipients and Ⅱ-Ⅳ acute graft versus host diseases (aGVHD) are significantly higher in the CMV-infected group than the CMV-uninfected group (100% (31/31) vs. 78.6% (88/112), 64.5% (20/31) vs. 26.8% (30/112), χ2=7.98,15.20, both P<0.05). The follow-up time was 31.6 (13.2, 45.9) months, CMV infection had no effect on overall survival (OS) rate (χ2=0.02, P=0.843). There was significant difference in the survival rate among three groups of refractory CMV infection, non-refractory CMV infection and the CMV-uninfected (4/7 vs.95.8% (23/24) vs. 86.6% (97/112), χ2=5.91, P=0.037), while there was no significant difference in the survival rate among three groups of recurrent CMV infection, non-recurrent CMV infection and the CMV-uninfected (5/6 vs. 88.0% (22/25) vs. 86.6% (97/112), χ2=0.43, P=0.896). Children who developed CMV infection after 30 days post-UCBT had lower absolute count and rate of CD4+ T cells and immunoglobulin G (IgG) level than those in the CMV-uninfected group (124.1 (81.5, 167.6) ×106 vs. 175.5 (108.3, 257.2) ×106/L, 0.240 (0.164, 0.404) vs. 0.376 (0.222, 0.469), 9.3 (6.2, 14.7) vs. 13.6 (10.7, 16.4) g/L, Z=-2.48, -2.12,-2.47, all P<0.05), but have higher rate of CD8+T cells than those in CMV-uninfected group (0.418 (0.281, 0.624) vs. 0.249 (0.154, 0.434), Z=-2.56, P=0.010). Conclusions: RIC regimen, grade Ⅱ-Ⅳ aGVHD and CMV-seropositive recipients are the main risk factors associated with CMV infection in PID patients post-UCBT. Survival rate of children with refractory CMV infection after UCBT is reduced. Immune reconstitution in children after UCBT should be regularly monitored, and frequency of CMV-DNA monitoring should be increased for children with delayed immune reconstitution.

[Research on the influence of coronavirus infection on maternal and infant health].

COVID-19 spreads with strong infectivity and triggered a public health crisis, home and abroad. SARS-CoV-2 has high pathogenic homology with SARS-CoV and MERS-CoV, and the three coronaviruses all belong to the Betacoronavirus family. Due to pregnant women's physical and psychological vulnerability, they are the susceptible and high-risk groups during the epidemic. This article will review the reports on adverse effects of maternal and fetal health during the SARS and MERS and COVID-19 epidemics to provide evidence for the clinical management and prevention and control of pregnant cases in SARS-CoV-2 infection.

Andrographolide and its derivatives are effective compounds for gastrointestinal protection: a review.

OBJECTIVE: Andrographolide and its derivatives have many functions, such as anti-infection, anti-tumor, neuroprotection, and immune regulation. However, the gastrointestinal protective effects, especially gastrointestinal tumors, and inflammation-related diseases of andrographolide and its derivatives have not been well summarized and discussed. In this review, we aimed to summarize and discuss the pharmacological effects and underlying mechanisms of andrographolide and its derivatives in gastrointestinal protection, with a view to revealing more possibilities of andrographolide and its derivatives in gastrointestinal diseases prevention therapy. MATERIALS AND METHODS: The data in this review are searched and selected from PubMed with the keywords: Andrographolide and Andrographolide derivatives, and relevant data with gastrointestinal protection are extracted and discussed. RESULTS: Andrographolide and its derivatives have prophylactic and therapeutic effects in gastrointestinal disorders such as GU, gastric cancer, colorectal cancer, and inflammatory bowel disease. CONCLUSIONS: Andrographolide and its derivatives are effective compounds for gastrointestinal protection.

[Epidemiological survey of mental disorders in the rural left behind elderly aged 60 years and older in Gansu].

Objective: To investigate the prevalence, demographic characteristics and social life function of mental disorders in the rural left behind elderly aged 60 years and older in Gansu. Methods: Between November 2017 and June 2018, a multi-stage stratified cluster sampling method was used to randomly select the rural left behind elderly aged 60 years and older in Gansu, and totally 6 000 elderly were enrolled. By using the extended general health questionnaire (GHQ-12) and the American Handbook for Diagnosis and Statistics of Mental Disorders (DSM-Ⅳ) Axis Ⅰ Disorders Formal Clinical Examination Patient Edition, all the included subjects were screened and diagnosed. Functional status was assessed by the Global Assessment Function scale (GAF). Statistical analysis of the prevalence of various mental illnesses, as well as the differences in the prevalence of different gender, marital status and age groups was performed. Results: Totally, 6 000 subjects completed the survey. The adjusted current prevalence of any mental disorder was 20.11% (95%CI 17.70%-22.85%). The six most prevalent specific disorders were major depressive disorder (9.20%), pain disorder (2.71%), mood disorder due to the body condition (2.08%), generalized anxiety disorder (1.99%), anxiety disorder not otherwise specified (1.15%) and dysthymic disorder (0.84%). The lifetime prevalence of mental disorders was 20.54% (95%CI 18.40%-23.39%). The overall current prevalence of mental disorders was higher in women (242.89‰) than in men (119.55‰), and the unmarried (248.37‰) was higher than those married (187.53‰). There was no significant difference in the prevalence of mental disorders among different age groups (P>0.05). The GAF score of mental disorders was 56±11, and 71.82% was moderate to severe functional impairment. Conclusions: The prevalence of mental disorders is high in rural left-behind population aged 60 years and over in Gansu Province. Major depression is a condition that deserves special attention.

MicroRNA-409 inhibits the proliferative ability of cervical carcinoma cells by regulating AKT.

OBJECTIVE: To observe the microRNA-409-3p expression in cervical carcinoma and its effect on the growth and proliferative ability of cervical carcinoma cells. PATIENTS AND METHODS: The microRNA-409-3p level in 62 cases of cervical carcinoma and 38 cases of normal cervical tissue was detected by qRT-PCR. The association between the microRNA-409-3p level and clinicopathological features of cervical carcinoma was investigated. Knockdown and overexpressed microRNA-409-3p in cervical carcinoma cells, HeLa and SiHa, were used to detect the cell cycle and the activity of cervical carcinoma cells. Subsequently, potentially target genes of microRNA-409-3p were predicted by bioinformatics website. Western Blot and luciferase assay were used to confirm their correlation. RESULTS: We observed a significant lower microRNA-409-3p level in cervical carcinoma tissues than that in normal cervical tissues. Significant correlation was found between the microRNA-409-3p level in patients with cervical carcinoma and the overall survival rate, tumor size and TNM stage (p<0.05), but correlations with age, pathological type and lymph node metastasis were not found (p>0.05). After silencing the expression of microRNA-409-3p in cervical carcinoma cells, the proliferative ability of cervical carcinoma cells was greatly promoted. In addition, microRNA-409-3p was targeting on protein kinase B (AKT) and had a negative correlation with AKT expression. CONCLUSIONS: The microRNA-409-3p level was lower in cervical carcinoma tissues, and was significantly affected the overall survival, tumor size and TNM staging in clinical patients. Low expression of microRNA-409-3p could promote the proliferative ability of cervical carcinoma cells through the target gene AKT.

Screening for genes that are differentially-expressed between gastric cancer cells and gastric tumor sphere cells using the gene chip technique.

The purpose of this study was to screen for genes that were differentially expressed between a human gastric carcinoma cell line (HGC-27) and their tumor spheres, using the gene chip technique. The HGC-27 cells and tumor sphere cells were cultured in vitro in a sterile environment. Total RNA was extracted from both samples and purified using a standard TRIzol reagent. Total RNA was then hybridized onto a GeneChip, according to the standard protocols provided by the manufacturers of the GeneChip IVT Express Kit. The resulting fluorescence signals were analyzed and displayed using the Cluster and Treeview software programs. Under the criteria for significant differential expression (≥2-fold difference), 610 up- and 1135 down-regulated genes were identified in tumor sphere cells, compared to HCG-27 cells. These genes were involved in cell growth, signal transduction, tumorigenesis, and many other functional aspects of tumor cells. In conclusion, a number of genes were differentially expressed in tumor sphere cells compared to HCG-27 cells. In addition, we identified a close correlation between tumor sphere cells and tumorigenesis.

Double activation improves rabbit freeze-thawed oocytes developmental potential.

OBJECTIVE: To investigate the effects of various activation methods on freeze-thawed rabbit oocytes developmental potential. METHODS: Rabbit oocytes were vitrified by cryoleafs and cryoprotected with ethylene glycol and propanediol. After thawing, the oocytes were fertilized by intracytoplasmic sperm injection (ICSI). Surviving oocytes after ICSI were divided into five groups at random. Group 1: Oocytes (n = 30) activated 1 h after ICSI by calcium ionomycin (I0634); Group 2: Oocytes (n = 26) activated by strontium chloride an hour after ICSI; Group 3: Oocytes (n = 33) activated by I0634 twice; Group 4: Oocytes (n = 28) were activated by strontium chloride twice; CONTROL GROUP: Inactivated oocytes (n = 39). Blastocysts derived from each group were transplanted to recipient rabbits. RESULTS: Rates of fertilization, cleavage and blastocyst formation of Group 3 were higher than those of Group 1 and Group 2 (81.8% vs 33.3% vs 53.8%, 54.5% vs 16.7% vs 26.9%, p < 0.05; 15.2% vs 3.3% vs 7.7%, p > 0.05). The rabbit transplanted with embryos derived from Group 3 became pregnant. Embryos derived from double activation could implant into endometrium. CONCLUSION: Double activation may increase freeze-thawed oocytes developmental potential. After activation, oocytes cleavage velocity may be faster than that of oocytes without activation.

Human umbilical cord blood serum in culture medium on oocyte maturation In vitro.

In vitro maturation (IVM) of immature oocytes for infertile patients is an attractive treatment. It can avoid side effects of ovarian stimulation with gonadotropins. However, at the present the successful results of IVM treatment are lower than conventional in vitro fertilization (IVF) treatment. The key issue may be the IVM medium for immature oocyte maturation. In the present study, we compared 20% (v/v) human follicular fluid (hFF) and 20% (v/v) human umbilical cord serum (hUS) as a supplement to IVM medium. A total of 47 patients with polycystic ovary syndrome (PCOS) underwent 47 IVM treatment cycles. Immature oocytes (349) collected from 32 patients were cultured in IVM medium supplemented with hFF, and immature oocytes (160) collected from 15 patients were culture in IVM medium supplemented with hUS. The results indicate that the final maturation rate of oocytes cultured in IVM medium supplemented with hUS (93.8%) is significantly higher than those cultured in IVM medium supplemented with hFF (77.1%). The percentage of high-quality embryos produced from IVM medium supplemented with hUS (50.0%) is significantly higher than IVM medium supplemented with hFF (23.8%). In addition, the results also indicate that the final maturation rate of oocytes is higher in IVM medium supplemented with hUS and the time course of oocyte maturation is hastened. Following transfer 6 out of 15 patients (40.0%) become pregnant when IVM medium was supplemented with hUS, and 7 out of 31 patients (22.6%) were pregnant when IVM medium was supplemented with hFF. These results suggest that IVM medium containing hUS appears to be a more effective means to stimulate in vitro oocyte maturation and is capable of achieving a promising clinical outcome.

[Alzheimer-like protein phosphatase deficiency leads to abnormal phosphorylation and accumulation of neurofilaments].

OBJECTIVE: To explore the effect of Alzheimer-like protein phosphatase deficiency on neurofilament phosphorylation. METHODS: Cell culture, light microscopy, immunocytochemistry and biochemistry techniques were used to make a phosphatase deficient cell model, to detect cell morphology, neurofilament phosphorylation and distribution, cell viability and activity. RESULTS: Non-phosphorylated neurofilament recognized by SMI32 was detected both in cell body and cell processes, it was extremely enriched in cell bodies; Phosphorylated neurofilament bound to SMI34 was mainly determined in cell processes and cell surface. After treatment with okadaic acid (OA), non-phosphorylation-dependent antibodies SMI32 staining was significantly decreased in the cell body, whereas phosphorylated neurofilament reacted with SMI34 was strikingly increased in immunocytochemistry and Western blot, and prominently accumulated to the same cell location. Accompanied with hyper-phosphorylation and accumulation of neurofilament, dose dependent cell toxicity was observed by okadaic acid treatment. CONCLUSION: Deficiency in protein phosphatase induces in neuroblastoma cell line, neurofilament phosphorylation and accumulation, which is involved in Alzheimer neurofibrillary degeneration.

[Comparative study of the expression and phosphorylation of neurofilament proteins of brain gray matter in Alzheimer's disease].

OBJECTIVE: To explore the role of abnormal modification of neurofilament proteins (NFPs) in the pathogenesis of Alzheimer disease (AD), and the characteristics of NFPs in distribution and solubility. METHOD: Ultra-centrifugation and Western blot were utilized to isolate and analyze NF subunits. Ultra-centrifugation and Western blot were utilized to observe the effect of abnormal modification of neurofilament proteins (NFPs) on the characteristics of NFPs in distribution and solubility in the pathogenesis of Alzheimer disease. RESULTS: In human brain gray matter, NF subunits were mainly detected in particular fraction. Compared with Huntington's disease (HD), the content of NF subunits was increased in AD brain gray matter and the increased level of NF-H and NF-M were mainly in abnormal hyper-phosphorylate form. CONCLUSION: Abnormal phosphorylation and accumulation of NFPs are involved in pathology of AD brain.

Microbial transformation of 2-hydroxy and 2-acetoxy ketones with Geotrichum sp.

Biotransformation of a series of o-, m- and p-substituted alpha-hydroxy- and alpha-acetoxyphenylethanones 1a-h and 9a-g with Geotrichum sp. led to the corresponding 1,2-diols 2 and/or monoacetates 10 in moderate to excellent enantiomeric excesses. Alpha-hydroxy- and alpha-acetoxyphenylethanones and their m- and p-derivatives gave preponderantly the S-configuration products while in the case of the o-derivatives R-alcohol was provided as the major enantiomer. The results of stereoselectivity were discussed.

Metabolism of n-VLDL and beta-VLDL in peritoneal macrophages from diabetic mice.

Peritoneal macrophages were harvested from both healthy mice and mice with diabetes induced by intravenous injection of streptozotocin. After incubation with normal plasma very low density lipoprotein (n-VLDL) from swine for 24 h, the content of triglyceride (TG) in macrophages from diabetic mice was higher than that in the controls, and it was the same with the concentration of free fatty acid (FFA) in medium. The basic concentration of cholesterol ester (ChE) in macrophages from diabetic mice was also higher than that of the controls. After incubation with beta-VLDL from hypercholesterolemic rabbit for 48 h, no alteration in ChE content was found in the cells. On the contrary, the content of ChE in the cells from the controls was accumulated by saturating. The levels of FFA in the medium of the diabetic group were still higher than those of the controls. These results demonstrated that the metabolic function of peritoneal macrophages in diabetic mice was altered. The uptake and degradation of n-VLDL and the endogenous cholesterol synthesis were increased, but the uptake of beta-VLDL was lower. In this paper, the possible mechanism and the relationship between atherosclerosis and diabetes are discussed.