[Differences in Fungal Communities in Different Material Cellar Sediments and Their Correlation with Environmental Factors].

There are rich and diverse fungal communities in rainfall-cellar sediments. Fungi play a key role in the rainfall-cellar ecosystem as a bridge and link for material exchange between the rainfall-cellar ecosystem and the sediments. The changes in fungal community structure are usually closely related to the changes in environmental factors. The 16S rRNA gene Illumina MiSeq high-throughput sequencing technology was used to study the diversity and difference of fungal communities in the cellar sediments under two different catchment environments. The results revealed that the cellar sediments under the concrete catchment environment had higher diversity and richness of fungal communities than those under the loess land catchment environment. The dominant bacteria of the fungal communities under the two catchment environments were the same, namely Ascomycota, Basidiomycota, and Zygomycota, which constituted more than 90% of the abundance of the bacteria; however, the former had better homogeneity and stability. The indicator species based on LEfSe analysis demonstrated that Basidiobolales had the largest contribution to the diversity in the catchment environment of the loess land, and Mycosphaerella had the smallest contribution; Saccharomycetales contributed the most to the diversity in the concrete concentration environment, whereas Periconia contributed the least. The results of the co-occurrence network of the microbial community and environmental factors demonstrated that the positive relationship between fungi and environmental factors was stronger than the negative relationship. The research results have enhanced the understanding of the diversity of fungal communities in the cellar sediments and provided a reference for ensuring the drinking safety of rainwater harvesting cellar water for humans and livestock and improving the quality of cellar water.

[Oxidative Stress Status and Its Effect on Regulation of Hepcidin in Patients with HbHCS].

OBJECTIVE: To investigate the oxidative stress status and its effects on hepcidin in patients with hemoglobin H Constant Spring disease (HbH-CS). METHODS: A total of 35 patients were enrolled in the study, including 15 splenectomized cases and 20 non-splenectomized cases. 20 healthy volunteers were selected as controls. Serum superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH), oxidized glutathione (GSSG) levels, erythropoietin (EPO), serum free transferrin receptor (sFTR), growth differentiation factor 15 (GDF15) as well as the level of hepcidin were detected. Correlation analysis and multiple factor regression analysis were performed to investigate the factors affecting the iron metabolism and erythropoiesis. RESULTS: Compared with healthy control, the SOD and GSH levels in patients with HbHCS decreased, while MDA and GSSG levels increased. The levels of SOD, MDA, GSG and GSSG were not significantly different between the patients with splenectomy and those without splenectomy. Correlation analysis showed that inpatients with HbHCS, EPO, sFTR and GDF15 correlated negatively with SOD level and positively with MDA level. EPO and sFTR levels negatively correlated with Hepcidin. CONCLUSION: Excessive oxidative stress is present in patients with HbHCS, and hepcidin is inhibited by the upregulation of EPO and sFTR, and hence involved in iron overload in patients.

Human cytomegalovirus UL49 encodes an early, virion-associated protein essential for virus growth in human foreskin fibroblasts.

Despite recent results of deletion experiments showing that open reading frame (ORF) UL49 of human cytomegalovirus (HCMV) is essential, the expression, function and functional location of its encoded protein remain unknown. We generated an antibody specific for pUL49 to investigate the protein product encoded by the UL49 ORF and identified its function in HCMV-infected host foreskin fibroblasts. A bacterial artificial chromosome (BAC) of HCMV strain Towne (pRV-Towne) and the UL49-deleted mutant pRV-delUL49Towne were used to observe virus growth by plaque assay. Using a UL49-protein-binding antibody, we located pUL49 in the fibroblast cytoplasm. pUL49 exhibited expression kinetics resembling those of the class ß-2 proteins and was detected in the virion tegument. Following deletion of UL49 ORF, the virus failed to replicate, but it could be recovered by addition of pUL49 from pCDNA3.1 (+)-UL49. Our findings indicate that UL49 ORF is essential for HCMV replication in host foreskin fibroblasts.

[Comparisons of risk factors for low birth weight between Han and Uygur full-term infants].

OBJECTRIVE: To compare the differences in risk factors for low birth weight (LBW) between Han and Uygur full-term infants and to provide a basis for the prevention of LBW in newborn infants. METHODS: Eighty-seven full-term LBW infants (38 Hans and 49 Uygurs) between March 2013 and June 2014 were selected as the case group, and 186 full-term normal birth weight infants (92 Hans and 94 Uygurs) were selected as the control group. A questionnaire survey was performed to investigate the related factors for LBW. Multivariate logistic regression analysis was carried out to determine the risk factors for LBW. RESULTS: The birth weights in Uyghur LBW infants were lower than in Han ones (P<0.05). Multivariate logistic regression analysis showed that drinking (OR=2.472, P=0.015) and smoking (OR=2.323, P=0.007) by the father, pregnancy complications (OR=14.377, P<0.001), and times of pregnancy (OR=2.995, P=0.001) were the risk factors for LBW in Han infants, while drinking by the father (OR=1.968, P=0.007), times of pregnancy (OR=1.953, P=0.005), pregnancy complications (OR=10.283, P=0.002), and poor indoor environment (OR=1.367, P=0.027) were the risk factors for LBW in Uyghur infants. CONCLUSIONS: There are differences in physical growth between Han and Uygur LBW infants. Han and Uygur infants share the same traditional risk factors for LBW, such as father's harmful behaviors like drinking, times of pregnancy, and pregnancy complications, however, the indoor environment also plays a role in the occurrence of LBW in Uygur infants.

[Cell proteins that potentially interact with HBV polymerase were identified by co-immunoprecipitation-based LC-MS/MS identification and IPA].

Hepatitis B virus (HBV) is a major cause of chronic liver disease, and frequently results in hepatitis, cirrhosis, and ultimately hepatocellular carcinoma. HBV polymerase (Pol) is an essential viral protein that is important for HBV replication and might be involved in the development of hepatocellular carcinoma. Protein-protein interactions appears to be crucial for its role. The aim of this study was to screen and identify the proteins that interact with Pol using a co-immunoprecipitation-based LC-MS/MS identification technique. The HBV Pol gene was amplified by polymerase chain reaction (PCR) and cloned into pCDNA3.1(+). The recombinant plasmid pCDNA3. 1(+)-Pol-flag was transfected into HeLa cells. Liquid chromatography and tandem mass spectrometry (LC-MS/MS) identified 45 proteins that co-immunoprecipitated with flag-tagged HBV Pol. Eleven of these have previously been reported as proteins that interact with HBV Pol. A proof-of-concept-based Ingenuity Pathway Analysis (IPA, www.ingenuity.com) was used to characterize the functions and pathways of these 45 identified proteins and HBV Pol. Among these proteins, four proteins may play a role in three major molecular cellular networks, and are therefore worthy of further investigation.

Pulmonary hypertension risk in patients with hemoglobin h disease: low incidence and absence of correlation with splenectomy.

Pulmonary hypertension (PHT) is a common complication for patients with ß thalassemia intermediate (TI), especially splenectomized patients. However, the frequency and risk factors of PHT in patients with hemoglobin H (HbH) disease is unknown. The purpose of this study was to identify the prevalence of PHT risk manifested as tricuspid regurgitant jet velocity (TRV) ≥2.5 m/s in patients with HbH disease and its correlation with splenectomy. One hundred and ninety-eight patients with HbH disease who visited the 303rd Hospital of the People's Liberation Army (Nanning, China) were investigated. Thirteen subjects (6.5%) were diagnosed as having a risk of PHT. Regression analyses showed that the prevalence of PHT risk was correlated only with age (r = 0.195, p = 0.006) and not with splenectomy. The risk of PHT in patients older than 35 years was 5.7 times (range 1.8-18.6) greater than that for patients younger than 35 years. For splenectomized patients compared to those with HbH disease, patients with TI had a higher frequency of PHT risk, higher nucleated red blood cell counts (46.03 ± 41.11 × 10(9)/l vs. 0.18 ± 1.19 × 10(9)/l, p < 0.001) and a higher platelet counts (837.6 ± 178.9 × 10(9)/l vs. 506.7 ± 146.2 × 10(9)/l, p < 0.001). PHT risk is low in patients with HbH disease and does not correlate with splenectomy. Patients older than 35 years should be monitored regularly.

[BRPF2, a novel transcript of BRPF1 and their expression in mice tissues].

In order to identify a novel transcript of BRPF1 (BRPF2), a clone separated from mouse cDNA library was sequenced and submitted to GenBank. The expressions of BRPF1 and BRPF2 in different mice tissues were detected using RT-PCR and Northern blotting assays. The preliminary protein functions and conservative domains were analyzed by bioinformatic methods. The results indicated that BRPF2 was a novel transcript of BRPF1. Both BRPF1 and BRPF2 transcripts could be detected in most mice tissues, including liver, embryo, epididymis, testis, ovary and muscle. However, only BRPF2 transcript could be detected in the spleen. BRPF1 mRNA encoded 1 246 aa and the predicted molecular mass was 140 kDa, while BRPF2 encoded 442 aa, partly owing to the absence of a new stop codon. The results of CDD analysis suggested that BRPF2 lost the bromodomain and the PWWP domain compared to BRPF1. Because the bromodomain and the PWWP domain are the critical structures of BRPF1 to interact with histones and recruit the other transcriptional factors, BRPF2 (without these two critical domains) may serve as a negative regulatory factor of BRPF1 and be involved in the chromosome remodeling and transcriptional regulating.

Construction and expression of a recombinant eukaryotic expression plasmid containing the preS1-preS2-S genes of hepatitis B virus and the granulocyte-macrophage colony stimulating factor gene: A study of its immunomodulatory effects.

A total of 10-20% of the population remains unresponsive or weakly responsive to hepatitis B vaccine, which is composed of hepatitis B surface antigen HBsAg (S protein). Therefore, it is necessary to develop a hepatitis B vaccine with a better penetrating and responsive rate. In the present study, a plasmid pVAX1-L-GM was constructed and its immunomodulatory effect of as hepatitis B virus (HBV) DNA vaccine was analyzed through the immunization of BALB/c mice. Immune responses were measured after immunization by anti-HBsAg, proliferation of splenocytes, the number of CD4+ and CD8+ molecules, CTL cytotoxicity, cytokines of IFN-γ and IL-2 secretion assays. Following the immunization, mice in the pVAX1-L-GM group produced antibody 2 weeks earlier compared to the control plasmid pVAX1 and pVAX1HBsAg groups and antibody levels showed significant differences. Enhanced HBsAg-specific splenocyte proliferation as well as specific cytotoxic activities of splenic CTLs were also detected. Furthermore, pVAX1-L-GM plasmid increased the number of CD4+ and CD8+ molecules on the surface of the spleen T cell and the level of IFN-γ, IL-2 secretion. pVAX1-L-GM induced a specific immune response in mice and enhanced the immune effect. Thus, a foundation was laid for developing immunogenicity of a better prevention and treatment of HBV via a hepatitis B vaccine.

[The effects of human cytomegalovirus infection on college students' neurobehaviors].

OBJECTIVE: To explore relevant between human cytomegalovirus (HCMV) infection and college students' neurobehaviors. METHODS: 87 college students were enlisted. They were tested with Bole. Neurobehavioral evaluation system (B. NES), and HCMV IgG antibody was detected after separation of serum. We analyzed the test results of B. NES by SPSS software. RESULTS: 76 college students were infected by HCMV in the past and 11 college students were not infected. The infected group scored 8.89 +/- 6.60 in depression aspect of emotion state test, while control group got 15.73 +/- 9.00. There was Significant difference between infection group and control (P < 0.05). There were no significant differences in other aspects of emotion states, study and memory, perception and mental movement (P > 0.05). CONCLUSION: HCMV infection is associated with depression status.

[Identification of ubiquitously expressed transcript as the potential interactor of hepatitis B virus polymerase].

OBJECTIVE: To investigate the function of hepatitis B virus polymerase (HBV Pol) in the viral life cycle by screening the proteins interacting with HBV polymerase. METHODS: The HBV Pol gene was constructed into the pGBKT7 vector. GAL4 yeast two-hybrid system was used to screen the human liver cDNA library to obtain proteins which interacted with HBV Pol. GST-pull down assay was applied to confirm the protein interactions. RESULTS: Ubiquitously expressed transcript (UXT) was selected by the yeast two-hybrid system. GST-pull down assay confirmed the in vitro interaction between HBV Pol and UXT. CONCLUSIONS: UXT is a potential interactor of HBV Pol, and this protein interaction may provide clues of the function of HBV Pol in HBV life cycle.

Treatment and complications of thalassemia major in Guangxi, Southern China.

BACKGROUND: ß-Thalassemia is extremely prevalent in Guangxi province, Southern China. However, little is known about the treatment and complications of patients with thalassemia major (TM) in Guangxi. The first thalassemia center in China was opened in Guangxi in 2003. Since that time, more than 400 patients have been enrolled. PROCEDURE: From December 2009 to February 2010, data was collected from TM patients visiting the thalassemia center including the circumstances of diagnosis, biological and clinical data, markers of iron overload and treatment. RESULTS: Data on 231 patients (median age, 5 years; range, 5 months to 21 years) were recorded. Only 44.6% of patients maintained their hemoglobin levels >9.0 g/dl. In 186 patients with ferritin levels >1,000 ng/ml, an iron chelator was used regularly in 44.6%, irregularly in 26.9%, and was not used in 28.5%. The mean serum ferritin level was 3,143 ng/ml and levels increased with age. Height and weight retardation were found in 48.3% and 11.1% patients, respectively. Compared to patients treated outside of the center, patients completing treatment in the thalassemia center had a higher hemoglobin level before transfusion, higher height and weight SD score, and less splenomegaly, but a similar ratio of regular or irregular iron chelation. Six (18.2%) of 33 patients >10 years of age (14.3 ± 2.8 years; range, 11-19 years) were diagnosed as hypothyroid. CONCLUSIONS: Although survival status of patients with TM in Guangxi has improved since the opening of the thalassemia center, TM complications remain high and with an early onset.

A case report of concomitant paroxysmal nocturnal hemoglobinuria and heterozygous ß-thalassemia.

Paroxysmal nocturnal hemoglobinuria (PNH) is a rare clonal blood disorder that presents chronic intravascular hemolysis. PNH concomitant with inherited hemolytic anemia has been rarely reported. Here, we report an interesting PNH patient who was misdiagnosed with iron deficiency anemia due to concomitant heterozygous ß-thalassemia. The patient experienced dizziness, fatigue, and restricted physical activity for the previous 3 years. Thalassemia gene analysis revealed heterozygous ß-thalassemia. Iron staining of the bone marrow demonstrated the absence of stainable iron and sideroblasts. The patient was diagnosed with iron deficiency anemia. Iron supplementation treatment was performed, but the anemia remained unresolved. The patient became transfusion dependent 1 year later and was admitted to our hospital in March 2010. Flow cytometry of the patient's peripheral blood demonstrated that 7.9% and 11.9% of the erythrocytes were CD59 and CD55 deficient, respectively. The patient was finally diagnosed with concomitant PNH and heterozygous ß-thalassemia.

Hemoglobin H disease in Guangxi province, Southern China: clinical review of 357 patients.

The clinical characteristics of 357 patients with hemoglobin H (HbH) disease from the Guangxi province of Southern China were studied. One hundred and ninety-one (53.3%) patients were diagnosed with HbH-Constant Spring, 19 were diagnosed with HbH Westmead. Ten patients were shown to have coinherited HbH-Constant Spring/QS with a ß-thalassemia mutation. Coinheritance of the ß-thalassemia gene does not alleviate anemia (8.2 ± 2.3 vs. 7.6 ± 1.7 g/dl, p = 0.276), or influence age at diagnosis (20.2 ± 19.6 vs. 12.9 ± 11.0 years, p = 0.276). Ferritin levels were significantly higher in the group of patients with the nondeletional form of the disease (475 ± 719 vs. 249 ± 264 ng/ml, p = 0.005).

[Mutation analysis of the MMACHC gene in a pedigree with methylmalonic aciduria].

OBJECTIVE: To identify the mutation of the methylmalonic aciduria (cobalamin deficiency) CblC type, with homocystinuria (MMACHC) gene in a pedigree with methylmalonic aciduria. METHODS: The MMACHC gene mutation was detected using polymerase chain reaction (PCR) and DNA sequencing. The MMACHC gene of 50 healthy people was also sequenced as control. RESULTS: A new mutation of 146_154 del CCTTCCTGG was found in the patient and his father, and was absent in the controls. CONCLUSION: A new mutation (146_154 del CCTTCCTGG) in the MMACHC gene was detected in a Chinese family with methylmalonic aciduria.

[Studies of effect of prosaposin on cell proliferation, cell apoptosis and its possible molecular mechanism].

pcDNA3.1 in NIH3T3 and Psap-Myc in NIH3T3 cell strains were used as cell models in order to study the effect of prosaposin on cell proliferation, cell apoptosis and its possible molecular mechanism. MTT assay and Annexin V/PI apoptosis kit were used to detect the effect of prosaposin on cell proliferation and cell apoptosis induced by se-rum-starvation stress, respectively. Western blotting was conducted to detect the phosphorylative level of PI3K/Akt pathway, and real-time PCR was carried out to explore the expression of the genes regulated by PI3K/Akt pathway. Prosaposin pro-tein was proved to activate the PI3K/Akt signal pathway, upregulate the phosphorylative activity of Akt at Serine 473, downregulate the expression of P27(Kip1) gene, upregulate the expression of Cyclin D1 gene and then promote the G1/S tran-sition, and upregulate the expression of survival genes cIAP1 and cIAP2 and then prevent cell apoptosis. These findings suggest that the growth promotion and anti-apoptotic activity of prosaposin may be partly through the PI3K/Akt signal pathway and its downstream targeted genes.

Observations on after-effect duration of kidney-nourishing and marrow-replenishing therapy on 58 cases of Mediterranean anemia.

OBJECTIVE: To observe the after-effect duration of kidney-nourishing and marrow-replenishing therapy on Mediterranean anemia. METHODS: To observe the kidney-nourishing and marrow-replenishing therapy on 58 cases of Mediterranean anemia and the influence of various relative factors on the after-effect duration. RESULTS: The after-effect duration on 58 cases varied from 3-6 months, about 4 months on average, and was not influenced by sex, clinical types, genetic types, types of Mediterranean anemia and other factors. CONCLUSION: Kidney-nourishing and marrow-replenishing therapy used to treat Mediterranean anemia can not only produce good therapeutic effect during treatment but also keep after effect lasting for about 4 months, indicating that the therapy used to treat Mediterranean anemia has good clinical after effect.

[Identification and expression of human cytomegalovirus UL148 mRNA of low-passage clinical isolates in Guangzhou].

OBJECTIVE: To research the structure of the unique gene UL148 of the human cytomegalovirus (HCMV) clinical strain in Guangzhou, analyze the relation between its structure polymorphism and HCMV congenital infection, investigate the function of its expression products, and try to reveal the pathogenic mechanism of the gene in HCMV infection in newborns. METHODS: Urine samples were collected from 10 newborns with HCMV infection and inoculated on human fetal lung cells. The viral DNA was isolated, and UL148 gene fragment was amplified and identified. After TA clone and gene sequencing, total RNA of virus was extracted and the mRNA expression of UL148 gene was identified by using RT-PCR. RESULTS: The UL148 gene complete sequence was determined. A specific 582 bp length DNA fragment was amplified by RT-PCR, which confirmed the expression of UL148 gene. CONCLUSIONS: UL148 gene exists in the low-passage clinical HCMV isolate from Guangzhou. It is confirmed that open reading frame of UL148 has genetic structure with mRNA expression.

[Genomics innovative teaching pattern based upon amalgamation between modern educational technology and constructivism studying theory].

In order to overcome various malpractices in the traditional teaching methods, and also as part of the Guangdong province molecular biology perfect course project, some reforms were carried out to the teaching pattern of genomics. The reforms include using the foreign original teaching materials, bilingual teaching, as well as taking the constructivism-directed discussion teaching method and the multimedia computer-assisted instruction. To improve the scoring way and the laboratory course of the subject, we carried on a multiplex inspection systems and a self-designing experiments. Through the teaching reform on Genomics, we have gradually consummated the construction of molecular biology curriculum system.