Phylogenetic relationships and genetic differentiation of two Salamandrella species as revealed via COI gene from Northeastern China.

Due to traditional classification methods' limitations, some cryptic species remain undiscovered. To better explore the existence of the Schrenck salamander (Salamandrella tridactyla, a cryptic species of Siberian salamander S. keyserlingii) in China, we conducted a molecular phylogenetic analysis to confirm the taxonomic relationship among Salamandrella species and investigate genetic variation. We used complete sequences of the mitochondrial COI gene from 65 specimens collected across a wide range in Northeastern China. Thirty-five haplotypes were obtained from six populations. They showed medium-high haplotype diversity (Hd) and low nucleotide polymorphism (π). The phylogenetic tree and haplotype network analysis revealed that populations from Greater Khingan Ridge (Huma: HM) and Lesser Khingan Ridge (Tieli: TL) belong to S. keyserlingii, while populations from Changbai Mountain (Shangzhi-zhuziying: SZ, Shangzhi-cuijia: SC, Hailin: HL, and Baishan: BS) belong to S. tridactyla. This indicates the monophyly of Salamandrella and each of the two species. There was a substantial level of genetic differentiation between different species and within populations of the same species. This differentiation was significantly related to geographical distance. At last, the mismatch distribution and neutrality analyses indicated that the TL populations have undergone expansion of history. The study supplements the distributional range of Schrenck salamander. And it provides a theoretical basis for species conservation of Salamandrella species.

Buffer solution induced highly crystalline sodium-rich Prussian blue for sodium storage.

In this work, we have developed an efficient method to synthesize Prussian blue by self-decomposition of sodium ferrocyanide in acetic acid-sodium acetate buffer solution. This buffer solution-based proton pool provides a relatively low and stable concentration of protons for the slow decomposition of sodium ferrocyanide to get highly crystalline and sodium rich Prussian blue, which can be used as the cathode for high-performance sodium-ion batteries.

Quality Evaluation of Walnuts from Different Regions in China.

This study analyzed and evaluated the basic crude fat contents, crude protein contents, phenolic compounds, lipid compositions (fatty acids, phytosterols, and tocopherols), and amino acid compositions of 26 walnut samples from 11 walnut-growing provinces in China. The results indicate that the oil contents of the samples varied from 60.08% to 71.06%, and their protein contents ranged from 7.26 g/100 g to 19.50 g/100 g. The composition of fatty acids corresponded to palmitic acid at 4.61-8.27%, stearic acid at 1.90-3.55%, oleic acid at 15.50-32.28%, linoleic acid at 53.44-67.64%, and α-linolenic acid at 2.45-12.77%. The samples provided micronutrients in widely varying amounts, including tocopherol, phytosterol, and total phenolic content, which were found in the walnut oil samples in amounts ranging from 356.49 to 930.43 mg/kg, from 1248.61 to 2155.24 mg/kg, and from 15.85 to 68.51 mg/kg, respectively. A comprehensive evaluation of walnut oil quality in the samples from the 11 provinces using a principal component analysis was conducted. The findings revealed that the samples from Henan, Gansu, and Zhejiang had the highest composite scores among all provinces. Overall, Yunnan-produced walnuts had high levels of crude fat, polyunsaturated fatty acids, and total tocopherols, making them more suitable for producing high-quality oil, whereas Henan-produced walnuts, although lower in crude fat, had a higher crude protein content and composite score, thus showing the best walnut characteristics.

[Identification of heat shock protein hsp70 family genes from Rana amurensis and its expression profiles upon infection].

Heat shock proteins (HSPs) widely exist in all organisms, the structures of which are usually extraordinarily conservative. They are also well-known stress proteins that are involved in response to physical, chemical and biological stresses. HSP70 is an important member of the HSPs family. In order to study the roles of amphibians HSP70 during infection, the cDNA sequence of Rana amurensis hsp70 family genes were cloned by homologous cloning method. The sequence characteristics, three-dimensional structure and genetic relationship of Ra-hsp70s were analyzed by bioinformatics methods. The expression profiles under bacterial infection were also analyzed by real-time quantitative PCR (qRT-PCR). Expression and localization of HSP70 protein were tested by immunohistochemical techniques. The results showed that three conservative tag sequences of HSP70 family, HSPA5, HSPA8 and HSPA13, were found in HSP70. Phylogenetic tree analysis indicated four members are distributed in four different branches, and members with the same subcellular localization motif are distributed in the same branch. The relative expression levels of the mRNA of four members were all significantly upregulated (P < 0.01) upon infection, but the time for up-regulating the expression levels were diverse in different tissues. The immunohistochemical analysis showed that HSP70 was expressed to different degrees in the cytoplasm of liver, kidney, skin and stomach tissue. The four members of Ra-hsp70 family have ability to respond bacterial infection to varying degrees. Therefore, it was proposed that they are involved in biological processes against pathogen and play different biological functions. The study provides a theoretical basis for functional studies of HSP70 gene in amphibians.

[Cloning of adipor1 and adipor2 genes in Rana dybowskii and its expression pattern upon infection].

Adiponectin receptor 1 (AdipoR1) and Adiponectin receptor 2 (AdipoR2) can bind to adiponectin (AdipoQ) secreted by adipose tissue to participate in various physiological functions of the body. In order to explore the role of AdipoR1 and AdipoR2 in amphibians infected by Aeromonas hydrophila (Ah), the genes adipor1 and adipor2 of Rana dybowskii were cloned by reverse transcription-polymerase chain reaction (RT-PCR) and analyzed by bioinformatics. The tissue expression difference of adipor1 and adipor2 was analyzed by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR), and an inflammatory model of R. dybowskii infected by Ah was constructed. The histopathological changes were observed by hematoxylin-eosin staining (HE staining); the expression profiles of adipor1 and adipor2 after infection were dynamically detected by qRT-PCR and Western blotting. The results show that AdipoR1 and AdipoR2 are cell membrane proteins with seven transmembrane domains. Phylogenetic tree also shows that AdipoR1 and AdipoR2 cluster with the amphibians in the same branch. qRT-PCR and Western blotting results show that adipor1 and adipor2 were up-regulated at different levels of transcription and translation upon Ah infection, but the response time and level were different. It is speculated that AdipoR1 and AdipoR2 participate in the process of bacterial immune response, providing a basis for further exploring the biological functions of AdipoR1 and AdipoR2 in amphibians.

DNA barcoding of Chinese snakes reveals hidden diversity and conservation needs.

DNA barcoding has greatly facilitated studies of taxonomy, biodiversity, biological conservation, and ecology. Here, we establish a reliable DNA barcoding library for Chinese snakes, unveiling hidden diversity with implications for taxonomy, and provide a standardized tool for conservation management. Our comprehensive study includes 1638 cytochrome c oxidase subunit I (COI) sequences from Chinese snakes that correspond to 17 families, 65 genera, 228 named species (80.6% of named species) and 36 candidate species. A barcode gap analysis reveals gaps, where all nearest neighbour distances exceed maximum intraspecific distances, in 217 named species and all candidate species. Three species-delimitation methods (ABGD, sGMYC, and sPTP) recover 320 operational taxonomic units (OTUs), of which 192 OTUs correspond to named and candidate species. Twenty-eight other named species share OTUs, such as Azemiops feae and A. kharini, Gloydius halys, G. shedaoensis, and G. intermedius, and Bungarus multicinctus and B. candidus, representing inconsistencies most probably caused by imperfect taxonomy, recent and rapid speciation, weak taxonomic signal, introgressive hybridization, and/or inadequate phylogenetic signal. In contrast, 43 species and candidate species assign to two or more OTUs due to having large intraspecific distances. If most OTUs detected in this study reflect valid species, including the 36 candidate species, then 30% more species would exist than are currently recognized. Several OTU divergences associate with known biogeographic barriers, such as the Taiwan Strait. In addition to facilitating future studies, this reliable and relatively comprehensive reference database will play an important role in the future monitoring, conservation, and management of Chinese snakes.

[Cloning and tissue expression analysis of the LepROT gene of Rana dybowskii].

Leptin receptor overlapping transcript (LepROT) plays multiple roles in the regulation of immune systems. However, very little information is available about the anti-infectious mechanisms of amphibians LepROT. In this study, the cDNA sequence of the Rana dybowskii LepROT gene was determined by using RT-PCR and bioinformatics analysis. Then, the Aeromonas hydrophila (Ah) and lipopolysaccharides (LPS) infected models of R. dybowskii was constructed to obtain histopathological characteristics. Constitutive expression of LepROT mRNA and NF-κB signaling pathway were detected by real-time quantitative PCR. The full-length cDNA of LepROT gene was 396 bp and encoded 131 amino acids. Amino acid sequence analysis revealed LepROT shares 93.74% and 86.39% identity with homologues from other amphibians and mammals respectively, and the LepROT gene was quite conserved among different species. After infection, the relative expression levels of LepROT, NF-κB, IKKα and IKKß mRNA were all significantly upregulated (P < 0.01), but showed a diverse temporal pattern of up-regulation in different tissues. Therefore, it was proposed that the LepROT gene of R. dybowskii might activate the NF-κB signaling pathway to exert anti-infectious effects, thus providing evidence for further extending the biological function of LepROT.

Microbial composition of carapace, feces, and water column in captive juvenile green sea turtles with carapacial ulcers.

Introduction: Green sea turtles are endangered marine reptiles. Carapacial ulcers will develop on juvenile green sea turtles during artificial rescue, seriously affecting their health and potentially leading to death. Methods: To determine the pathogens causing ulcerative carapacial disease, we performed 16S and ITS high-throughput sequencing, and microbial diversity analysis on samples from carapacial ulcers, healthy carapaces, feces, and seawater of juvenile green sea turtles. Results: Our analysis showed that changes in microbial diversity of green sea turtle feces and seawater were not significantly associated with ulcerative carapacial disease. Discussion: Psychrobacter sp. is the dominant species in the carapacial ulcers of green sea turtles. The bacterium is present in both healthy turtles and seawater where carapacial ulcers did not occur and decreasing seawater temperatures are likely responsible for the infection of juvenile green turtles with Psychrobacter sp. This is the first study on carapacial ulcers in captive juvenile green sea turtles. Our research provides theoretical guidance for the prevention and control of carapacial ulcers in captive juvenile green sea turtles.

[Expression of MHCâ genes in different tissues of Rana dybowskii under the stress of Aeromonas hydrophila].

The aim of this study was to investigate the expression of MHCⅠ gene in different tissues of Rana dybowskii under the stress of Aeromonas hydrophila (Ah), and to provide evidence for revealing the anti-infective immune response mechanism of amphibians. The experimental animal model of Aeromonas hydrophila infection was first constructed, and the pathological changes were observed by HE staining. The MHCⅠ gene α1+α2 peptide binding region of Rana dybowskii was cloned by RT-PCR and analyzed by bioinformatics. Real-time PCR was used to detect the transcription level of MHCⅠ in different tissues under Ah stress. After Ah infection, the skin, liver and muscle tissues showed signs of cell structure disappearance and texture disorder. The MHCⅠ gene α1+α2 peptide binding region fragment was 494 bp, encoding 164 amino acids, and homology with amphibians. Above 77%, the homology with mammals was as low as 14.96%, indicating that the α1+α2 region of MHC gene was less conserved among different species. The results of real-time PCR show that the liver, spleen and kidney of the experimental group were under Ah stress. The transcript levels of MHCⅠ gene in skin and muscle tissues were higher than those in the control group at 72 h, but the time to peak of each tissue was different (P<0.01), indicating that the response time of MHCⅠ gene in different tissues was different under Ah stress. This study provides a reference for further exploring the immune function of MHC molecules in anti-infection.

Genetic diversity of major histocompatibility complex class I genes in Zootoca vivipara.

The Major Histocompatibility Complex (MHC), as a family of highly polymorphic genes associated with immunity in the genome of the vertebrate, has become an important indicator for assessing the evolutionary potential of wildlife. In order to better protect Zootoca vivipara in the Greater Khingan Range and Lesser Khingan Range, to understand the genetic structure of Z. vivipara, and to explore the mechanism and phylogenetic relationship of the gene polymorphisms, the MHC molecular marker method was used to analyze Z. vivipara population. Forty-seven alleles were obtained from four populations. The four populations were highly polymorphic, rich in genetic information, and had significant genetic diversity. There were certain inbreeding phenomena. There was a high degree of genetic differentiation among populations, which was caused by genetic drift and natural selection. The sequence undergoes genetic duplication and recombination. The existence of trans-species polymorphism was found in the constructed phylogenetic tree. The present study provides a theoretical basis for species protection of Z. vivipara.

Impact of endometriosis on risk of ovarian, endometrial and cervical cancers: a meta-analysis.

PURPOSE: The risks of gynecologic cancer have not been well established in women with endometriosis. The objective of the present study was to investigate the influence of endometriosis on the risk for three gynecologic cancer (ovarian cancer, endometrial cancer and cervical cancer). METHODS: We gathered updated evidence about the risk relationship between endometriosis and gynecologic cancers by conducting a comprehensive search of several medical literature electronic databases, including PubMed, Embase and the Cochrane Library. The design and quality of all studies were evaluated using the Newcastle-Ottawa Scale (NOS), and a random-effects model was used to calculate pooled risk ratio (RR). RESULTS: Of the 8538 articles our search produced, we selected 25 qualified studies, including 16 cohort studies and 9 case-control studies. Patients with endometriosis had both an increased risk of ovarian cancer [RR 1.964; 95% CI (1.685, 2.290)]. The risk of endometrial cancer (EC) is not necessarily higher in patients with endometriosis [RR 1.176, 95% CI (0.878, 1.575)]. Endometriosis was not associated with an increased risk for cervical cancer (CC) [RR 0.670, 95% CI (0.537, 0.838)]. CONCLUSIONS: Patients with endometriosis need to be closely observed and rechecked regularly to prevent malignant changes.

Comparative effectiveness of different chemotherapy regimens of advanced-stage Hodgkin lymphoma in adults: a network meta-analysis.

BACKGROUND: Combined chemotherapy is the cornerstone treatment for patients with advanced Hodgkin lymphoma (HL). The objective of our study was to perform a network meta-analysis of the efficacy of different chemotherapy regimens in adults with advanced-stage HL. MATERIALS AND METHODS: We searched for relevant randomized controlled trials (RCTs) in titles/abstracts in PubMed, Embase, and the Cochrane Library. The search was last updated on April 3, 2018. RCTs that assessed the effectiveness of one of the following treatments were included: doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD); four cycles of increased dose of bleomycin, etoposide, doxorubicin, cyclophosphamide, vincristine, procarbazine, and prednisone (BEACOPPescalated) followed by two or four cycles of standard dose of BEACOPP (4× BEACOPPescalated + 2 or 4× BEACOPPbaseline); brentuximab vedotin plus doxorubicin, vinblastine, and dacarbazine (A+AVD); doxorubicin, vinblastine, mechlorethamine, vincristine, bleomycin, etoposide, and prednisone combined with radiation therapy (Stanford V); mechlorethamine (cyclophosphamide), vincristine, procarbazine, and prednisone (M[C] OPP); sequential or alternating chemotherapy regimens with ABVD as the footstone (eg, COPP/ABVD or mechlorethamine, vincristine, procarbazine, and prednisone [MOPP]/ABVD); eight cycles of BEACOPPescalated; hybrid MOPP/ABV; and M[C]EC (M[C]OPP with epidoxorubicin, bleomycin, vinblastine [EBV], and lomustine, doxorubicin, and vindesine [CAD]). RESULTS: Overall, we screened 3,564 citations and deemed 18 reports of 16 trials eligible and included them in our network meta-analysis. A total of 11,928 participants were randomly assigned to one of the 12 combinations of chemotherapy regimens, of which 11,476 participants were analyzed. For the overall survival (OS), no differences were observed within any interventions when the ABVD regimen was used as the reference treatment. Similarly, relative to A+AVD, 8× BEACOPPescalated and 6× BEACOPPescalated also showed no differences (HR =1.07, 95% credible interval (CrI): 0.58-1.95; HR =0.62, 95% CrI: 0.16-1.83; and HR =0.71, 95% CrI: 0.30-1.72, respectively). In terms of complete remission (CR), enough evidence exists to support a maximum clinical treatment effect for 6× BEACOPPescalated (OR =1.88, 95% CrI: 1.20-2.96; and OR =3.43, 95% CrI: 1.87-6.24). CONCLUSION: When compared across the 12 combined chemotherapy regimens, six cycles of BEACOPPescalated may be the optimal treatment for patients with advanced-stage HL.

Comparative efficacy of different targeted therapies plus fulvestrant for advanced breast cancer following progression on prior endocrine therapy: a network meta-analysis.

BACKGROUND: We performed a network meta-analysis of randomized controlled trials (RCTs) to indirectly compare the efficacy of different targeted agents with fulvestrant for patients with hormone-receptor-positive (HR+) and human epidermal growth factor receptor type 2-negative (HER2-) advanced breast cancer (ABC) following progression on prior endocrine therapy. METHODS: The titles/abstracts were searched from the PubMed, EMBASE, and the Cochrane Library databases for RCTs to evaluate the efficacy of palbociclib plus fulvestrant vs alternative targeted therapies plus fulvestrant for postmenopausal HR+/HER2- ABC following progression on prior endocrine therapy. In addition, the primary measured outcome was progression-free survival (PFS) and objective response rate. The surface under the cumulative ranking (SUCRA) value of each treatment was calculated to achieve the best ranking for each treatment. RESULTS: A total of 11 studies, including 4,178 patients in the network meta-analysis, were included and analyzed. In terms of the pooled hazard ratios (HRs) for PFS, palbociclib plus fulvestrant was superior to other target agents plus fulvestrant (HR=0.62, 95% credible interval [CrI]: 0.40-0.96; HR=0.62, 95% CrI: 0.47-0.96; for pictilisib plus fulvestrant and buparlisib plus fulvestrant, respectively). Ribociclib plus fulvestrant has no difference in abemaciclib plus fulvestrant and palbociclib plus fulvestrant (HR =1.02, 95% CrI =0.72-1.45; HR =1.22, 95% CrI =0.84-1.78). In terms of objective response rate, compared with placebo plus fulvestrant, abemaciclib plus fulvestrant, dovitinib plus fulvestrant, buparlisib plus fulvestrant, and palbociclib plus fulvestrant had a significant difference (odds ratio [OR] =2.84, 95% CrI =1.91- 4.31; OR =3.62, 95% CrI =1.21-12.48; OR =1.80, 95% CrI =1.25-2.60; and OR =2.52, 95% CrI =1.43- 4.72, respectively). CONCLUSION: According to the present study, palbociclib plus fulvestrant may be the optimal treatment for HR+/HER2- postmenopausal women with ABC after disease progression following endocrine therapy.

The association between statin use and endometrial cancer survival outcome: A meta-analysis.

BACKGROUND: Previous studies on the association between statin use and survival outcomes in gynecologic cancers have presented conflicting results. No independent studies to elucidate the association between statin use and survival outcomes of endometrial cancer (EC) have been conducted. METHODS: To gather updated evidence, we carried out an extensive literature search on Medline (PubMed and OvidSP), Embase, Cochrane Library, China National Knowledge Infrastructure (CNKI), wanfang data, and Vip network to identify all potential studies on the effect of statins on the prognosis of endometrial carcinoma. The design and quality of all studies were evaluated, and a fixed-effects model was used to calculate pooled hazard ratios (HRs) for overall survival (OS) and disease-specific survival (DSS). RESULTS: Of the 219 articles screened, 9 articles were eligible, including 8 articles and 1 abstract. A total of 5923 patients with endometrial cancer who used statins were identified. Statin use was related to increased overall survival (HR, 0.80; 95% confidence interval [CI], 0.66-0.95, without significant heterogeneity, I = 52%, P = .080). Statin users also had increased disease-specific survival (HR, 0.69; 95% CI, 0.61-0.79, I = 0.0%). CONCLUSION: Statins are beneficial to the survival outcome of patients with endometrial cancer. The selection of statins as a 1st-line agent seems justified for endometrial carcinoma.

Effect of first-line endocrine therapy in patients with hormone-sensitive advanced breast cancer: a network meta-analysis.

BACKGROUND: Endocrine therapy is the cornerstone treatment for patients with hormone receptor-positive advanced breast cancer. We aimed to assess the effectiveness of various first-line endocrine monotherapies or combinations to determine the optimal sequence in a network meta-analysis. MATERIALS AND METHODS: We searched PubMed, EMBASE, and the Cochrane Library for randomized controlled trials (RCTs) from inception up to November 21, 2017. We included only RCTs that assessed the effectiveness of the following treatments as a monotherapy or in combination as the first-line treatment: tamoxifen, anastrozole, letrozole, exemestane, fulvestrant, palbociclib, and ribociclib. The results were presented with pooled odds ratio or hazard ratio (HR), and 95% credible interval (CrI). The primary outcomes were objective response rate (ORR) and progression-free survival/time to progression. RESULTS: A total of 16 eligible articles (14 RCTs) involving 6,602 patients treated with 10 different first-line endocrine therapies were assessed in our network meta-analysis. Palbociclib plus letrozole was superior to anastrozole, letrozole, exemestane, fulvestrant 500 mg, and anastrozole plus fulvestrant (loading dose) (HR=0.44, 95% CrI: 0.33-0.58; HR=0.56, 95% CrI: 0.45-0.68; HR=0.45, 95% CrI: 0.32-0.61; HR=0.58, 95% CrI: 0.42-0.81; HR=0.50, 95% CrI: 0.37-0.68; respectively). However, there is no significant advantage compared with ribociclib plus letrozole (HR=1.00, 95% CrI: 0.72-1.39). In terms of ORR, ribociclib plus letrozole is more effective than palbociclib plus letrozole (odds ratio=1.30, 95% CrI: 0.83-2.02). CONCLUSION: Palbociclib plus letrozole and ribociclib plus letrozole might be the optimal first-line endocrine therapeutic choices for hormone receptor-positive/human epidermal growth factor receptor 2-negative advanced breast cancer due to a longer progression-free survival/time to progression and a more efficacious ORR.

Clinicopathological characteristics and prognostic value of the cancer stem cell marker ALDH1 in ovarian cancer: a meta-analysis.

BACKGROUND: The clinicopathological and prognostic values of the cancer stem cell marker aldehyde dehydrogenase 1 (ALDH1) in ovarian cancer (OC) remain unknown. The aim of our meta-analysis was to evaluate ALDH1's association with clinicopathological characteristics and its prognostic significance in patients with OC. MATERIALS AND METHODS: PubMed, Embase, and China Biology Medicine were systematically searched for eligible studies (up to October 2017). Pooled odds ratios (ORs) or hazard ratios (HRs) with 95% CIs were used to evaluate the association of ALDH1 expression with clinicopathological features and survival outcomes. RESULTS: A total of 17 papers (18 studies) that included 2,531 patients with OC were analyzed. The results showed a significant association between increasing ALDH1 expression and International Federation of Gynecology and Obstetrics stage (OR 2.02, 95% CI 1.16-3.52), lymph node metastasis (OR 1.91, 95% CI 1.01-3.61), and distant metastasis (OR 5.43, 95% CI 1.44-20.42) in OC. However, no significant correlation was found between increasing ALDH1 expression and age (OR 0.90, 95% CI 0.25-3.28), tumor size (OR 1.13, 95% CI 0.75-1.71), tumor location (OR 0.69, 95% CI 0.22-2.13), ascite status (OR 0.74, 95% CI 0.49-1.11), resistance status (OR 0.70, 95% CI 0.14-3.51), or clinicopathological type (OR 1.14, 95% CI 0.69-1.86). Moreover, a high ALDH1 expression was significantly associated with overall survival (HR 1.56, 95% CI 1.21-2.02) but not with disease-free survival (HR 1.38, 95% CI 0.99-1.93). CONCLUSION: The meta-analysis indicates that increasing ALDH1 predicts poor prognosis and clinicopathological characteristics in OC. Future studies are needed to explore tailored treatments that directly target ALDH1 for the improvement of survival in OC.

Effects of HIV on metabolic and biological pathways of CD4+ T lymphocytes.

The effects of human immunodeficiency virus (HIV) on the metabolic and biological pathways of cluster of differentiation (CD)4+ T lymphocytes were investigated. A total of 150 patients with acquired immune deficiency syndrome (AIDS) and 50 healthy individuals who were admitted to hospital for physical examination during the period of June 2016 to January 2017, were selected as subjects in the present study. According to the virus load, 150 AIDS patients were divided into three groups: i) Viral load >106 copies/ml (group A, n=39), ii) 104 copies/ml < viral load <105 copies/ml (group B, n=76), and iii) viral load <104 copies/ml (group C, n=35). The relationship between viral loads in the three groups and CD4+ T lymphocyte counts was assessed. Active lymphocytes were isolated from T lymphocytes in the subjects, and the ratio of Th1 to Th2 was measured by flow cytometry. Effects of HIV on human T-lymphocyte differentiation were observed. Differences in T-lymphocyte metabolites were detected by proton nuclear magnetic resonance and their biological pathways analyzed. The results showed that CD4+ T-cell counts were decreased with the increase of the viral loads of patients. The viral loads of AIDS patients differentiated T lymphocytes. In other words, high viral loads accelerated the differentiation of T lymphocytes into Th1 cells. In the high HIV viral load group, the levels of glycerol phosphodiesterase, 7-dehydrocholesterol, p-hydroxyphenylacetic acid, cholesterol and deoxyuridine were increased, but the levels of 3-methoxytyramine, cytidine deaminase, deoxycorticosterone and 3-hydroxybutyric acid were decreased. The viral loads of AIDS patients are associated with CD4+ T-cell counts and the ratio of CD4+ T to CD8+ T cells. At the same time, HIV viral loads can affect the lipid biosynthesis of T-lymphocyte membranes, thus affecting the differentiation and proliferation of T lymphocytes and finally intervening its mediated immune responses.

Role of miR-124a in T cell activation and immunity in AIDS patients.

The role of microRNA-124a (miR-124a) in the regulation of T cell activation and immunity in patients with AIDS, was studied to provide new insights for the study, diagnosis, alleviation and treatment of AIDS. RT-qPCR technique was used to quantitatively analyze the expression of miR-124a in peripheral blood CD4+ T cells. Dual-luciferase reporter assay system was established to report possible regulatory relations between miR-124a and its potential target gene SIRT1. RT-qPCR and western blot analysis were used to detect the expression level of mRNA and protein of the target genes in T cells. Normal CD4+ T cells from controls were transfected with miR-124a mimics and its negative control, and miR-124a inhibitor and its negative control were transfected into CD4+ T cells from patients with AIDS by T lymphocyte transfection kit to detect the relative expression level of SIRT1 mRNA and protein. The levels of interferon (IFN)-γ, interleukin (IL)-10, transforming growth factor (TGF)-ß, IL-2, IL-4 and IL-6 secreted by T helper cells were detected by enzyme-linked immunosorbent assay (ELISA). miR-124a was upregulated in CD4+ T cells of patients with AIDS. The results of firefly luciferase activity detection showed that miR-124a can directly interact with target gene SIRT1 and negatively regulate its expression. miR-124a mimics/inhibitor transfection experiments showed that overexpression of miR-124a in normal CD4+ T cells significantly reduced SIRT1 expression compared with control group, and the expression of miR-124a was positively correlated with IL-10 and TGF-ß expression and negatively correlated with IFN-γ expression, but showed no correlation with other cytokines. In AIDS patients, the inhibition of expression of miR-124a in CD4+ T cells significantly increased the expression of SIRT, at the same time, the expression levels of IL-10 and TGF-ß were significantly decreased, while the expression level of IFN-γ was significantly increased and no significant difference was found in the expression of other cytokines. The expression of miR-124a in CD4+ T cells of AIDS patients was upregulated and the Th2 type CD4+ T cells are activated by SIRT1 expression inhibition, which in turn enhance the immunity of HIV-infected cells. Our study provides a new molecular target for the diagnosis, alleviation and treatment of AIDS.

Expression of Foxp3 in renal tissue of patients with HBV-associated glomerulonephritis and their clinical and pathological characteristics.

Our study retrospectively investigated the expression of forkhead/winged helix transcription factor (Foxp3) in renal tissue and clinical features of patients with hepatitis B virus (HBV)-associated glomerulonephritis (HBV-GN). A total of 58 patients with HBV-GN were assigned to group A; 45 serum and renal tissue HBsAg-negative patients with nephritis were group B; 24 serum HBsAg-positive and renal tissue HBsAg-negative patients with slightly increased serum creatinine without nephritis were group C. Clinical manifestations, laboratory indices and renal biopsies were recorded. Expression of Foxp3, CD4 and CD25 in renal tissue was detected by immunohistochemistry. In group A, 74.1% were serum HBeAg-negative, with serum complement C3 level of 0.99±0.27 g/l, and deposition rates of renal complement C3 and C1q in renal tissue of 34.9 and 16.3% respectively; 25.9% were serum HBeAg-positive, with serum complement C3 level of 0.19±0.17 g/l, and deposition rates of renal complement C3 and C1q in renal tissue of 80 and 46.7%, respectively. A significant difference was found in C3 and C1q between HBeAg-negative and HBeAg-positive group (P<0.05). Increased urinary protein and decreased serum albumin were found in patients in group A with moderate levels of HBV DNA compared with patients with low levels of HBV DNA in the same group over 24 h (P<0.05). The numbers of Foxp3+ lymphocytes, CD4+ T cells and CD25+ T cells in the tubulointerstitium of patients in groups A and B were 3.41±1.16 vs. 3.52±1.27, 2.78±0.15 vs. 3.12±0.17 and 2.90±0.20 vs. 3.09±0.18, respectively. The clinical manifestation of HBV-GN is nephrotic syndrome, and HBV DNA is correlated with urinary protein and serum albumin levels. Activation of C3 and C1q may be related to the pathogenesis of HBV-GN in serum HBeAg-positive patients. Downregulation of Foxp3 expression in regulatory T cells is related to the development and progression of HBV-GN.

Sequencing and analysis of the complete mitochondrial genome of Hyla ussuriensis (Anura: Hylidae).

In this study, the complete mitogenome sequence of Hyla ussuriensis (Anura: Hylidae) is first determined using long PCR. It is a circular molecule of 18 023 bp in length (GenBank accession no. KT964710). Similar to the typical mtDNA of amphibians, the complete mtDNA sequence of Hyla ussuriensis contained two rRNA genes (12S rRNA and 16S rRNA), 22 tRNA genes, 13 protein-coding genes (PCGs), and a control region (D-loop). The nucleotide composition was 29.9% A, 25.4% C, 14.5% G, and 30.2% T. Mitochondrial genomes analyses based on NJ method yield phylogenetic trees, indicating 13 reported Anura frogs belonging to five families (Hylidae, Bufonidae, Microhylidae, Ranidae, and Rhacophoridae). These molecular data presented here provide a useful tool for systematic analyses of genus Hyla and family Hylidae.