Loop-mediated isothermal amplification linked a nanoparticles-based biosensor for detecting Epstein-Barr virus.

Epstein-Barr virus (EBV) is a ubiquitous gamma herpesvirus that maintains a lifelong latent association with B lymphocytes. Here, a rapid and reliable diagnosis platform for detecting EBV infection, employing loop-mediated isothermal amplification (LAMP) combined with a gold nanoparticles-based lateral flow biosensors (AuNPs-LFB) (termed LAMP Amplification Mediated AuNPs-LFB Detection, LAMAD), was developed in the current study. A set of specific LAMP primers targeting the Epstein-Barr nuclear antigen (EBNA) leader protein (EBNA-LP) gene was designed and synthesized. Subsequently, these templates extracted from various pathogens and whole blood samples were used to optimize and evaluate the EBV-LAMAD assay. As a result, the limit of detection (LoD) of the EBV-LAMAD assay was 45 copies/reaction. The EBV-LAMAD assay can detect all representative EBV pathogens used in the study, and of note, no cross-reactions were observed with other non-EBV organisms. Moreover, the whole workflow of the EBV-LAMAD assay can be completed within 70 min, including rapid EBV template preparation, EBV-LAMP amplification, and AuNPs-LFB-mediated detection. Taken together, the EBV-LAMAD assay targeting the EBNA-LP gene is a rapid, simplified, sensitive, reliable, and easy-to-use detection protocol that can be used as a competitive potential diagnostic/screening tool for EBV infection in clinical settings, especially in basic laboratories in resource-limited regions. KEY POINTS: • A novel, simplified, and easy-to-use AuNPs-LFB biosensor was designed and prepared. • LAMP combined with an AuNPs-LFB targeting the novel EBNA-LP gene was established. • EBV-LAMAD is a rapid, sensitive, and reliable detection protocol for EBV infection.

Trends of HIV/Syphilis/HSV-2 seropositive rate and factors associated with HSV-2 infection in men who have sex with men in Shenzhen, China: A retrospective study.

OBJECTIVES: To analyze the trends of HIV/syphilis/HSV-2 seropositive rate and explore the related factors with HSV-2 infection to provide the basis for adjusting STD intervention strategies and formulating prevention and control measures among MSM in Shenzhen. METHODS: Time-location sampling was conducted among MSM in Shenzhen in 2012, 2014, 2016, and 2018. Data on demographics, sexual behaviors and the laboratory test results of HIV, syphilis, HSV-2 were collected. The χ2 trend test was used to analyze the trends of HIV/syphilis/HSV-2 seropositive rate. The binary logistic regression model was used to explore the factors associated with HSV-2 infection. RESULTS: The seropositive rate of HIV fell significantly from 15.9% in 2012 to 8.7% in 2018 (Ptrend = 0.003), syphilis seropositive rate was significantly decreased from 20.4% in 2012 to 14.8% in 2018 (Ptrend = 0.025), HSV-2 seropositive rate had no significant change (16.7% in 2012 to 14.0% in 2018; Ptrend = 0.617). In principal component logistic regression analysis showed that FAC1_1 (X1 = Ever had sex with female, X2 = Gender of first sexual partner, X3 = Marital status, X4 = Age group), FAC2_1 (X5 = Education, X6 = Monthly income (RMB), X7 = Frequency of condom use in anal sex with men in the past 6 months), and FAC4_1 (X9 = History of STDs) were significantly associated with HSV-2 infection. CONCLUSIONS: The seropositive rates of HIV and syphilis have dropped significantly but are still high. HSV-2 seropositive rate had no significant change and maintained a high level. It is necessary to continue strengthening HIV and syphilis interventions among MSM in Shenzhen. HSV-2 detection and intervention are urgently required for MSM, which might be another effective biological strategy further to control the HIV epidemic among MSM in Shenzhen.

[Effects of land use change on soil organic carbon and its components in karst rocky desertification of southwest China]. / è¥¿å—å–€æ–¯ç‰¹çŸ³æ¼ åŒ–çŽ¯å¢ƒä¸‹åœŸåœ°åˆ©ç”¨å˜åŒ–å¯¹åœŸå£¤æœ‰æœºç¢³åŠå ¶ç»„åˆ†çš„å½±å“.

Soil organic carbon (SOC) is the dynamic medium of carbon transfer and the main pathway of carbon transfer in the karst ecosystem. SOC and its components are the important parts in soil carbon cycling of karst ecosystem. However, few studies have focused on SOC and its components in the karst ecosystem. We analyzed the effects of land use change on the SOC content, SOC reserve (SOCS), water-soluble organic carbon (WSOC), easily oxidizable organic carbon (EOC), particu-late organic carbon (POC), and light fraction organic carbon (LFOC), and heavy fraction organic carbon (HFOC) and their distribution ratio, with six different land-use patterns [Zanthoxylum bungeanum forest (HJ), Hylocereus undulates forest (HL), mixed forest of Z. bungeanum and H. undulates (HHL), Sabina chinensis forest (YB), mixed forest of S. chinensis and Ligustrum luci-dum (YBN), and slope cropland (PD)] in Huajiang Canyon of Guanling County, Guizhou Pro-vince. Results showed that SOC and SOCS in YB, YBN and HJ were significantly higher than those in HL, HHL and PD. In the 0-20 cm soil layer, the concentrations of SOCS followed the order of HJ＞YB＞YBN＞PD＞HHL＞HL. Contents of WSOC, EOC, POC, LFOC and HFOC in YB, YBN, and HJ were all higher than those in the other three patterns. Significant positive correlations existed between SOC and each of its components (WSOC, EOC, POC, LFOC and HFOC), also between any two of those components. Z. bungeanum could be used as a priority economic species for the ecological rehabilitation of karst rocky desertification and mountain agriculture development in Southwest China. WSOC, EOC, POC, LFOC and HFOC could be used as indicators of soil organic carbon pool.

RNA Sequencing Reveals the Suitability of Cardiac Death Livers for Transplantation.

BACKGROUND: Organ transplantation is considered the best treatment for end-stage organ failure. However, the lack of available organs for transplantation and the increasing number of patients waiting for transplants are primary issues facing the transplant community. Thus, developing strategies to increase the number of donors, especially for liver transplantation, has become a priority. The use of organs acquired from donors who suffered cardiac related deaths has increased the pool of potential liver donors. However, donation after cardiac death (DCD) livers increases the risk of primary graft dysfunction. METHODS: In the current study, we conducted transcriptome sequencing using livers from a DCD rat to assess the short-term feasibility and functional efficacy of DCD livers. RNA sequencing (RNAseq) data showed that the liver transcriptome varied greatly in rat livers subjected to 15 minutes of cardiac arrest. RESULTS: The livers used in the current study had a significant loss of normal function before transplantation. Functional and network analyses consistently indicated that transcription and translation processes were inhibited after approximately 15 minutes of cardiac arrest. Moreover, the transcriptomic sequencing data provides significant insight for identifying functional genes and testing additional biological questions in DCD liver transplantation in future studies.

Cross-cultural adaptation of the Kidney Transplant Questionnaire for Chinese adult kidney allograft recipients.

OBJECTIVE: To provide a disease-specific instrument for evaluating the health-related quality life of Chinese kidney allograft recipients. METHODS: Cross-cultural adaptation of the Kidney Transplant Questionnaire (KTQ) was performed by forward translation of the original English version into Chinese, followed by back translation and evaluation of the Chinese version by health care professionals, language professionals, and the translators. RESULTS: A total of 297 patients (110 women and 187 men; mean age, 43.91 ± 11.38 y; average time since transplant, 40.36 ± 32.86 mo) completed the Chinese versions of the KTQ and 36-Item Short Form Health Survey, and the results were used to evaluate the validity and reliability of the Chinese KTQ. The Cronbach α values for all KTQ dimensions were satisfactory (physical symptoms, α = 0.876; fatigue, α = 0.896; uncertainty/fear, α = 0.686; appearance, α = 0.701; and emotions, α = 0.886) and similar to values reported for the English and Spanish versions. The correlation coefficients among the dimensions of the Chinese KTQ ranged from 0.26 to 0.69, and those between the KTQ and 36-Item Short Form Health Survey physical component summary and mental component summary subscales were low and moderate to high, respectively, except for the appearance dimension. A good fit of the data in the confirmatory factor analysis indicated that the individual items of the translated instrument indeed evaluated the intended concepts. CONCLUSION: The Chinese version of the KTQ was found to be similarly valid and reliable compared with the original version and, thus, can be used to evaluate health-related quality of life among Chinese adult kidney allograft recipients.

Effects of isovitexin on transient outward potassium current of ventricular myocytes in rats / 中国中药杂志

To investigate the effects of isovitexin Ⅳ on transient outward potassium current in rat ventricular myocytes. In this study, MTT assay was used to investigate the safe range of isovitexin. The results showed that the IC₅₀ of the drug was in the range of 10-30 μmol•L⁻¹, and the drug concentration of 1-3 μmol•L⁻¹ for the patch clamp test was within the safe range. In addition, the single ventricular myocytes were obtained by single-enzymatic hydrolysis through aortic retrograde perfusion. The transient outward potassium current (Ito) of rat ventricular myocytes was guided and measured by whole-cell patch-clamp technique and the changes of current characteristics were recorded after isovite was applied. When the concentration of IV was less than 0.1 μmol•L⁻¹, there was no significant effect on Ito. However, with the increase in the concentration of IV (≥0.3 μmol•L⁻¹), the peak of Ito was decreased gradually, from (32.32±2.9) pA/pF to （25.83±4.3） pA/pF, 1 μmol•L⁻¹ IV and (19.51±3.5) pA/pF, 3 μmol•L⁻¹ IV respectively, with an inhibition effect in a concentration-dependent manner. In the range of 1-3 μmol•L⁻¹, IV down-regulated the I-V curve of Ito significantly. The activation curve showed that IV can enable the maximum half activation potential (V1/2) to move to the positive direction, and the V1/2 was increased from (19.59±1.6) mV to (22.81±1.7) mV and (28.86±1.4) mV at concentration of 1, 3 μmol•L⁻¹, meanwhile the activation curve moved to the right. However, the maximum half inactivating potential (V1/2) of the steady-state inactivation curve of Ito was significantly decreased from (－51.43±0.99) mV to (－61.81±1.3) mV with concentration of 1 μmol•L⁻¹ and (－71.50±1.4) mV with concentration of 3 μmol•L⁻¹. The inactivation time constant of recovery from inactivation (τ) was up-regulated significantly from (94.89±0.73) ms to (118.5±1.5) ms and (162.4±1.4) ms at concentration of 1, 3 μmol•L⁻¹ respectively. Meanwhile IV could enable the inactivation recovery curve to move to the right, which suggested that it can prolong the recovery time from inactivation of the transient outward potassium channel. In conclusion, isovitexin had a high inhibitory effect on Ito in rat ventricular myocytes.

A case-control study on reduced HDL2b in patients with polycystic ovarian syndrome.

High-density lipoprotein (HDL) is an important factor associated with the increasing risk of future ischaemic heart disease. In this study, we analyzed serum HDL2b level in the patients with polycystic ovary syndrome (PCOS). Total of 60 female patients with PCOS was enrolled for assessment and another 60 non-PCOS females with matched age and weight were selected as control. A highly sensitive microfluidic chip was employed to analyze the serum HDL subfractions. Serum HDL2b and HDL2b/HDL ratio were decreased in PCOS group than those in the control group (p < 0.05). Pearson correlation analysis revealed that serum HDL2b level was negatively correlated with body mass index, waist circumference, waist-to-hip ratio, INS0, HOMA-IR, T, estradiol, triglyceride (TG) and low-density lipoprotein (LDL)-C; and the ratio of HDL2b/HDL was negatively correlated with T, TG and LDL-C. Stepwise multiple regression analyses showed a reverse correlation for HDL2b and its ratio to HDL with hyperandrogenism. The results suggested that a reduction of serum HDL2b and its ratio to total serum HDL in PCOS patients by using the microfluidic chip method assessment. Hyperandrogenism was the main factor to affect HDL2b and its ratio to total HDL in PCOS patients, and it might increase the incidence of atherosclerosis as well as the risk of coronary heart disease.

Pilot feasibility research of Chinese version of kidney transplant questionnaire in recipients of living donor kidney transplantation.

OBJECTIVE: To explore the feasibility of the Chinese version of Kidney Transplant Questionnaire (KTQ) by evaluating the health-related quality of life (HRQoL) in Chinese recipients of living donor kidney transplantation. METHODS: The English version of KTQ was translated into Chinese and underwent cultural adaptation to obtain the Chinese version of KTQ. HRQoL of 136 Chinese recipients of living donor kidney transplantation that met the inclusion criteria were evaluated to assess the validity and reliability of the questionnaire. RESULTS: One hundred and thirty-six recipients (98 males and 38 females) of living donor kidney transplantation were included. The mean age of the recipients was 43.91 years. For each dimension of the questionnaire, the Cronbach's alpha coefficient was 0.7-0.9, test-retest reliability coefficient ≥0.7, goodness of fit index (GFI) >0.9, and comparative fitness index (CFI) >0.9. CONCLUSION: The validity and reliability of the Chinese version of KTQ is similar to the English version, suggesting that the Chinese version of KTQ could be applied as a disease-specific questionnaire to evaluate the HRQoL of the recipients of living donor kidney transplantation in China.

α-Fetoprotein is a potential survival predictor in hepatocellular carcinoma patients with hepatitis B selected for liver transplantation.

BACKGROUND: Risk factors can affect candidacy and prognosis following orthotopic liver transplantation (OLT) with antiviral prophylaxis for the treatment of hepatocellular carcinoma (HCC) associated with hepatitis B virus (HBV) and cirrhosis. OBJECTIVE: The objective of this study was to investigate the risk factors affecting OLT outcomes in patients with HCC/HBV-induced cirrhosis selected by two contemporary candidacy strategies. PATIENTS AND METHODS: From July 2002 to December 2006, 203 patients with HCC/HBV-cirrhosis undergoing OLT with antiviral prophylaxis were evaluated retrospectively. Patients with uncomplicated HCC fulfilling Milan (conservative candidacy group) or Up-to-Seven but not Milan (inclusive candidacy group) criteria were included. Patients received postoperative immunosuppressive therapy. Tumor-free survival and overall survival (OS) were assessed. Univariate analyses between OS and clinical/demographic factors were carried out, including α-fetoprotein (AFP), aspartate aminotransferase, alanine aminotransferase, tumor size, tumor nodule number, vascular invasion, lymph node metastasis, and degree of differentiation. OS was compared between the three groups on the basis of AFP level (≤20, 20-200, and >200 ng/ml). RESULTS: Conservative candidacy group OS and tumor-free survival were better than the inclusive candidacy group. Low AST, high tumor differentiation, and low AFP were significantly associated with improved OS in the inclusive candidacy group (P<0.05). Low tumor nodule number and AFP levels were significantly associated with improved OS in the conservative candidacy group (P<0.05). AFP of more than 200 ng/ml indicated poorer outcomes in all groups. In multivariate analysis, AFP was an independent predictor of OS. CONCLUSION: Up-to-Seven criteria may be more appropriately stratified by AFP, AST, and tumor differentiation, and AFP is a potential independent survival predictor in HBV-associated HCC patients selected for OLT.

[Expression of PPARγ mRNA in granulosa cells and its correlation with clinical characteristics of polycystic ovary syndrome].

OBJECTIVE: To study the expression of PPARγ mRNA in granulosa cells of patients with polycystic ovary syndrome (PCOS) and the impact of testosterone, insulin and PPARγ agonist rosiglitazone on granulosa cells (GCs). METHODS: The expression of PPARγ mRNA in GCs of patients with PCOS and normal controls were analyzed by Real-time PCR. We assessed the level of PPARγ mRNA in GCs from normal controls after treatment with testosterone, insulin, and rosiglitazone. RESULTS: The expression of PPARγ mRNA was lower in the GCs of PCOS than that of the controls (P<0.05). When testosterone concentration was 1 nmol/L, the expression of PPARγ mRNA was lower in the GCs as compared with the blank control (P<0.05). When testosterone concentration was 10 nmol/L, PPARγ mRNA increased in the GCs as compared with the blank control, which was of no significance (P>0.05). When insulin concentration was 10 nmol/L, the expression of PPARγ mRNA was higher in the GCs as compared with the blank control (P<0.05). When insulin concentration was 100 nmol/L, the expression of PPARγ mRNA increased, but the difference was not statistically significant (P>0.05). When rosiglitazone concentration was 1 nmol/L, the expression of PPARγ mRNA in ovarian GCs significantly increased, as compared with the blank control (P<0.05). When rosiglitazone concentration was at 10 nmol/L, the PPARγ mRNA expression significantly increased, as compared with the concentration at 1 nmol/L (P<0.05). CONCLUSION: PPARγ mRNA expression is down-regulated by testosterone, and up-regulated by insulin and rosiglitazone with different dosages. Decreased PPARγ mRNA in the GCs of PCOS is related to the clinical characteristics of PCOS.

Metabonomics reveals plasma metabolic changes and inflammatory marker in polycystic ovary syndrome patients.

Polycystic ovary syndrome (PCOS) is a common, clinically heterogeneous endocrine disorder affecting women of reproductive age, associated with endocrinopathy and metabolic abnormalities. Although some metabolic parameters have been investigated, very little information has been reported on the changes of small metabolites in biofluids. The aim of this study was to establish the metabolic profile of PCOS and compare it with that of controls. In this cross-sectional study of 34 women with PCOS and 36 controls, contents of small metabolites and lipids in plasma samples were measured using nuclear magnetic resonance (NMR)-based techniques and analyzed using multivariate statistical methods. Significant decrease (P < 0.05) in the levels of amino acids (leucine, isoleucine, methionine, glutamine, and arginine), citrate, choline, and glycerophosphocholine/phosphocholine (GPC/PC), and increase (P < 0.05) in the levels of lactate, dimethylamine (DMA), creatine, and N-acetyl glycoproteins were observed in PCOS patients compared with the controls. Subgroups of patients with obesity, metabolic syndrome, or hyperandrogenism exhibited greater metabolic deviations than their corresponding subgroups without these factors. PCOS patients have perturbations in amino acid metabolism, the tricarboxylic acid (TCA) cycle, and gut microflora, as well as mild disturbances in glucose and lipid metabolism. The elevated level of N-acetyl glycoproteins demonstrates the existence of low-grade chronic inflammation in PCOS patients.

[Study on synchronous correlation between fruit characteristic spectrum and the parameter of internal quality for Hamlin sweet orange fruit].

The purpose of our study is to study the relationship between the reflective spectrum of fruit and the internal quality of the orange fruit and find the suitable mark spectrum that can synchronously measure the several fruit quality index at same time to lay a foundation for the development of a rapid and nondestructive field fruit quality analysis technique by analyzing the visible-near infrared spectrum. Mature Hamlin orange (Citrus sinensis (L)cv. Hamlin sweet orange) fruits were individually mensurated for their reflective spectrum by using FieldSpec-HH spectrometer and for their contents of total soluble solid (TSS), citric acid and vitamin C (Vc) by chemistry analysis. The experiment results showed that the fruit reflectivity values (x) at 988 nm was significantly correlated to both TSS (y)(r = 0.387* *, y = 13. 957x + 5.405), TSS/acid ratio(y)(r = 0. 440* *, y = 75.120x + 37.256), and Vc(r = 0.309*). Both of TSS and Vc contends were positively correlated with the second derivatives of the reflective spectrum at 943 nm, with correlation coefficients of 0.339* and 0.355*. TSS/acid ratio was positively correlated only with the reciprocal log values of the reflective spectrum at 944 nm (r = 0.304*). The results in this study indicated that fruit quality indexes TSS, Vc and TSS/acid can be synchronously, rapidly and nondestructively field measured at the same time by the 988 or 429 nm reflective spectrum test and special regress equation operation.