Search

This study aimed to explore the temporal associations between maternal serum iodine concentration (SIC) and common pregnancy outcomes in Chinese women. Eligible singleton pregnant women aged 20–34 years were selected, and their fasting blood samples were collected during early (T1, n = 1101) and mid-pregnancy (T2, n = 403) for SIC testing by inductively coupled plasma mass spectrometry. Multivariable linear regression indicated that log10SIC at T1 (β = −0.082), T2 (β = −0.198), and their % change (β = −0.131) were inversely associated with gestational weight gain (GWG, all p < 0.05). Maternal log10SIC at both T1 (β = 0.077) and T2 (β = 0.105) were positively associated with the Apgar score at 1 min (both p < 0.05). Women in the third quartile (Q3) of SIC at T1 had a lower risk of small for gestational age (SGA, OR = 0.405, 95% CI: 0.198–0.829) compared with those in Q4. Restricted cubic spline regression suggested a U-shaped association between SIC and SGA risk, and SIC above 94 μg/L at T1 was the starting point for an increased risk of SGA. The risk of premature rupture of membrane (PROM) increased by 96% (OR = 1.960, 95% CI: 1.010–3.804) in Q4 compared to that in Q1. Our longitudinal data from an iodine-replete region of China indicated that high maternal SIC could restrict GWG and improve Apgar scores at delivery, but might increase the risk of SGA and PROM.

Background Surgery at the primary site of nasopharyngeal carcinoma (NPC) remains controversial. This study aimed to investigate the efficacy of surgery in patients with stage T1N0-3M0 NPC.Methods We reviewed 1803 patients with newly diagnosed stage T1N0-3M0 NPC from the Surveillance, Epidemiology, and End Results (SEER) database for the period 2004–2015. The Kaplan-Meier method and propensity score matching (PSM) analyses were used to compare cancer-specific survival (CSS) and overall survival (OS) between surgery groups and no-surgery groups. Restricted cubic spline regression (RCS) was used to assess the relationship between age and the hazard ratio of mortality. Independent risk factors associated with OS and CSS were identified by Cox ratio hazard regression analysis employing a backward stepwise method, which was used to develop prognostic nomograms for locoregional T1 NPC patients. The correlation index and calibration curves were used to assess the performance of the model. An online dynamic nomogram for CSS was also developed to assist the clinic in better using the predictive model.Results The study examined 1803 patients, of whom 242 received surgery, and the remaining 1561 did not receive surgery. Before PSM, Kaplan-Meier analysis showed that OS and CSS were significantly longer in the surgery group than in the non-surgery group (P 30 mm and primary site located in the anterior wall of the nasopharynx. Finally, COX regression analyses showed that age, sex, marital status, histological type, N stage, tumor size, radiotherapy, and surgery were independent prognostic factors for OS and CSS.Conclusions This study indicated that surgery significantly improved the OS and CSS of patients with stage T1N0-3M0 NPC. The nomograms we established have satisfactory performance and may provide a reference for the clinic. Further prospective large studies are still needed to validate the results.

The study aims to establish trimester-specific reference ranges for serum iodine (SI) in Chinese pregnant women and explore its associations with maternal and infantile thyroid function. Apparently healthy pregnant women were enrolled during their first antenatal visit. Fasting venous and spot urine samples were collected for determining serum and urinary iodine (UI) levels by a validated inductively coupled plasma mass spectrometry. Serum free triiodothyronine (FT3), free thyroxine (FT4), thyrotropin (TSH) and neonatal TSH levels were tested by electro-chemiluminescent assay. The reference ranges of SI were established by percentile method and reported as 2.5–97.5%. ROC analysis was applied to compare the discriminative ability of SI, UI and UI to urinary creatine ratio (UI /UCr) in early pregnancy for various thyroid conditions. The trimester-specific reference ranges of SI for Chinese pregnant women were 60.91-114.53 µg/L for the first trimester (T1, n = 1029), 54.57-103.42 µg/L for the second trimester (T2, n = 379) and 52.03–110.40 µg/L for the third trimester (T3, n = 455). Maternal SI at T1 but not UI and UI/UCr, was significantly correlated with FT3 (r = 0.393, P

High sensitivity and specificity imaging of miRNA in living cells plays an important role in understanding miRNA-related regulation and pathological research. Localized DNA circuits have shown good performance in reaction rate and sensitivity and have been proposed for sensitive imaging of miRNA in living cells. However, most reported localized DNA circuits have a high risk of derailment or a limited loading rate capacity, which hinder their further application. To solve these issues, we herein developed a domino-like localized cascade toehold assembly (LCTA) amplification-based DNA nanowire to achieve highly sensitive and highly specific imaging of miRNAs in living cells by using DNA nanowires as reactant delivery vehicles and confining both reactant probes in a compact space. The LCTA is constructed by interval hybridization of DNA double-stranded probe pairs to a DNA nanowire with multiplex footholds generated by alternating chain hybridization. Due to the localized effect, the LCTA showed high reaction kinetics and sensitivity, and the method could detect miRNAs as low as 51 pM. The LCTA was proven to be able to accurately distinguish the miRNA expression difference between normal cells and cancer cells. In particular, the developed LCTA could be used to construct an OR logic gate to simultaneously image the total amount of multiple miRNAs in living cells. We believe that the developed LCTA can be an effective intracellular nucleic acid imaging tool and can promote the development of nucleic acid-related clinical disease diagnosis and DNA logical sensors.

Traditional RCA methods face some drawbacks including limited specificity and amplification templates with sequence dependence. Herein, a universal RCA (URCA) strategy for label-free nucleic acid sensing with high specificity was proposed, which could be used for sensing of different nucleic acids without redesigning or synthesizing new amplification templates. The URCA strategy also showed high accuracy for miRNA analysis in practical samples.

Sensitive and specific imaging of microRNA (miRNA) in living cells is of great value for disease diagnosis and monitoring. Hybridization chain reaction (HCR) and DNAzyme-based methods have been considered as powerful tools for miRNA detection, with low efficient intracellular delivery and limited amplification efficiency. Herein, we propose a Hairpins@MnO

The highly sensitive detection of low-abundant apurinic/apyrimidinic endonuclease 1 (APE1) activity is of great significance for early diagnosis of disease and pathological research. Many methods for detecting APE1 based on isothermal nucleic acids amplification have been developed for improving its sensitivity. However, some of these methods have certain limitations, such as multiple reaction steps, narrow linear range, and complicated processes for fluorescent labeling. Herein, we develop a highly sensitive and label-free APE1 fluorescence detection method based on rolling circle amplification combined with G-quadruplex (RCA-G4). A hairpin probe (HP) labeled with the AP site can be recognized and cleaved by APE1, leading to the release of the primer sequence, which triggered RCA to produce long chain amplification products with a great amount of repeated sequences. The formed amplicon contains a series G-quadruplex structure, which can be combined with Thioflavin T (ThT) to produce fluorescence and achieve high sensitivity label-free detection of APE1. Benefit from the high amplification efficiency of RCA and the high fluorescence quantum yield of G-quadruplex/ThT, a detection limit as low as 1.52 × 10

PIWI-interacting RNAs (piRNAs) are a complex class of small non-coding RNAs which specifically interact with the PIWI protein to play important roles in germline development and somatic tissues. Aberrant expressions of piRNAs have been recently found in a variety of malignant tumors and associated with cancer hallmarks. However, current methods of analyzing piRNAs are limited to reverse transcription quantitative polymerase chain reaction and next generation sequencing. In this study, we have developed a universal catalytic hybridization assembly system (uniCHA) to quantify piRNAs as well as microRNAs. The system simply comprises two universal hairpin DNA strands and one starting hairpin DNA which can be tailored by a simple rule to bind different piRNA and miRNA targets. The uniCHA system was proved to be able to analyze various piRNAs and miRNAs at the same reaction condition with low leakage and high sensitivity of pM level. With this system, we have detected piR-651 and miR-1246 in 10

Sensitive, specific and rapid methods for detecting microRNAs (miRNAs) play critical roles in disease diagnosis and therapy. Enzyme-free amplification techniques based on DNAzyme assembly have recently been developed for the highly specific miRNA analysis. However, traditional DNAzyme-based assembly (free DNAzyme) amplifiers is mainly dependent on the target-induced split DNAzyme fragments to assemble into activated DNAzyme structures, which have made a compromise between the sensitivity and specificity due to the random diffusion of dissociative probes in a bulk solution with poor kinetics. Herein, based on a rationally designed DNA probe, we developed an intramolecular DNAzyme assembly (intra-DNAzyme) method to overcome these challenges. The miR-373 is used as model analyte for our current proof-of-concept experiments. Compared with the free-DNAzyme method, our method showed significantly improved analytical performance in terms of dynamic range, assay sensitivity and speed. This method can detect miR-373 specifically with a detection limit as low as 4.3 fM, which is about 83.7 times lower than the previous free-DNAzyme method. This intra-DNAzyme strategy would be of great value in both basic research and clinical diagnosis.

As the unique component of Dendrobium, dendrobine-type sesquiterpenoid alkaloids (DSAs) possess a variety of medicinal properties. It has been well documented that plant endophytes can in vitro synthesize secondary metabolites identical or similar to metabolites produced by their host plants. This study aimed to investigate the composition and distribution of endophytic bacteria of Dendrobium stems by Illumina MiSeq platform sequencing and cultivation-dependent methods and then to assess the potential for endophytic bacteria to produce DSAs. Results indicated that it was necessary to combine both cultivation-dependent and cultivation-independent methods to analyze the community structure of endophytic bacterial in plants comprehensively. The length of the Dendrobium stems influenced the endophytic bacterial community. The diversity and richness of endophytic bacteria in group J10_15cm of stems were the highest, which showed a significant difference from the other stem groups. However, there was no certain connection between the diversity and richness of endophytic bacteria and the content of dendrobine. It was most likely due to the influence of several specific endophytic bacteria genera, such as Sphingomonas and Rhodococcus. Athelia rolfsii, Myrothecium roridum, as pathogenic fungi, and Pectobacterium carotovorum subsp. actinidiae, as pathogenic bacteria of Dendrobium, were used to determine the antimicrobial activities. In these assays, six strains belonging to five genera showed antimicrobial activity against at least two phytopathogens. The strain BL-YJ10_15-29 (Paracoccus pueri THG-N2.35, 98.98%) showed the best antimicrobial activity against the three phytopathogens. In addition, 2 DSAs (6-hydroxydendrobine and nobilonine) were identified in the fermentation supernatant of the strain CM-YJ10_15-44 (Pseudomonas protegens CHA0, 99.24%), whereas the whole-genome analysis results further demonstrated that the precursors of the two DSAs [geranyl-PP and (E, E)-famesyl-PP] were synthesized mainly through the methyl-D-erythritol 4-phosphate pathway in this strain. This study provides new insight into the studies on the biosynthesis of DSAs and provides potential biocontrol bacteria.

Liquid biopsy is becoming an innovative tool in precision oncology owing to its noninvasive identification of biomarkers circulating in the body fluid at various time points for continuous and real-time analysis of disease progression. MicroRNAs in blood exosomes are identified as a new promising class of potential biomarkers for cancer diagnostics and prognostics. Conventional detection of blood exosomal microRNAs need multiple-step, complicated, costly, and time-consuming sample preparation of exosomes isolation and RNA extract, which affect the accuracy and reproducibility of analytical results. In this work, we set up an in situ quantitative analysis of human plasma exosomal miR-1246 by a probe of 2'-O-methyl and phosphorothioate modified molecular beacon. The probe has outstanding nuclease resistance in highly active RNase A/T1/I, which makes it stable for direct application in blood samples. With rapid rupture of exosomes membrane by Triton X-100, the probe can enter exosomes to specifically target miR-1246 exhibiting quantitative fluorescent signals. Using the output signals as a diagnostic marker, we differentiated 33 breast cancer patients from 37 healthy controls with 97.30% sensitivity and 93.94% specificity at the best cutoff. The blood biopsy is simple without extracting plasma exosomes and their nucleic acids content, time-saving in about 2 h of total analysis process, and microvolumes needed for plasma sample, suggesting its good potential to clinical application.

Background: Shenzhen implement classification management to prevent and control coronavirus disease 2019 (COVID-19) outbreak. Individuals with epidemic histories (came from Hubei and any other affected regions), but without symptoms were instructed to take home quarantine for 14 days and nucleic acid testing (NAT) for SARS-CoV-2. We described the infection status of the home-quarantined individuals, and effects of community control strategies in the three incubations after Wuhan closure in Shenzhen.Methods: This was a descriptive research, the sample size was 2,004 individuals based on multistage sampling during the pre-investigation. And the formal investigation expanded the sample size to 57,012 individuals based on pre-investigation. A single throat swab was collected from each individual for nucleic acid testing (NAT) by reverse transcription-polymerase chain reaction (RT-PCR). NAT was performed by a third-party institution. We collected information related to demographics, disease history, travel history, and personal protective measures before home quarantine, and monitored close-contact histories using the We Chat questionnaire.Results: The total infection rate of home-quarantined individuals was 0.12‰ (95% CI: 0.05‰–0.24‰) out of the total sample size of 59,016. The detection period for seven confirmed cases was primarily concentrated between February 8 and 18, 2020, which was during the second incubation period after Wuhan's closure. The home quarantined individuals with epidemic histories (came from Hubei and any other affected regions) were considered the high risk population during the first two incubations after Wuhan’s closure. No positive cases were detected from February 25 to March 5(the third incubation after Wuhan’s closure). The number of newly-confirmed cases per day was 0 for eight consecutive days from February 22 to 29 in Shenzhen.Conclusions: The community control strategies for home-quarantined individuals with epidemic histories (came from Hubei and any other affected regions), but without symptoms to take the NAT in the first two incubations is effective to control COVID-19. But it is not advocating for home-quarantined person to take the NAT since the third incubation.

AbstractBackground: To study the prevention and control strategies of coronavirus disease 2019 (COVID-19), and to analyze the infection of the home-quarantined individuals with epidemic histories (came from Hubei and any other affected regions), but without symptoms in the three incubations after Wuhan closure in Shenzhen.Methods: The sample size was 2,004 individuals based on multistage sampling during the pre-investigation. Based on the results of the pre-investigation, the formal investigation expanded the sample size to 57,012 individuals. A single throat swab was collected from each individual for nucleic acid testing (NAT) by reverse transcription-polymerase chain reaction (RT-PCR). NAT was performed by a third-party institution, BGI. We collected information related to demographics, disease history, travel history, and personal protective measures before home quarantine, and monitored close-contact histories using the We Chat questionnaire.Results: The total infection rate of home-quarantined individuals was 0.11% (95% CI: 0.05%–0.24%) out of the total sample size of 59,016. The detection period for seven confirmed cases was primarily concentrated between February 8 and 18, 2020, which was during the second incubation period after Wuhan's closure. The home quarantined individuals with epidemic histories (came from Hubei and any other affected regions) were considered the high risk population during the first two incubations after Wuhan’s closure. No positive cases were detected from February 25 to present (the third incubation after Wuhan’s closure). The number of newly-confirmed cases per day was 0 for 8 days from February 22 to 29 in Shenzhen. Thus, the strategies of prevention and control were effective.Conclusions: The strategies and policies were effective for the prevention and control of COVID-19. Additionally, the strategy of implementing NAT during the first two incubations for home-quarantined individuals with epidemic histories (came from Hubei and any other affected regions), but without symptoms, facilitated early detection, early reporting, early diagnosis, early quarantining, and early treatment. However, our findings do not support NAT for home quarantined persons during the third incubation after Wuhan’s closure to present.

Breast cancer is the second cause of cancer mortality in women globally. Early detection, treatment, and metastasis monitoring are of great importance to favorable prognosis. Although conventional diagnostic methods, such as breast X-ray mammography and image positioning biopsy, are accurate, they could cause radioactive or invasive damage to patients. Liquid biopsy as a noninvasive method is convenient for repeated sampling in clinical cancer prognostic, metastatic evaluation, and relapse monitoring. MicroRNAs encased in exosomes circulating in biofluids are promising candidate cancer biomarkers because of their cancer-specific expression profiles. Here, we report an in situ detection of microRNA-1246 (miR-1246) in human plasma exosomes as breast cancer biomarker by a nucleic acid functionalized Au nanoflare probe. Needing neither time-consuming and costly isolation of exosomes from the plasma sample nor transfection means, the Au nanoflare probe can directly enter the plasma exosomes to generate fluorescent signal quantitatively by specifically targeting miR-1246. Only 40 μL of plasma is needed to incubate 4 h with the probe, giving signal sensitive enough to distinguish samples of breast cancer to normal control. Using plasma miR-1246 level detected by our assay as a marker, we differentiated 46 breast cancer patients from 28 healthy controls with 100% sensitivity and 92.9% specificity at the best cutoff. This simple, accurate, sensitive, and cost-effective liquid biopsy by the Au nanoflare probe is potent to be developed as a noninvasive breast cancer diagnostic assay for clinical adaption.

Reactions of La(NO3)3·6H2O with the polar, tritopic quaternized carboxylate ligands N-carboxymethyl-3,5-dicarboxylpyridinium bromide (H3CmdcpBr) and N-(4-carboxybenzyl)-3,5-dicarboxylpyridinium bromide (H3CbdcpBr) afford two water-stable metal–organic frameworks (MOFs) of {[La4(Cmdcp)6(H2O)9]}n (1, 3D) and {[La2(Cbdcp)3(H2O)10]}n (2, 2D). MOFs 1 and 2 absorb the carboxyfluorescein (FAM)-tagged probe DNA (P-DNA) and quench the fluorescence of FAM via a photoinduced electron transfer (PET) process. The nonemissive P-DNA@MOF hybrids thus formed in turn function as sensing platforms to distinguish conservative linear, single-stranded RNA sequences of Sudan virus with high selectivity and low detection limits of 112 and 67 pM, respectively (at a signal-to-noise ratio of 3). These hybrids also exhibit high specificity and discriminate down to single-base mismatch RNA sequences.

We report herein five sensing platforms for the detection of five gastric cancer associated microRNAs (miRNAs). The sensing platforms are hybrids formed from a water-stable metal organic framework (MOF) {[Cu(dcbb)2(H2O)2]·10H2O}n (1, H2dcbbBr=1-(3,5-dicarboxybenzyl)-4,4'-bipyridinium bromide), respectively with five carboxyfluorescein (FAM) labeled probe single-stranded DNA (probe ss-DNA, denoted as P-DNA). Within the hybrid, MOF 1 tightly interacts with the P-DNA through electrostatic and/or π-stacking interactions and results in fluorescence quenching of FAM via a photo-induced electron transfer (PET) process. In the presence of the complementary target miRNAs miR-185, miR-20a, miR-92b, miR-25 and miR-210, which are expressed abnormally in the plasma of gastric carcinoma patients, P-DNA is released from the surface of MOF 1 ascribed to the stronger base pair matching, leading to the FAM fluorescence recovery. Each P-DNA@1 system is effective and reliable for the detection of its complementary target miRNA with the detection limits from 91 to 559pM, and is not interfered by other four miRNA sequences.

Coordination reaction of a known three-dimensional (3D) polymer precursor {Na3[Na9(Cbdcp)6(H2O)18]}n (A, Cbdcp = N-(4-carboxybenzyl)-(3,5-dicarboxyl)pyridinium) with Zn(NO3)2·6H2O in H2O or H2O/DMF at 100 °C and in the presence of aspirin, 5-fluorouracil (5-FU) as modulators, trans-1,2-bis(4-pyridyl)ethylene (bpe) or 1,2-bis(4-pyridyl)ethane (bpea) as ancillary ligands afforded six novel Zn(II)-based metal-organic frameworks (MOFs), that is, {[Zn(Cbdcp)(H2O)3]·H2O}n (1, 1D zigzag chain), {[Zn(HCbdcp)2]·H2O}n (2, 2D sheet), {[Zn(Cbdcp)(bpe)1/2]·2H2O}n (3, 3D polymer), {[Zn(Cbdcp)(bpe)1/2]·2H2O}n (4, 2D network), {[Zn(Cbdcp)(bpea)1/2]·2H2O}n (5, 3D polymer) and {[Zn(Cbdcp)(bpea)1/2]·2H2O}n (6, 2D network). Among them, compound 2 contains aromatic rings, positively charged pyridinium, Zn2+ cation centers and carboxylic acid groups lined up on the 2D sheet structure with a certain extended surface exposure. The unique structure of 2 facilitates effective association with carboxyfluorescein (FAM) labeled probe single stranded DNA (probe ss-DNA, delineates as P-DNA) to yield a P-DNA@2 system, and leads to fluorescence quenching of FAM via a photoinduced electron transfer process. The P-DNA@2 system is effective and reliable for the detection of human immunodeficiency virus 1 ds-DNA (HIV ds-DNA) sequences and capable of distinguishing complementary HIV ds-DNA from mismatched target sequences with the detection limit as low as 10 pM (S/N = 3).

A water stable one-dimensional (1D) ladder-shaped coordination polymer (CP) has been synthesized and exhibits a strong affinity to two fluorescein-tagged single-stranded probe DNAs (P-DNAs), giving a sensing platform of P-DNAs@1. Such a hybrid sensing platform is capable of simultaneous detection of breast cancer related microRNA-221 (miRNA-221) and miRNA-222 in a specific and synchronous manner, without observable cross-reactions, as supported by experimental evidences. The interaction mode and the electronic energy between CP 1 with nucleic acid were confirmed by molecular simulation and the universal force field (UFF).

Background Wound healing is a complex process that relies on growth factors and stimulation of angiogenesis. Tissue engineering materials composed of adipose-derived stem cells (ADSCs) and silk fibroin (SF)/chitosan (CS) may be able to solve this problem. The aim of this study was to investigate the wound-healing potential of ADSC-seeded SF/CS in streptozotocin-induced diabetic rats. Materials and methods Thirty-six male Sprague-Dawley rats were purchased and randomly assigned into 3 groups: a control group (no graft), a group treated with SF/CS film graft, and a group treated with ADSC-seeded SF/CS graft. The number of animals in each group was 12. Diabetes was induced by an intraperitoneal injection of streptozotocin. A cutaneous wound was incised at the dorsal region of all the experimental animals. The ADSCs were labeled with CM-Dil fluorescent staining. Wound healing was assessed for all animal groups by observing the rate of wound closure and hematoxylin and eosin staining. The expression of epidermal growth factor, transforming growth factor-β, and vascular endothelial growth factor at the wound sites was studied by enzyme-linked immunosorbent assay to evaluate the effect of growth factors secreted by ADSCs. The differentiation of ADSCs was analyzed by immunofluorescence staining. Results The ADSC-seeded SF/CS film treatment significantly increased the rates of wound closure in treated animals, and hence wound healing was drastically enhanced for ADSC-SF/CS treatment groups compared with control groups and SF/CS film treatment group. Histological observations showed the condition of wound healing. Enzyme-linked immunosorbent assay and immunofluorescence staining observations showed the secretion and differentiation of ADSCs, respectively. Conclusions Our analyses clearly suggested that it is feasible and effective to enhance wound healing in a diabetic rat model with ADSC-seeded SF/CS film.

Objective: Fluorine-18-fluorodeoxyglucose positron emission tomography (18F-FDG-PET) is a new technique for identifying different malignant tumors using different uptake values between tumor cells and normal tissues. Here we assessed the diagnostic accuracy of 18F-FDG-PET in patients with testicular cancer by pooling data of existing trials in a meta-analysis. Methods: PubMed/MEDLINE, Embase and Cochrane Central Trials databases were searched and studies published in English relating to the diagnostic value of FDG-PET for testicular cancer were collected. The summary receiver operating characteristic (SROC) curve was used to examine the FDG-PET accuracy. Results: A total of 16 studies which included 957 examinations in 807 patients (median age, 31.1 years) were analyzed. A meta-analysis was performed to combine the sensitivity and specificity and their 95% confidence intervals (CIs), from diagnostic odds ratio (DOR), positive likelihood ratios (PLR), negative likelihood ratio (NLR). SROC were derived to demonstrate the diagnostic accuracy of FDG-PET for testicular cancer. The pooled sensitivity and specificity were 0.75 (95% confidence interval (CI), 0.70-0.80) and 0.87 (95% CI, 0.84-0.89), respectively. The pooled DOR was 35.6 (95% CI, 12.9-98.3). The area under the curve (AUC) was 0.88. The pooled PLR and pooled NLR were 7.80 (95% CI, 3.73-16.3) and 0.31 (95% CI, 0.23-0.43), respectively. Conclusion: In patients with testicular cancer, 18F-FDG-PET demonstrated a high SROC area, and could be a potentially useful tool if combined with other imaging methods such as MRI and CT. Nevertheless, the literature focusing on the use of 18F-FDG-PET in this setting still remains limited.

This systematic review and meta-analysis assesses whether concurrent chemoradiotherapy is associated with improvement in survival outcomes for patients with locoregionally advanced nasopharyngeal carcinoma., Key Points Question Is induction chemotherapy or adjuvant chemotherapy associated with additional survival benefit in locoregionally advanced nasopharyngeal carcinoma? Findings In a systematic review and meta-analysis of 28 randomized clinical trials of 8036 patients, concurrent chemoradiotherapy was associated with substantial improvement in survival outcomes for patients with locoregionally advanced nasopharyngeal carcinoma. Survival benefit was also associated with the addition of induction chemotherapy but not adjuvant chemotherapy to concurrent chemoradiotherapy. Meaning For locoregionally advanced nasopharyngeal carcinoma, concurrent chemoradiotherapy should be recommended as the standard treatment strategy, with the addition of induction chemotherapy but not adjuvant chemotherapy., Importance The role of induction chemotherapy (IC) or adjuvant chemotherapy (AC) in the treatment of locoregionally advanced nasopharyngeal carcinoma (NPC) remains controversial. Objectives To update meta-analyses on the association of survival outcomes with IC and AC regimens in patients with locoregionally advanced NPC and assess whether the current evidence is conclusive by a trial sequential analysis (TSA) approach. Data Sources PubMed, Embase, and Web of Science were searched for articles published from inception until June 1, 2019. Study Selection Randomized clinical trials that assessed the efficacy of radiotherapy with or without chemotherapy among previously untreated patients and patients with nondistant metastatic NPC. Data Extraction and Synthesis Data were extracted by 2 investigators from each trial independently and synthesized by the 2 investigators. All trial results were combined and analyzed by a fixed- or random-effects model. Main Outcomes and Measures Overall survival (OS), progression-free survival (PFS), distant metastasis–free survival (DMFS), and locoregional recurrence-free survival (LRFS). Results A total of 8036 patients (median age, 46.5 years; 5872 [73.1%] male) from 28 randomized clinical trials were included in the analysis. Pooled analyses revealed that concurrent chemoradiotherapy (CCRT) was significantly associated with improved OS, PFS, DMFS, and LRFS compared with radiotherapy across all subgroups. The TSA confirmed the treatment outcomes of CCRT compared with radiotherapy. The additional IC regimen was associated with an improvement in OS (hazard ratio [HR], 0.84; 95% CI, 0.74-0.95), PFS (HR, 0.73; 95% CI, 0.64-0.84), DMFS (HR, 0.67; 95% CI, 0.59-0.78), and LRFS (HR, 0.74; 95% CI, 0.64-0.85). These findings were consistent in subgroup analyses of multicenter trials with sample sizes greater than 250, years of survival rate of 5 or greater, median follow-up longer than 5 years, or low risk of bias. However, the additional AC regimen was not associated with a survival benefit in OS (HR, 0.98; 95% CI, 0.78-1.23), PFS (HR, 0.86; 95% CI, 0.70-1.07), DMFS (HR, 0.84; 95% CI, 0.64-1.10), or LRFS (HR, 0.80, 95% CI, 0.59-1.09). The TSA provided sound evidence on the additional benefit of IC but not AC. Conclusions and Relevance These data suggest a significant association of survival outcomes with CCRT in patients with locoregionally advanced NPC. The addition of IC instead of AC could achieve survival benefits. The potential therapeutic gain of AC should be explored in the future.

In this report, the anti-hepatitis B virus (HBV) activity of dehydrocheilanthifoline (DHCH), a quaternary ammonium alkaloid isolated from the traditional Chinese medicine Corydalis saxicola Bunting (Papaveraceae), was determined in vitro. Following six days of treatment, DHCH efficiently suppressed the secretions of HBsAg and HBeAg in HepG2.2.15 cell cultures, with a half-maximal inhibitory concentration (IC50) of 15.84 and 17.12 μM, and with a therapeutic index (TI) of 7.32 and 6.77, respectively. Further studies revealed that DHCH reduced the levels of extracellular DNA, intracellular DNA and covalently closed circular DNA (cccDNA) of HBV in a dose-dependent and time-dependent manner, with IC50values of 15.08, 7.62 and 8.25 μM, respectively after six days of treatment. In contrast, the level of viral pre-genomic RNA (pgRNA) increased 6.13-fold after treatment with DHCH. Together, it was demonstrated for the first time that DHCH could significantly inhibit the replication of HBV, which warrants further studies on the antiviral mechanisms of DHCH, and suggests that it may be a promising candidate in the therapy of HBV infection.

Cabernet Sauvignon grafted onto seven rootstocks 188-08, 5BB, SO4, 3309C, 110R, 5C and 101-14M, with the own-rooted vines as control, were investigated to study the effects of different rootstocks on the growth, fruit quality and yield of Cabernet Sauvignon in Changli zone, Hebei Province, China. The results showed that Cabernet Sauvignon grafted on 5BB and 5C significantly increased the trunk diameter, and 5C significantly increased one-year-old shoot diameter. 188-08, 5BB and 5C as rootstock obviously improved berry soluble solid content, in addition 188- 08 and 5BB significantly increased berry reducing sugar content. The vines on 101-14M and 3309C significantly decreased berry titratable acid content. The rootstock 5C and 101-14M significantly raised grape skin phenol and anthocyanin contents, and rootstock 101-14M significantly increased tannin content in grape skin. Cabernet Sauvignon grafted onto 3309C, 110R, 5C and 101-14M obviously got higher yield per vine than own-rooted vines. Growing parameter, grape quality index and yield per vine grafted on seven rootstocks and own-rooted vine were synthetically evaluated by fuzzy evaluation method, and the synthetical effects of vine grafted on seven rootstocks were better than own-rooted vine, with the order of scores from high to low as 5C, 101-14M, 3309C, 5BB, 188-08, 110R and SO4 under Changli unique climate and environment conditions.

The purpose of study was to investigate the in vitro proliferation ability of PHA-induced CIK cells and traditionally prepared CIK cells, the effector cell level and its influence on killing activity to K562 cells, and to analyze the difference between them. The peripheral blood mononuclear cells(PBMNC) of healthy persons were isolated and divided into A and B group. The CIK cells in A group were obtained by using traditional culture method, the CIK cells in B group were prepared by PHA induction. During the cultivation, the cell survival rate and cell absolute value in the cell culture system were counted every 3 days. On day 15 of culture, the cell immunophenotype of 2 groups were detected by flow cytometry, and the ratios of CD3(+)CD56(+), CD3(+)CD8(+) and CD3(+)CD4(+) cells in total cell amount of culture system were accounted. Meantime, the killing activity to K562 cells in different effector-target ratios was detected by using CCK-8 kit between the 2 groups. The results showed that the method of preparing CIK by PHA induction promoted the cell proliferation more than that of the traditional method (P0.05), moreover, both the survival rate of cells in 2 groups was more than 90%. The CD3(+)CD8(+), CD3(+)CD56(+) cell ratio in 2 groups obviously increased. As compared with traditional method, the CD3(+)CD8(+) cell level in B group was enhanced (P0.05); but there were no statistical differences in increase of CD3(+)CD56(+) cell level and decrease of CD3(+)CD4(+) cell level between 2 groups. while the effector-target ratio is 5:1, 10:1, 20:1 and 40:1, the killing activity of PHA-induced CIK cells to K562 cells was more stronger than traditionally-prepared CIK cells (P0.05), moreover, along with increase of effector-target ratio, the difference of killing activity to K562 cells in 2 groups significantly increased. It is concluded that compared with traditional method for preparing CIK cells, the new way by PHA induction can increase the proliferation of CIK cells obviously, enhance the ratio of CD3(+)CD8(+) cells and strengthen the killing activity to the K562 cells. This new way provides a new source of CIK cells and reliable evidence for cyto-immune therapy of leukemia and other tumors.

To study and compare the anti-inflammatory effect and molecular mechanism of artemisinin and dihydroartemisinin.Mouse mononuclear macrophage RAW264.7 cells were stimulated to release inflammatory mediators such as TNF-alpha, IL-6 and NO, in order to assess the drugs' inhibitory effect on macrophage's release of above inflammatory mediators. The levels of TNF-alpha and IL-6 were determined by ELISA and the cytotoxicity was determined by MTT method. The protein expression of iNOS, COX-2 and beta-actin were tested by Western blot. The enzymatic activity of COX-2 was determined by colorimetric method.Dihydroartemisinin significantly inhibited LPS-induced release of TNF-alpha, IL-6 and NO from RAW264.7 in mice with the concentration range of 12.5 - 100 micromol x L(-1), and showed good dose dependence. Artemisinin only inhibited the IL-6 release to a certain extent.Dihydroartemisinin inhibits macrophages from releasing inflammatory factors TNF-alpha and IL-6 and inflammatory mediators NO by down-regulating iNOS protein. Artemisinin may help dihydroartemisinin to show its anti-inflammatory effect through metabolism.

To investigate whether the HIV-1 gp41 fusion peptide (FP) could affect the regulatory T cell (Treg) function activated by CD3 antibody.Murine CD4(+);CD25(+); Treg and CD4(+);CD25(-); effector T cells (Teff) were isolated from mice spleens by the immune magnetic beads. The splenocytes were treated with mitomycin C to obtain antigen presenting cells (APCs). CCK-8 assay and CFSE loading were employed to evaluate the effects of FP on the Treg inhibitory function via measuring Teff proliferation activated by CD3 antibody. In addition, IL-10 secretion of activated Treg was detected by ELISA. The distribution of FP and TCR on Treg surface was observed by laser scanning confocal microscope.Through the CCK-8 assay and flow cytometry, we found that Teff cells have significant proliferation stimulated by the CD3 antibodies. When Treg and Teff were co-cultured, the proliferation of Teff was significantly inhibited by Treg. When added with 25 μg/mL FP, the proliferation of Treg+Teff group and Teff group was not significantly affected, but when 5 μg/mL FP was added, the proliferation rate of Treg+Teff group was significantly lower than that of Teff group. IL-10 secretion was low when Treg were not activated, but it significantly increased by CD3 antibodies stimulation. When the concentration of FP was 25 μg/mL, IL-10 level significantly decreased, but 5 μg/mL FP did not significantly influence IL-10 secretion. Through the laser scanning confocal microscope, we found that T cell receptors (TCR) of non-activated Treg showed uniform distribution on the cell surface, and that FP and TCR had no common distribution. When Treg were stimulated by CD3 antibodies, the activated TCR formed half crescent, and FP and the activated TCR had common distribution on cell surface.Treg inhibitory function is significantly inhibited by 25 μg/mL FP in vitro, but 5 μg/mL FP does not affect it. This may be due to the suppression of IL-10 secretion and the influence of the signaling cross-talk between APC and TCR.

The aim of study was to investigate the effect of acute lymphoblastic leukemia (ALL) children bone marrow mesenchymal stem cells (MSC) on resistance of K562/A02 cells and its mechanism. MSC obtained from bone marrow of AL children were cultured and identified. The co-culture of MSC and K562/A02 and the culture of K562/A02 cell suspension alone was performed, of which 2 kinds of cells were treated with same concentration of adriamycin (ADM), and the rate of apoptosis was detected by flow cytometry, bcl-2 and bax of K562/A02 were detected by RT-PCR, while mdr1 gene level was detected by FQ-PCR. The results indicated that the MSC separation and proliferation were viable and steady. The apoptosis rate of the K562/A02 cells co-cultured with MSC was 1.97 ± 0.11%, while apoptosis rate of the K562/A02 cells cultured alone was 8.38 ± 0.29%, there was significant difference (p0.05). As compared with the K562/A02 cells cultured alone, the bcl-2 gene expression in K562/A02 cells co-cultured with MSC obviously increased; ratio of bcl-2/bax was obviously enhanced. The mdr1 gene level in K562/A02 co-cultured with MSC was no statistical different from K562/A02 cultured alone (p0.05), which suggested that adhesion co-cultured with MSC did not induce mdr1 expression higher than the culture of suspension. It is concluded that the MSC of ALL children can escape the leukemia cells from proapoptotic effect of drugs, the resistance of K562/A02 to ADM may be involved in enhancement of bcl-2 gene expression of K562/A02 cells co-cultured with MSC, but not in relation to mdr1 gene in K562/A02 cells themselves.

To develop an objective bioassay for quantitative detection of HIV-induced cell-cell fusion for screening HIV entry inhibitors.HL2/3 cells expressing HIV envelope proteins gp120/gp41, Tat, and other HIV proteins were co-cultured with HeLa-CD4-LTR-beta-gal cells expressing CD4 receptor and HIV LTR triggered reporter gene beta-galactosidase. The enzyme activities of beta-galactosidase were detected by a chromogenic substrate, chlorophenol red-beta-galactopyranoside (CPRG). Specific HIV entry inhibitors were used to validate the established detecting method.No syncytium was formed by mixing HL2/3 and HeLa-CD4-LTR-beta-gal cells. However, the membrane could be fused and the Tat expressed by HL2/3 cells could bind to HIV LTR on HeLa-CD4-LTR-beta-gal cells and trigger the expression of beta-galactosidase. CPRG allowed quantitative and sensitive detection of the activity of beta-galactosidase. Further studies showed that HIV entry inhibitors could inhibit the activity of beta-galactosidase in a dose-dependent manner.We have developed a simple, cheap, objective and quantitative non-infectious cell-cell fusion bioassay that can be used to screen for anti-HIV agents targeting the virus entry from natural and synthetic compound libraries.

To detect the serum anti-nuclear antibody (ANA) and interleukin-21 (IL-21) levels in patients infected with hepatitis C virus (HCV), and study their impact on the liver function of HCV patients.Enzyme-linked immunosorbent(ELISA) assays were used to detect the serum ANA and IL-21 levels in 96 cases of HCV-IgG positive sera, as well as 30 cases of HCV-IgG negative sera. Hitachi 7600 biochemical analyzer was used to test the ALT, AST and ALB of all sera samples.HCV patients have abnormal liver function significantly and their average IL-21 level was much lower than the control group. While the liver function of most ANA positive HCV patients was well-balanced. The IL-21 level of HCV patients with ALTor = 40 U/L or ALT40 U/L were markedly reduced.The production of ANA and IL-21 do help to control the progress of hepatitis and reduce the damage to liver cells in HCV patients.

To study the increasing incidence and the characteristics of Tsutsugamushi disease in the areas of Nan Peng Lie islands, Nan Ao island, Wan Shan archipelago, Nao Zhou island and Lei Zhou peninsula, located in the southern part of China and to develop strategies for preventive measures.Both epidemiological investigation, isolation and gene identification of Orientia tsutsugamushi, as well as pilot preventive measures were carried out.These islands belonged to the epidemic area of south subtropical zone of Tsutsugamushi disease. The main host was Rattus norvegicu and the overall rates of infection on Orientia tsutsugamushi were 22.78%-33.75%. The main biological vector was Leptotrombidium (Leptotrombidium) deliens and the rates of infection on Orientia tsutsugamushi were 40.00%-75.00%. 25 strains of Orientia tsutsugamushi had been isolated from Rattus norvegicu and Leptotrombidium (Leptotrombidium) deliens. Results showed that the isolated strains of Orientia tsutsugamushi were 15 Karp, 8 Kato, 2 Yonchon. Results from serological studies showed that the positive rate of anti-Orientia tsutsugamushi antibodies was high, in both residents and soldiers stationed in these islands. On these islands, rats and biological vectors were killed. Results showed that these measures had positive impact in reducing the incidence.Islands from the southern part of the country belonged to the epidemic area of Tsutsugamushi disease. People visiting this areas should be under protection.

Hepatitis B virus (HBV) DNA was detected from infants whose mothers were negative for all HBV markers and the fathers were HBV carrier, the homology of HBV sequence of fathers and fetus was high, and HBV mutations concentrated on some points, and the transmission of HBV from father to fetus was also identified in some reports. The present study aimed to study HBV transmission from father to infant.The study enrolled 16 pairs of fathers who were HBV carriers and infants whose mothers were negative for HBV markers. The infants had evidences for intrauterine HBV infection. The five HBV serum markers HBsAg, HBeAg, anti-HBe, anti-HBs, and anti-HBc were detected with ELISA. The positive results for HBsAg and/or HBeAg were regarded as markers of HBV infection. Amplification of HBV DNA was done using a nested PCR method. The first amplification was carried out using primer C1 (nt 2394-2370), and primer C3 (nt 1730-1754). The second amplification was carried out using primer C2 (nt 1955-1974) and primer C6 (nt 2348-2330). Both primers were designed to amplify the part of sequence coding for the hepatitis B C antigen. The size of the amplified fragment obtained by the nested PCR was expected to be 394 bp. The PCR products were electrophoresed on 1.5% agarose gels, which were then stained with ethidium bromide and observed with ultraviolet transillumination. When 394 bp specific band was detectable, the sample was designated positive. Then the positive samples were identified by dot blot. The second PCR products were extracted by phenol-chloroform and 70% ethanol precipitation, then resuspended in TE buffer (pH8.0), and used as the template for cloning. The template was connected into pGEM-T vector by ligase. The ligated products were cloned into fresh competent JM109 cells, and incubated for 90 minutes at 37 degrees C on roller drum. Finally several dilutions were plated on plates containing ampicillin, X-Gal and IPTG, and incubated at 37 degrees C overnight. The white colony on plates was used for identification by the nested PCR with the above primers. When the 394 bp band was detectable by electrophoresis of PCR products in 1.5% agarose gels, the colony was designated positive; a positive colony was incubated in LB medium for 8 to 12 hrs, then plasmid was extracted using the Wizard Plus SV Minipreps DNA Purification System Kit (Promega). The purified plasmid was sent to Beijing Saibaisheng Company for sequencing. The homology of HBV C nt 2022-2301 sequence was compared between fathers and infants.The homology of HBV C nt 2022-2301 sequence were 99% - 100% in 16 pairs of fathers and infants. The results were referred to the published sequence of HBV adw/adr clones, and the nucleic acid databases were searched for homology by using BLAST tool on Internet. HBV of the sixteen pairs of father/infant was closely related to the Japan strain (Genebank accession number AF121249), but there were still 17 more mutations at nucleotide positions 2029, 2034, 2044, 2059, 2078, 2095, 2104, 2154, 2161, 2169, 2189, 2201, 2233, 2251, 2284, 2288, 2293. Moreover the mutations at positions 2189, 2288 resulted in the substitution of the encoded amino acid (corresponding to amino acid positions 97 and 130, respectively), the other mutations at the position were nonphenotypic. The mutation of 2189, 2288 nucleotide of HBV C gene caused 97, 130 amino acid substitution for isoleucine to leucine and proline to threonine. The mutation of 2189, 2288 nucleotide of HBV C gene were detected in 6 (37.5%) of 16 pairs of fathers and infants.The HBV transmission from father to infants did exist. The main HBV C gene mutation strains also existed in the transmission.

To investigate the mechanism for vertical transmission of hepatitis C virus (HCV) infection.This study included 6 pairs of mothers and fetuses who were positive for anti-HCV antibody and HCV RNA, and 2 pairs of fathers and fetus in whom the father was positive for anti-HCV and HCV RNA while the mother was negative. HCV genotypes were detected by PCR and the nucleotide sequences of HCV genome spanning the core and envelope gene region (HCV C/E1) determined and compared.The HCV genotypes of the mothers and fetus were consistent. Five mother-fetus pairs were of genotype II, 1 of genotype III, with high homology of HCV between the mothers and their own fetuses. In contrast, in the two father-fetus pairs, no HCV RNA was detected in the fetuses in spite of the positivity of the fathers.Vertical transmission of HCV infection from mothers to fetus is a clinical entity.