Efficacy and safety of corticosteroid therapy in patients with cardiac arrest: A meta-analysis of randomized controlled trials.

BACKGROUND: The clinical benefits of steroid therapy during cardiac arrest (CA) are unclear. Several recent clinical trials have shown that administering corticosteroid therapy during CA may improve patient outcomes. The purpose of the present study was to determine whether providing corticosteroids improves outcomes for patients following CA. METHODS: We searched the PubMed, Embase, Cochrane Library, Web of Science and CNKI databases for randomized controlled trials comparing corticosteroid therapy to placebo during CA. RESULTS: Eleven relevant studies involving a total of 2273 patients were included in the meta-analysis. The statistical analysis showed that corticosteroid treatment during CA was significantly associated with an increased rate of sustained return of spontaneous circulation (ROSC) (OR: 2.05, 95% CI: 1.24 to 3.37, P < 0.01). Corticosteroid treatment during CA did not show a significant benefit in favorable neurological outcomes (OR: 1.13, 95% CI: 0.81 to 1.58, P = 0.49) or overall survival rate at hospital discharge (OR: 1.29, 95% CI: 0.74 to 2.26, P = 0.38). However, in the subgroup analysis, we found that patients had a significantly increased survival rate and ROSC if the dose of corticosteroid therapy above 100 mg methylprednisolone. The statistical analysis revealed no significant differences in adverse events. CONCLUSION: High-dose corticosteroid treatment (above 100 mg methylprednisolone) is associated with better overall survival rate at hospital discharge and ROSC outcomes. However, there is uncertainty regarding whether this treatment results in a benefit or harm to the favorable neurological outcomes at hospital discharge.

m6A-activated BACH1 exacerbates ferroptosis by epigenetic suppression HSPB1 in severe acute pancreatitis.

Severe acute pancreatitis (SAP) is characterized by acute inflammation of the pancreas. The transcription factor BTB and CNC homology 1 (BACH1) has been implicated in various biological processes, including oxidative stress, apoptosis, and cell cycle regulation. However, its involvement in the pathogenesis of SAP remains relatively understudied. In the present work, our data demonstrated that BACH1 level was significantly increased in SAP patients, cellular, and animal models, while heat shock protein B1 (HSPB1) expression was weakened. Mechanistic assays validated that BACH1 acted as a transcriptional inhibitor of HSPB1. Moreover, HPDE6-C7 cells were stimulated with cerulein (Cer) and LPS to mimic the pathological stages of SAP in vitro. Depletion of BACH1 remarkably improved cell survival and alleviated the oxidative stress, ferroptosis, and inflammatory responses in SAP cell models. However, these changes were dramatically reversed upon co-inhibition of HSPB1. Animal findings confirmed that loss of BACH1 decreased pancreatic injury, inflammatory responses, and ferroptosis, but these effects were weakened by HSPB1 silence. Overall, these findings elucidate that the overexpression of BACH1 favors the ferroptosis and inflammation by transcriptionally inhibiting HSBP1, thereby exacerbating SAP progression.

Coherent Modulation of the Aggregation Behavior and Intramolecular Charge Transfer in Small Molecule Probes for Sensitive and Long-term Nerve Agent Monitoring.

Aggregation-induced emission (AIE) shows promising performance in chemical sensing relying on the change of the emission behavior of the probe molecule monomers to the aggregated product. However, whether the response contrast could be further boosted by utilizing the emission property of the aggregated probe and the aggregated product remains a big challenge. Here, an exciting AIE probe regulation strategy was proposed by coherently modulating the aggregation behavior and the intramolecular charge transfer (ICT) property of the probes and thus an aggregated-to-aggregated colorimetric-fluorescent dual-mode detection was achieved. The blue emissive film obtained with the optimal AIE probe has been proven to be effective to recognize the vapor of nerve agent analog DCP in air by emitting a sharp green fluorescence. In addition, a porous polymer-based wet sensing chip loaded with the probe enables the immediate response to DCP vapor with a limit of detection (LOD) of 1.7 ppb, and it was further integrated into a wearable watch device for long-term monitoring of DCP vapor up to two weeks. We expect the present probe design strategy would greatly deepen the AIE-based science and provide new insights for long-term monitoring sensors toward trace hazardous substances.

Characterization of a Tigecycline-Resistant and blaCTX-M-Bearing Klebsiella pneumoniae Strain from a Peacock in a Chinese Zoo.

In Chinese zoos, there are usually specially designed bird parks, similar to petting zoos, that allow children and adults to interact with diverse birds. However, such behaviors present a risk for the transmission of zoonotic pathogens. Recently, we isolated eight strains of Klebsiella pneumoniae and identified two blaCTX-M-positive strains from 110 birds, including parrots, peacocks, and ostriches, using anal or nasal swabs in a bird park of a zoo in China. There, K. pneumoniae LYS105A was obtained from a diseased peacock with chronic respiratory diseases by a nasal swab, which harbored the blaCTX-M-3 gene and exhibited resistance to amoxicillin, cefotaxime, gentamicin, oxytetracycline, doxycycline, tigecycline, florfenicol, and enrofloxacin. According to an analysis by whole-genome sequencing, K. pneumoniae LYS105A belongs to serotype ST859 (sequence type 859)-K19 (capsular serotype 19) and contains two plasmids, of which pLYS105A-2 can be transferred by electrotransformation and harbors numerous resistance genes such as blaCTX-M-3, aac(6')-Ib-cr5, and qnrB91. The above-mentioned genes are located in a novel mobile composite transposon, Tn7131, which makes horizontal transfer more flexible. Although no known genes were identified in the chromosome, a significant increase in SoxS upregulated the expression levels of phoPQ, acrEF-tolC, and oqxAB, which contributed to strain LYS105A acquiring resistance to tigecycline (MIC = 4 mg/L) and intermediate resistance to colistin (MIC = 2 mg/L). Altogether, our findings show that bird parks in zoos may act as important vehicles for the spread of multidrug-resistant bacteria from birds to humans and vice versa. IMPORTANCE A multidrug-resistant ST859-K19 K. pneumoniae strain, LYS105A, was obtained from a diseased peacock in a Chinese zoo. In addition, multiple resistance genes such as blaCTX-M-3, aac(6')-Ib-cr5, and qnrB91 were located in a novel composite transposon, Tn7131, of a mobile plasmid, implying that most of the resistance genes in strain LYS105A can be moved easily via horizontal gene transfer. Meanwhile, an increase in SoxS can further positively regulate the expression of phoPQ, acrEF-tolC, and oqxAB, which is the key factor for strain LYS105A to develop resistance to tigecycline and colistin. Taken together, these findings enrich our understanding of the horizontal cross-species spread of drug resistance genes, which will help us curb the development of bacterial resistance.

A TD-DFT study of a class of D-π-A fluorescent probes for detection of typical oxidants.

A deep understanding of the fluorescence response mechanisms is the foundation for design-oriented strategies for D-π-A probes for trace hazardous chemicals. Here, from the perspective of electronegativity regulation of the π-bridge recognition site, an electron-donation modulation strategy involving various comprehensive evaluations of the optical and chemical properties is proposed through a series of theoretical analyses. Due to the preferential combined interaction between the π-bridge recognition site and MnO4-, high electrophilic reactivity and feasible chemical reaction energy barrier, a high-performance filter paper chip and hydrogel chip for the detection of aqueous and air-suspended environmental KMnO4 was achieved. We expect the present modulation strategy will facilitate efficient fluorescent probe design and provide a universal methodology for the exploration of functional D-π-A molecules.

D-π-A Dual-Mode Probe Design for the Detection of nM-Level Typical Oxidants.

Although a set of functional molecules with the D-π-A structure has been explored as optical probes for the detection of target analytes, it remains a great challenge to elaborately design a single probe for distinguishing different analytes by their intrinsic oxidation or reduction capabilities and thus to generate distinct optical responses. Here, a unique TCF-based probe (DMA-CN) containing two unsaturated double bonds in the π-conjugation bridge and TCF with different reaction activities that could be cut off by KMnO4 and NaClO in varying degrees was developed, causing remarkably distinguishable responses for both fluorescence and colorimetric channels to discriminate KMnO4 and NaClO from each other. The fluorescence and colorimetric limits of detection (LODs) of the proposed DMA-CN toward KMnO4 were calculated as 60 and 91 nM, respectively, while those for NaClO were 13.3 and 214 nM, and all the optical signal change can be observed within 1 s with good specificity. Based on the proposed probe design strategy, a well-fabricated test strip was proven to be promising for the rapid, in-field detection and risk management. We expect that the present probe design methodology would provide a powerful strategy for efficient probe exploration, especially for discriminating the substances with similar oxidizing properties.

Solvation effect enabled visualized discrimination of multiple metal ions.

Highly efficient detection of environmental residual potentially toxic species is of concern worldwide as their presence in an excessive amount would greatly endanger the health of human beings as well as environmental sustainability. The solvation effect is a critical factor to be considered for understanding chemical reaction progress as well as the photophysical behaviors of substances and thus is promising for visualized detection of metal ions. Herein, by applying 5-amino-1,10-phenanthroline (APT) as the optical probe, a sensing strategy was proposed based on the solvation effect modulated complexation of APT towards different metal ions to achieve the visualized discrimination of four critical ions (Cu(II), Zn(II), Cd(II), and Al(III)). How the crucial intrinsic properties of the solvent (e.g., polarity, solvent free energy, and electrostatic potential) influenced the complexation and the product emission was clarified, and the detection performances were systematically evaluated with detection limits as low as the nM level and good recognition selectivity. Furthermore, a portable sensing chip was developed with potential for highly efficient analysis in complicated scenes; thus, this strategy offers a new insight into determining multiple metal ions or other critical substances upon solvation manipulation.

Hereditary neuropathy with liability to pressure palsy: an investigation in a rare and large Chinese family.

BACKGROUND: Hereditary neuropathy with liability to pressure palsy (HNPP), mainly associated with the peripheral myelin protein 22 (PMP22) gene, is generally an autosomal-dominant inherited peripheral neuropathy. The present large family including four generations provides an exciting opportunity to gain important insights into HNPP in China. PATIENTS AND METHODS: A large 43-member family with ten members suspected to be affected by HNPP was studied. Neurologic examinations, electrophysiological and neuropathological studies and molecular genetic testing were used for these kindred. RESULTS: Clinically, the proband had limb hyposthenia and atrophy, and his mother showed declined tendon reflexes in the right lower limb. Electrophysiologically, sensory and motor nerve conduction velocities were generalized reduced. Sural nerve biopsy for the proband showed focal thickesning of the myelin sheaths. Furthermore, real-time quantitative PCR demonstrated that the PMP22 gene has a higher Ct value than reference gene in all suspected patients. CONCLUSIONS: These results indicated that the family is indeed a rare and large pedigree of HNPP caused by the deletion of PMP22 gene. Given that the suspected patient in the fourth generation is absent, this family is still worthy of further follow-up study.

Associations between rs3480 and rs16835198 gene polymorphisms of FNDC5 with type 2 diabetes mellitus susceptibility: a meta-analysis.

Background: FNDC5 is a novel and important player in energy regulation related to glucose metabolism and insulin levels. Thus, it may affect the incidence of type 2 diabetes mellitus (T2DM). Nevertheless, the association between FNDC5 single nucleotide polymorphisms (SNPs) and susceptibility to T2DM remains unclear. The aim of this meta-analysis was to explore whether the SNPs, rs3480 and rs16835198, are associated with the risk of T2DM. Methods: Studies published before February 1st, 2022 were screened to identify the included studies. R software was also applied for calculation of odds ratio (OR), 95% confidence interval (95% CI), heterogeneity, and sensitivity analysis. Results: Seven studies for rs3480 (involving 5475 patients with T2DM and 4855 healthy controls) and five studies for rs16835198 (involving 4217 patients with T2DM and 4019 healthy controls) were included in this meta-analysis. The results revealed a statistically significant association of rs3480 with T2DM under homozygote (GG vs AA: OR = 1.76, 95% CI = 1.31-2.37, P = 0.0002, I2 = 59%) genetic model. However, there was no statistically significant correlation between rs16835198 and susceptibility to T2DM under allelic (G vs T: OR = 1.33, 95% CI = 0.94-1.89, P = 0.11, I2 = 84%), heterozygote (GT vs TT: OR = 1.17, 95% CI = 0.80-1.69, P = 0.42, I2 = 71%), homozygote (GG vs TT: OR = 1.35, 95% CI = 0.95-1.94, P = 0.10, I2 = 62%), recessive (GG+GT vs TT: OR = 1.25, 95% CI = 0.88-1.79, P = 0.22, I2 = 72%), and dominant (GG vs GT+GG: OR = 1.20, 95% CI = 0.96-1.50, P = 0.11, I2 = 46%) genetic models. Conclusions: The present meta-analysis revealed that rs3480 in FNDC5 is significantly associated with susceptibility to T2DM, while rs16835198 does not show such an association.

Polyethyleneimine-capped copper nanoclusters for detection and discrimination of 2,4,6-trinitrotoluene and 2,4,6-trinitrophenol.

The detection and discrimination of 2,4,6-trinitrotoluene (TNT) and 2,4,6-trinitrophenol (TNP) from analogues are of great importance to global security and are full of challenges in the field of trace sensing. Here, benefitting from the strong electrophilicity of TNT, a sensing strategy is established by synthesizing polyethyleneimine capped copper nanoclusters (PEI-Cu NCs) with abundant -NH2 groups. By carefully controlling the size and structure of PEI-Cu NCs, Förster resonance energy transfer (FRET) from PEI-Cu NCs to the Meisenheimer complex occurs resulting from their spectral overlap when detecting TNT, while, due to the energy level match of TNP with PEI-Cu NCs, as well as the strong affinity between its -OH and -NH2 in PEI-Cu NCs, photo-induced electron transfer (PET) is feasibly expected. As a result, TNT and TNP could be detected from 26 types of analogues and cations with a limit of detection (LOD) of 26.57 and 12.82 nM, respectively. Besides, owing to the brown color of the Meisenheimer complex, the discrimination of TNT and TNP could be additionally realized by colorimetric detection. We expect that the proposed methodology would not only shine light on the detection and discrimination of TNT and TNP that mitigate against public security concerns, but also pave a way for the deep understanding of FRET and PET related fluorescence quenching mechanisms from the aspect of controllable sensing material design and synthesis.

Toll-like receptor 4/nuclear factor-kappa B pathway is involved in myocardial injury in a rat chronic stress model.

Chronic stress is considered to predispose to various cardiovascular events such as coronary artery disease, hypertension, and even heart failure. In this study, rats were exposed to stress for 1 day, 1, 2, 3, and 4 weeks to establish a chronic stress model. A specific toll-like receptor 4 (TLR4) antagonist eritoran was used to block the activity of TLR4. On the second day after the last stress exposure, the animals were killed. The expression of TLR4 mRNA and nuclear factor-kappa B (NF-κB) DNA-binding activity in the myocardium were measured using reverse transcriptase polymerase chain reaction and electrophoretic mobility shift assay. The proinflammatory cytokines such as tumor necrosis factor (TNF)-α and interleukin (IL-6) in myocardium were assayed by enzyme-linked immunosorbent assay. Myocardial injury was evident after chronic stress for 2 weeks. The TLR4 mRNA expression reached a peak after stress for 1 week. It was sustained at a stable level after stress exposure for 3 weeks and was restored to a nearly normal level in the fourth week. NF-κB DNA-binding activity was significantly enhanced after the stress for 1 day and markedly enhanced again after a 2-week stress exposure. It was weakened and reached a normal level after stress exposure for 4 weeks. The levels of TNF-α and IL-6 gradually increased and reached peaks after stress for 4 weeks. Meanwhile, eritoran significantly decreased the TLR4 mRNA expression and NF-κB activity in rats from the 2-week stress group. However, it did not downregulate the levels of TNF-α and IL-6. Importantly, it significantly improved the myocardial injury induced by the chronic stress. In conclusion, TLR4/NF-κB participates in myocardial injury during chronic stress.

Analysis of clinical features of 231 cases with pernicious placenta previa: A retrospective cohort study.

ABSTRACT: Pernicious placenta previa (PEPP) is a severe complication of late pregnancy, which might result in adverse maternal-fetal outcome. To explore the application value of placenta accreta score (PAS) for PEPP and its association with maternal-fetal outcome.In this retrospective cohort study, the clinical data of PEPP patients were analyzed. According to the ultrasonic PAS, patients were grouped into 3 groups: scores ≤5, a scores between 6 and 9, and scores ≥10. The clinical data, intraoperative and postoperative outcomes were collected. Receiver operating characteristic (ROC) curves were used to evaluate the performance of PAS in disease severity evaluation. Multivariate logistic and linear regression analysis were performed to assess associations of PAS with intraoperative and postoperative outcomes.A total of 231 patients were enrolled. There were significant differences in intraoperative, postoperative and neonatal outcomes, such as operation time, bladder repair, ICU admission, postoperative hospitalization days, operation complications, Apgar score of newborns in 1 minute and premature delivery among the 3 groups (all P < .05), while the worst outcomes were found in those with a score ≥ 10 (all P < .05). According to ROC curves, scores <5.5, between 5.5 and 7.5, and >7.5 indicated placenta accreta, placenta increta and placenta percreta, respectively. PAS was independently associated with longer time of operation, surgical complications, intraoperative bleeding volume, and postoperative hospitalization days (all P < .05).Placenta accreta score might help with PEPP subtype diagnosis and predict the maternal-fetal outcome of PEPP patients.

Doxazosin attenuates renal matrix remodeling mediated by anti-α1-adrenergic receptor antibody in a rat model of diabetes mellitus.

Diabetic nephropathy is a major complication of diabetes mellitus (DM). Recent studies suggest that immunological mechanisms have a key role in the pathogenesis of DM, therefore these mechanisms may be important targets for diabetes therapy. The present study evaluated the effects of anti-α1-adrenergic receptor antibody (α1-R Ab) mediation and doxazosin treatment in a rat model of DM. It was observed that levels of 24-h urinary protein, serum creatinine and transforming growth factor-ß1 in DM were significantly increased after α1-R Ab mediation (all P<0.05). In addition, electron microscopy identified severe damage in the renal tissue microstructures of DM rats following α1-R Ab mediation, while only mild abnormalities were observed in that of healthy rats mediated with α1-R Ab and of untreated DM rats. No marked abnormalities were observed in the renal tissue of healthy blank controls. Furthermore, in DM rats treated with α1-R Ab mediation + doxazosin intervention, the expression of TGF-ß1 significantly decreased, and renal functions and renal matrix remodeling were significantly improved, relative to untreated DM controls (P<0.01). These results suggest that α1-R Ab may be involved in renal matrix remodeling during DM, and that kidney protection during DM may be achieved through treatment with corresponding receptor antagonists.

Predomination and new genotypes of Enterocytozoon bieneusi in captive nonhuman primates in zoos in China: high genetic diversity and zoonotic significance.

To appreciate the genetic diversity and zoonotic implications of Enterocytozoon bieneusi in nonhuman primates (NHPs) in zoos, we genotyped E. bieneusi in captive NHPs in seven zoos located at six major cities in China, using ribosomal internal transcribed spacer (ITS)-based PCR and sequence analyses. A total of 496 fecal specimens from 36 NHP species under nine families were analyzed and E. bieneusi was detected in 148 (29.8%) specimens of 25 NHP species from six families, including Cercopithecidae (28.7%), Cebidae (38.0%), Aotidae (75.0%), Lemuridae (26.0%), Hylobatidae (50.0%) and Hominidae (16.2%) (P = 0.0605). The infection rates were 29.0%, 15.2%, 18.2%, 37.3%, 29.2%, 37.7% and 44.8% in Shijiazhuang Zoo, Wuhan Zoo, Taiyuan Zoo, Changsha Wild Animal Zoo, Beijing Zoo, Shanghai Zoo and Shanghai Wild Animal Park, respectively (P = 0.0146). A total of 25 ITS genotypes were found: 14 known (D, O, EbpC, EbpA, Type IV, Henan-IV, BEB6, BEB4, Peru8, PigEBITS5, EbpD, CM1, CM4 and CS-1) and 11 new (CM8 to CM18). Genotype D was the most prevalent one (40/148), followed by CM4 (20/148), CM1 (15/148), O (13/148), CM16 (13/148), EbpC (11/148). Of them, genotypes D, EbpC, CM4 and O were widely distributed in NHPs (seen in 9 to 12 species) whereas genotypes CM1 and CM16 were restricted to one to three NHP species. In phylogenetic analysis, 20 genotypes (121/148, 81.8%), excluding genotypes BEB4, BEB6, CM9, CM4 and CM18, belonged to group 1 with zoonotic potential. New genotype CM9 clustered in group 2 with BEB4 and BEB6. The remaining two genotypes CM4 and CM18 formed new cluster (group 9) in between two other genotypic clusters found in primates. The findings of high diversity in E. bieneusi genotypes and their zoonotic potentiality concluded the importance of captive NHPs as reservoir hosts for human microsporidiosis.

Clinical characteristics of 66 patients with novel coronavirus (2019-nCoV) -infected pneumonia (NCIP) in Enshi, Hubei / 中华急诊医学杂志

Objective:To describe the epidemiological and clinical characteristics of patients with novel coronavirus (2019-nCoV) -infected pneumonia in Enshi, Hubei, and to improve the awareness of the disease, which is key for surveillance and control measures of 2019-nCoV pneumonia in the region.Methods:The first 66 laboratory-confirmed 2019-nCoV patients in Enshi between January 23, 2020 and February 1, 2020 were included. Their epidemiological data, demographic data, clinical data and therapeutic effect were retrospectively analyzed.Results:Among the first 66 patients with confirmed 2019-nCoV, patients had an average age of 46±9 years, 35 (53.0%) patients were male and 31 (47.0%) female; 38 (57.6%) patients were Han, 18 (27.3%) were Tujia, and 10 (15.1%) were Miao. Thirty-seven (56%) patients had chronic diseases such as coronary heart disease, chronic bronchus, inflammation, diabetes, hypertension, hypothyroidism, and rheumatoid arthritis. All the patients had a history of exposure or indirect exposure in Wuhan epidemic area, mainly history of Wuhan sojourner. The mean incubation period varied from 2.5-16 days, with the 95th percentile of the distribution at 7 days. The main clinical manifestation were fever [66 (100%) patients], dry cough [56 (84.8%)], chest tightness [21 (31.8%)], shortness of breath [8 (12.1%)], massive fatigue [23 (34.8%)], muscle ache [6 (9.1%)], headache [4 (6.1%)], sore throat [13 (19.7%) ], rhinorrhoea [11 (16.7%) ], and diarrhea [ 5 (7.6%)]. Six (9.1%) critically ill patients and 4 (6.1%) critically ill patients had dyspnea. White blood cell counts were normal or decreased in 51 (77.3%) patients, and increased in 15 (22.7%). Lymphocyte counts were reduced or normal in 58 (87.9%) patients, and increased in 8 (12.1%) patients. Most patients had elevated CRP and erythrocyte sedimentation, and some patients had normal ranges of liver function, renal function, and electrolytes. Ten severely ill patients had elevated liver enzymes and cardiac enzymes (especially K and CKMB). There were significant differences in inflammatory markers among patients of different nationalities. Arterial blood gas analysis in 56 (84.8%) mild patients showed no significant abnormalities. Chest CT scans of 66 patients showed single or multiple small patchy shadows and interstitial changes, which were evident in the extrapulmonary band. All patients were hospitalized and isolated for treatment. Of the 66 patients received oxygen therapy, most of them received antiviral therapy (abidol/Kaletra, 0.2 g po tid/2 pills po bid) and interferon (aerosol inhalation), few patient received antibacterial therapy (Tanreqing, 20 mL ivgtt qd) and glucocorticoid therapy (methylprednisolone, 40 mg ivgtt bid), and supplemented by traditional Chinese medicine treatment. Six (9.1%) severely ill patients and 4 (6.1%) critically ill patients treated with non-invasive or invasive ventilator.Conclusions:The patients in this study are all imported cases. Most patients have mild clinical symptoms. The lungs show single or multiple small patchy shadows and interstitial changes, which are obvious with extrapulmonary bands. Seek medical treatment as soon as possible, and the above-mentioned symptomatic treatment scheme is effective. The overall morbidity and mortality in this region are lower than those in other parts of the province, and the mortality and critical illness rate of ethnic minority patients are lower than those of Han patients. There are significant differences in inflammatory markers such as leukocytes and lymphocytes among patients of different nationalities.

Multi-locus analysis of Giardia duodenalis from nonhuman primates kept in zoos in China: geographical segregation and host-adaptation of assemblage B isolates.

Only a few studies based on single locus characterization have been conducted on the molecular epidemiology of Giardia duodenalis in nonhuman primates (NHPs). The present study was conducted to examine the occurrence and genotype identity of G. duodenalis in NHPs based on multi-locus analysis of the small-subunit ribosomal RNA (SSU rRNA), triose phosphate isomerase (tpi), glutamate dehydrogenase (gdh), and beta-giardin (bg) genes. Fecal specimens were collected from 496 animals of 36 NHP species kept in seven zoos in China and screened for G. duodenalis by tpi-based PCR. G. duodenalis was detected in 92 (18.6%) specimens from 18 NHP species, belonging to assemblage A (n=4) and B (n=88). In positive NHP species, the infection rates ranged from 4.8% to 100%. In tpi sequence analysis, the assemblage A included subtypes A1, A2 and one novel subtype. Multi-locus analysis of the tpi, gdh, and bg genes detected 11 (8 known and 3 new), 6 (3 known and 3 new) and 9 (2 known and 7 new) subtypes in 88, 47 and 35 isolates in assemblage B, respectively. Thirty-two assemblage B isolates with data at all three loci yielded 15 multi-locus genotypes (MLGs), including 2 known and 13 new MLGs. Phylogenetic analysis of concatenated sequences of assemblage B showed that MLGs found here were genetically different from those of humans, NHPs, rabbit and guinea pig in Italy and Sweden. It further indicated that assemblage B isolates in ring-tailed lemurs and squirrel monkeys might be genetically different from those in other NHPs. These data suggest that NHPs are mainly infected with G. duodenalis assemblage B and there might be geographical segregation and host-adaptation in assemblage B in NHPs.

[Medroxyprogesterone acetate therapy against antiangiogenesis of transplanted ovarian cancer in nude mice].

OBJECTIVE: To investigate the effects of medroxyprogesterone acetate (MPA) on angiogenesis and growth of transplanted human ovarian cancer cells in nude mice. METHODS: Ovarian cancer cell line COCI derived from human ovarian serous adenocarcinoma was transplanted into 30 nude mice, which were then randomized equally into 3 groups consisting of two treatment groups (in which MPA was administered at 60 and 120 mg/kg, respectively, twice a week for 4 weeks) and a control group. Six weeks later, the body mass of the nude mice was recorded and the morphology of tumor cells observed by electron microscope. The microvascular density (MVD) was examined by immunohistochemical staining with anti-human factor VIII antibody. RESULTS: Compared with the control group, the growth inhibitory rate in the two treatment groups were 23.76% and 43.80%, respectively, corresponding to the doses of 60 and 120 mg/kg. MVD of 60 mg/kg MPA group (3.64+/-0.02) and 120 mg/kg MPA group (2.11+/-0.12) was lower than that of the control group (5.14+/-0.74) (P<0.05 and P<0.01, respectively), and there was also significant difference between the two treatment groups (P<0.01). The morphological changes including compaction and margination of the nuclear chromatin, apoptotic bodies, and cell necrosis were significantly increased in the two treatment groups. CONCLUSIONS: MPA can inhibit the angiogenesis and growth of transplanted human ovarian cancer cells in nude mice in a dose-dependent manner, and its anticancer effect may involve induction of cell apoptosis as the result of its effect against angiogenesis.

Establishment of real time allele specific locked nucleic acid quantitative PCR for detection of HBV YIDD (ATT) mutation and evaluation of its application.

BACKGROUND: Long-term use of nucleos(t)ide analogues can increase risk of HBV drug-resistance mutations. The rtM204I (ATT coding for isoleucine) is one of the most important resistance mutation sites. Establishing a simple, rapid, reliable and highly sensitive assay to detect the resistant mutants as early as possible is of great clinical significance. METHODS: Recombinant plasmids for HBV YMDD (tyrosine-methionine-aspartate-aspartate) and YIDD (tyrosine-isoleucine-aspartate-aspartate) were constructed by TA cloning. Real time allele specific locked nucleic acid quantitative PCR (RT-AS-LNA-qPCR) with SYBR Green I was established by LNA-modified primers and evaluated with standard recombinant plasmids, clinical templates (the clinical wild type and mutant HBV DNA mixture) and 102 serum samples from nucleos(t)ide analogues-experienced patients. The serum samples from a chronic hepatitis B (CHB) patient firstly received LMV mono therapy and then switched to LMV + ADV combined therapy were also dynamically analyzed for 10 times. RESULTS: The linear range of the assay was between 1×10(9) copies/µl and 1 × 10(2) copies/µl. The low detection limit was 1 × 10(1) copies/µl. Sensitivity of the assay were 10(-6), 10(-4) and 10(-2) in the wild-type background of 1 × 10(9) copies/µl, 1 × 10(7) copies/µl and 1 × 10(5) copies/µl, respectively. The sensitivity of the assay in detection of clinical samples was 0.03%. The complete coincidence rate between RT-AS-LNA-qPCR and direct sequencing was 91.2% (93/102), partial coincidence rate was 8.8% (9/102), and no complete discordance was observed. The two assays showed a high concordance (Kappa = 0.676, P = 0.000). Minor variants can be detected 18 weeks earlier than the rebound of HBV DNA load and alanine aminotransferase level. CONCLUSIONS: A rapid, cost-effective, high sensitive, specific and reliable method of RT-AS-LNA-qPCR with SYBR Green I for early and absolute quantification of HBV YIDD (ATT coding for isoleucine) variants was established, which can provide valuable information for clinical antiretroviral regimens.