[Neurocognitive impairment and characteristics of neurocognitive performance among people with HIV on antiretroviral treatment].

Objective: Using two measuring tools to examine the prevalence and correlates of neurocognitive impairment (NCI) as well as characteristics of neurocognitive performance among people with HIV (PWH) on antiretroviral treatment (ART). Methods: A total of 2 250 treated PWH from the Comparative HIV and Aging Research in Taizhou (CHART) were recruited in Taizhou, Zhejiang province. The Chinese version of the Mini-mental State Examination (MMSE) and the International HIV Dementia Scale (IHDS) were used to evaluate their neurocognitive performance. Cluster analysis was conducted on the seven cognitive domains in the scale. Results: Among 2 250 treated PWH, 48.0% (1 080/2 250) were aged 45 to 89, 79.2% (1 782/2 250) were male, and 37.8% (852/2 250) had primary school education or below. The prevalence of neurocognitive impairment judged by MMSE and IHDS among HIV-infected people was 14.3% (321/2 250) and 31.8% (716/2 250), respectively. Aged 60 to 89 (aOR=2.63, 95%CI:1.52-4.56), depressive symptoms (aOR=5.58, 95%CI:4.20-7.40) and treatment with EFV (aOR=2.86, 95%CI:1.89-4.34) were main risk factors of NCI diagnosed by MMSE. Male (aOR=0.71, 95%CI:0.51-1.00), overweight (aOR=0.63, 95%CI:0.44-0.89), and high education level (aOR=0.11, 95%CI:0.05-0.25) were protective factors of NCI diagnosed by MMSE. Aged 60 to 89 (aOR=3.10, 95%CI:2.09-4.59), depressive symptoms (aOR=1.78, 95%CI:1.44-2.20) and treatment with EFV (aOR=1.79, 95%CI:1.41-2.29) were risk factors of NCI diagnosed by IHDS. Male (aOR=0.75, 95%CI:0.58-0.97), underweight (aOR=0.67, 95%CI:0.47-0.96), baseline CD4+ T lymphocyte (CD4) counts ≥350 cells/µl (aOR=0.69, 95%CI:0.53-0.91) and high education level (aOR=0.23, 95%CI:0.14-0.39) were protective factors of NCI diagnosed by IHDS. The neurocognitive performance of HIV-infected people can be divided into four main types. Among four types, age, gender, education level, alcohol drinking, depressive symptoms, waist-to-hip ratio, hypertension, diabetes, baseline CD4 counts and treatment with EFV were different statistically (all P<0.05). Conclusions: There are four main types of neurocognitive performance in treated PWH. The prevalence of NCI is high among this population, underscoring the need for tailored prevention and intervention.

[Application of anti-HCV and HCV RNA detection in intravenous drug users].

Objective: To explore the diagnostic value of anti-HCV and HCV RNA so as to provide an accurate and efficient detection strategy for the diagnosis of HCV in intravenous drug users. Methods: 527 plasma samples from intravenous drug users were collected, and preliminary anti-HCV ELISA screening test was performed. A recombinant immunoblot assay (RIBA) was used as confirmatory assay for reactive antibody samples. All samples were tested for HCV RNA, followed by analysis of anti-HCV screening test, RIBA and HCV nucleic acid test results. Results: Anti-HCV ELISA results were reactive in 386 out of 527 intravenous drug users and non-reactive in 141. Among the 386 reactive antibody samples detected by RIBA, 370 cases were anti-HCV positive, 6 cases were anti-HCV indeterminate and 10 cases were anti-HCV negative. Anti-HCV ELISA and RIBA positive coincidence detection rate was 95.85% (370/386), and 70.21% (370/527) among intravenous drug users. HCV RNA was negative in all 10 anti-HCV RIBA non-reactive samples. 376 anti-HCV RIBA-positive and indeterminate samples were tested for HCV RNA, of which 56.93% (300/527) were current HCV infection, and 14.42% (76/527) were past HCV infection. Among 141 anti-HCV ELISA negative samples, the residual risk by anti-HCV ELISA screening for HCV RNA was 1.52% (8/527). HCV viral load distribution among intravenous drug users showed that the high viral load value (>10(7) IU/ml) and low viral load values (< 10(2) IU/ml) accounted for 1.95% and 2.27%, respectively, while the samples with viral load value of 1×10(2) ~ 1×10(7) IU/ mL accounted for 95.78% (295/308), and were mainly distributed in 1×10(5) ~ 1×10(6) IU/ml (37.99%). ELISA + RIBA + NAT assay detection strategies had differentiated 300 cases of current HCV infection, 76 cases of past HCV infection and 10 cases of false positive anti-HCV results, while ELISA+NAT assay detection strategies had only detected 300 cases of current HCV infection. However, of the 386 positive subjects screened for antibodies, 10 (2.59%) were undifferentiated false positives. Conclusion: Intravenous drug users are the high-risk population of HCV infection with high prevalence and high viral load. Anti-HCV screening for intravenous drug users will have a certain degree of residual risk. Therefore, anti-HCV ELISA screening and nucleic acid detection strategy can accurately diagnose the current infected patients; however, it cannot distinguish the false positive results of antibody screening.

Long-term decomposed straw return positively affects the soil microbial community.

AIMS: In order to understand the response of soil microbial communities to the long-term of decomposed straw return, the modifications of soil microbial community structure and composition induced by more than 10 years of fresh and decomposed straw return was investigated and the key environmental factors were analysed. METHODS AND RESULTS: Phospholipid fatty acid analysis and high-through sequencing technique were applied to analyse the structure and composition of the soil microbial communities. Compared with fresh straw, returning decomposed straw increased the relative abundance of bacteria and fungi by 1·9 and 7·7% at a rate of ~3750 kg ha-1 , and increased by 23·1 and 5·7%, at a rate of ~7500 kg ha-1 respectively. The relative abundance of the bacteria related to soil nitrification increased, but the ones related to soil denitrification decreased with decomposed straw return, which led to higher total nitrogen contents in soils. Moreover, returning decomposed straw reduced pathogenic fungal populations (genus of Alternara), which had significantly positive correlation with soil electric conductivity. It indicated that the long-term of decomposed straw return might have lower risk of soil-borne disease mainly for the reasonable soil salinity. CONCLUSIONS: Long-term of decomposed straw return could provide suitable nutrient and salinity for healthier development of soil microbial community, both in abundance and structure, compared with fresh straw return. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of the study helps to better understand how the microbial community modifications induced by decomposed straw return benefit on soil health. The obtained key factors impacting soil microbial community variations is meaningful in soil health management under conditions of straw return.

[Clinical analysis of 36 cases of idiopathic intracranial hypertension complicated with iron deficiency anemia].

Objective: To investigate the clinical features, imaging findings and prognosis of idiopathic intracranial hypertension (IIH) patients complicated with iron deficiency anemia (IDA). Methods: A total of 307 cases of IIH patients hospitalized in Beijing Tongren Hospital were retrospectively screened between January 1, 2011 and February 28, 2018. There were 49 anemia cases (15.96%) and 45 IDA cases (14.66%), respectively. Finally, 36 IDA patients were enrolled. The clinical characteristics, imaging findings, treatment and prognosis of these patients were analyzed. Results: IIH combined with IDA was more common in women of childbearing age (34/36). There were 30 obese and overweight cases (83.33%), with multiple subacute or chronic course of disease. The visual symptoms in the early IIH patients were first diagnosed in the Department of Ophthalmology. The first symptom was headache with/without visual symptoms (27 cases (75%)). Head MRI detected empty sella or partial empty sella, and 2 cases of venous sinus thrombosis were found in DSA examination. Of the 34 female patients, 24 had simple menometrorrhagia or menstrual disorder. All patients were given methyl acetate to reduce the intracranial pressure and iron therapy. Five patients received low molecular weight heparin-warfarin sequential treatment, 5 cases underwent gynecologic surgery and 2 male cases received hemorrhoid operation. There were 7 cases underwent lumbar cisterna-peritoneal shunt for visual impairment. During the follow-up, intracranial pressure decreased and visual function of patients improved significantly. Conclusions: IIH is frequently found in obese or overweight women at childbearing age and IDA may be an important cause of IIH. IIH can cause serious irreversible visual impairment. Therefore, early identification and active treatment should be performed. Correction of anemia can significantly improve the clinical symptoms of IIH. Operation should be employed for IIH patients with poor visual function or rapid progress, in order to reduce intracranial pressure and improve prognosis as soon as possible.

Abnormal methylation of spermatozoa induced by benzo(a)pyrene in rats.

Epigenetic mutations caused by pollutants are possibly linked to many diseases. Benzo(a)pyrene (BaP) is one of the most representative air pollutants and has aroused wide concern because of its strong carcinogenicity. The reproductive toxicity induced by BaP has been identified, but little is known about the characteristics of the methylation changes induced by BaP. In this study, a methylated DNA immunoprecipitation sequencing method was used to detect the methylation of sperm DNA of rats exposed to BaP. Compared with the respective genes in normal rats, there were 3227 hypomethylated genes and 828 hypermethylated genes after BaP exposure. Gene ontology enrichment analysis reported that differentially methylated genes (DMGs) were enriched in the localization, single-multicellular organism process and plasma membrane. Kyoto Encyclopedia of Genes and Genomes pathway analysis showed that the DMGs were significantly enriched in the Ras signalling pathway, Rap1 signalling pathway, pancreatic secretion and neuroactive ligand-receptor interaction. DisGeNET disease spectrum analysis showed that DMGs were associated with infertility and certain genetic diseases. Further research needs to be done to explore whether these abnormal methylation are transgenerational.

[Assessing right ventricular function in patients with pulmonary artery hypertension based on noninvasive measurements: correlation between cardiac MRI, ultrasonic cardiogram, multidetector CT and right heart catheterization].

Objective: To compare the value of cardiac MRI (CMRI), ultrasonic cardiogram (UCG), multidetector CT (MDCT) in assessing right ventricular function (RV) in patients with PAH. Methods: A total of 31 consecutive patients with PAH (17 males and 14 females, 55±12 years) in Beijing Chao-Yang Hospital from August 2012 to February 2014 were prospectively enrolled. All patients underwent CMRI to get parameters including right ventricular end-systolic volume (ESV), end-diastolic dimension (EDV), stroke volume (SV), ejection fraction (EF), ventricular mass index (VMI). UCG parameters included Tei index, RV fractional area change (FAC), ESV, EDV. MDCT parameters included right /left ventricular internal diameter (RVd/LVd), right /left ventricular diastole maximum area (RVa/LVa), Cobb angle.These parameters obtained by MRI, UCG and MDCT were correlated with those of RHC respectively by Spearman or Pearson correlation analysis. Results: Six minutes walk distance had moderate negative correlation with CMRI-EF (40±9), VMI 44-115(71±20) g/m(2,) Cobb angle(67°±12°); RHC-SV had moderate negative correlation with CMRI-SV(57±21) ml, EF, VMI, UCG-EF(41±14), Tei(0.82±0.29), FAC(30±9), RVd(45±7) mm, RVa(2 484±596) mm(2), Cobb angle ; Right cardiac work index had moderate negative correlation with CMRI-EF, RVd and Cobb angle. Conclusions: MRI-EF is the best parameter to reflect RV function. CMRI is the optimal method to assess RV function, and then is the MDCT and the last is UCG.

Nrf2-ARE signaling pathway regulates the expressions of A1R and ENT1 in the brain of epileptic rats.

OBJECTIVE: To explore the behavioral changes and the expressions of the A1 receptor (A1R) and balanced nucleoside transporter-1 (ENT1) in the brain of epileptic rats after activating the NF-E2-related factor 2 (Nrf2)-ARE signaling pathway. MATERIALS AND METHODS: Adult male Sprague-Dawley (SD) rats were randomly divided into normal control group, epilepsy group, and t-butylhydroquinone (tBHQ) group, with 10 rats in each group. Lithium-pilocarpine induced epilepsy model in rats was established. The first epileptic latency and seizure frequency within 1 hour were observed in each group using the Racine scoring system. HE (Hematoxylin and Eosin) staining was used to observe the pathological lesions in the brain tissue of each group. The expressions of A1R, ENT1, and relative genes in Nrf2-ARE pathway in rat hippocampus was detected by immunohistochemistry and Western blot. RESULTS: Compared with rats in epileptic group, the first seizure latency was prolonged and the seizure frequency decreased in tBHQ group (p<0.05). The degree of brain lesions in tHBQ group was lighter than that of epilepsy group. ENT1 expression in rat hippocampus of epileptic group was significantly upregulated than that of normal control group and tBHQ group. Besides, the protein levels of A1R, Nrf2, HO-1, and ARE in rat hippocampus of epilepsy group markedly decreased compared with those of normal control group. However, protein expressions of A1R, Nrf2, HO-1, and ARE proteins in rat hippocampus of tBHQ group were markedly upregulated. CONCLUSIONS: Activation of the Nrf2-ARE signaling pathway can reduce the pathological damage of rat hippocampal neurons, prolong the latency of seizures, and reduce the degree of epileptic seizure in rats.

Fasudil alleviates hepatic fibrosis in type 1 diabetic rats: involvement of the inflammation and RhoA/ROCK pathway.

OBJECTIVE: Rho-associated kinases (ROCKs) are recognized to be involved in many pathophysiological processes caused by hyperglycemia. We performed experiments to evaluate the effects of fasudil, the Rho/ROCK inhibitor, on preventing hepatic fibrosis in type 1 diabetic rats and to elucidate the underlying mechanisms. MATERIALS AND METHODS: Sprague-Dawley (SD) rats were randomly divided into five groups: normal control (NC), untreated diabetic (DM), low-dose fasudil-treated (L-Fas), high-dose fasudil-treated (H-Fas) and captopril-treated (Cap) groups. Streptozotocin was injected to establish the diabetes model. Alanine aminotransferase (ALT), aspartate aminotransferase (AST) and inflammatory factors such as tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), were analyzed. Hematoxylin and eosin (HE) and Masson's trichrome staining were used for histological observations. The expression of transforming growth factor-ß (TGF-ß1), metalloproteinase-9 (MMP-9)/tissue inhibitor of metalloproteinase-1 (TIMP-1), collagen type Iα (Coll α1), nuclear factor-kappa B (NF-κB) and ROCK-1 were measured to investigate the mechanisms involved in fibrosis. RESULTS: The DM group exhibited hepatic fibrosis with remarkable liver damage and inflammation reaction by the activation of the NF-κB pathway. Treatment with fasudil or captopril suppressed not only the inflammation reaction but also the accumulation of the extracellular matrix due to the downregulation of TGF-ß1 and MMP-9/TIMP-1, which induces the amelioration of the liver fibrosis with diabetes. Furthermore, fasudil significantly attenuated the activation of ROCK-1 and NF-κB in the livers of diabetic rats. CONCLUSIONS: These results suggest that fasudil exert anti-inflammation actions and markedly decrease the accumulation of extracellular matrix. Fasudil is a good candidate agent for treating hepatic fibrosis in diabetes.

Single-Atom Demonstration of the Quantum Landauer Principle.

One of the outstanding challenges to information processing is the eloquent suppression of energy consumption in the execution of logic operations. The Landauer principle sets an energy constraint in deletion of a classical bit of information. Although some attempts have been made to experimentally approach the fundamental limit restricted by this principle, exploring the Landauer principle in a purely quantum mechanical fashion is still an open question. Employing a trapped ultracold ion, we experimentally demonstrate a quantum version of the Landauer principle, i.e., an equality associated with the energy cost of information erasure in conjunction with the entropy change of the associated quantized environment. Our experimental investigation substantiates an intimate link between information thermodynamics and quantum candidate systems for information processing.

Experimental Verification of a Jarzynski-Related Information-Theoretic Equality by a Single Trapped Ion.

Most nonequilibrium processes in thermodynamics are quantified only by inequalities; however, the Jarzynski relation presents a remarkably simple and general equality relating nonequilibrium quantities with the equilibrium free energy, and this equality holds in both the classical and quantum regimes. We report a single-spin test and confirmation of the Jarzynski relation in the quantum regime using a single ultracold ^{40}Ca^{+} ion trapped in a harmonic potential, based on a general information-theoretic equality for a temporal evolution of the system sandwiched between two projective measurements. By considering both initially pure and mixed states, respectively, we verify, in an exact and fundamental fashion, the nonequilibrium quantum thermodynamics relevant to the mutual information and Jarzynski equality.

Measurement of the absolute velocity of blood flow in early-stage chick embryos using spectral domain optical coherence tomography.

Doppler optical coherence tomography (OCT) is a noninvasive imaging modality that provides quantitative flow information with high spatial and temporal resolution. However, it is only sensitive to the flow velocity vector parallel to the incident beam. To calculate the absolute velocity, it is necessary to obtain the angle between the incident beam and flow field. In this paper, we describe a practical method to measure the Doppler angle based on the structural information of blood vessels extracted from spectral domain OCT images. In this method, a normal sectional scan of the vessel is performed where the probe beam is perpendicular to the vessel. Next, the axial diameter (Z direction) of the vessel (DA) was measured in the acquired image. For a certain scan in which the probe beam is oblique to the blood vessel, the axial diameter of the vessel (DA') can be measured. Thus, the Doppler angle can be calculated depending on the ratio of DA and DA', and absolute blood flow was determined. We validate this method in a capillary tube as well as in large blood vessels of early-stage chick embryos. This technique is suitable for early-stage embryo blood-flow measurement because most of the blood vessels are easily differentiated from the transparent surrounding structures during that time.

[Effect of probucol on preventing contrast-induced nephropathy in patients undergoing percutaneous coronary intervention].

Objective: To investigate the preventive effect, possible mechanism and safety of probucol on contrast-induced nephropathy (CIN) after percutaneous coronary intervention (PCI) in patients with coronary heart disease (CHD). Methods: A total of 641 patients with coronary heart disease were consecutively enrolled from Department of Cardiology, in Tianjin Chest Hospital, Tianjin TEDA International Cardiovascular Hospital, Tianjin First Central Hospital, Tianjin Fourth Central Hospital. They were randomly divided into probucol group (n=321) and control group (n=320). The probucol group was given oral probucol 500 mg twice daily for day 0 to day 3 after PCI; the control group was given only conventional therapy. All patients were given intravenous drip 0.9% sodium chloride solution before 12 to 24 hours of operation. The levels of serum creatinine (Scr), blood urea nitrogen (BUN), evaluate glomerular filtration rate (eGFR), cystatin C (Cys-C), and high-sensitivity C-reactive protein (hs-CRP), neutrophil gelatinase associated lipocalin (NGAL), superoxide dismutase (SOD) and glutathione (GSH) were measured before and 72 h after the PCI operation in both groups. The incidence rates of CIN, the adverse events during hospitalization and postoperative 14-day follow-up were recorded in two groups. Results: There was no statistically significantly difference in the levels of Scr, BUN, eGFR, Cys-C, hs-CRP, NGAL, SOD and GSH between the two groups before PCI (P>0.05). The levels of serum Scr, BUN, Cys-C, hs-CRP, NGAL, SOD and GSH after operation in the two groups were higher than those before the operation (P<0.05). The levels of hs-CRP and NGAL in the probucol group were lower than those in the control group [(10±4) vs (11±4)mg/L, (25±8)vs (34±7)U/ml, P<0.05]. The levels of eGFR, SOD and GSH in probucol group were higher than those in control group [(80±27) vs (72±26) ml·min(-1)·1.73 m(-2,) (67±9) vs (58±8)U/ml, (4.6±0.9) vs (3.9±0.8)U/ml, P<0.05]. The incidence of CIN was 4.0% in the probucol group and 10.9% in the control group, and the difference was statistically significant (P<0.05, χ(2)=-3.31). Multivariate Logistic regression analysis showed that probucol was an independent protective factor for CIN (OR=0.334, 95%CI 0.172-0.648, P=0.001). There were no adverse events such as myasthenia gravis, abnormal liver function and cardiovascular events during the hospitalization and 14-day follow-up. Conclusions: Probucol can reduce the incidence of contrast-induced nephropathy after PCI. The protection mechanism is related with its anti-inflammatory and anti-oxidative stress effects, and it has good safety.

[Oncolytic property of HSV-1 recombinant viruses carrying the human IL-12].

Objective: Constructed the recombinant HSV-1 deleted ICP47 and inserted human IL-12, and investigate the virus' replication ability and oncolytic property in vitro and vivo. Methods: The recombinant HSV-1 deleting ICP47 (MH1005) and then inserting human IL-12 (MH1006) were obtained with bacterial artificial chromosome technology.The replication ability and the efficiency of inhibiting tumor were detected in several nerve tumor cell lines infected with HSV-wt, MH1005 and MH1006 respectively.The murine tumor model was established by subcutaneous inoculation Neuro-2a cells on both sides of mice back respectively.A dosage of 2×10(6) PFU of HSV-wt, MH1001(recombinant HSV-1 deleted IR), MH1005, MH1006 and Mock were injected 3 times intratumorally on one side of mice back in every 3 days, the tumor volume and survival rate of the mice were measured. Results: The replication abilities of MH1005, MH1006 and HSV-wt in 293FT cells were insignificant (P>0.05); the replication abilities of recombinant HSV-1 in G422 and Neuro-2a were higher than that in SK-N-SH; and the nerve tumor cells could be inhibited significantly by recombinant HSV-1.After 15 days of treatment, on the mouse backside with injection treatment, the tumor volumes of group HSV-wt (6 267±484), MH1001 (5 730±1 071), MH1005 (4 537±538)and MH1006 (4 150±476)mm(3) were smaller than that of group Mock (6 957±722) mm(3) significantly (all P<0.01); on the mouse backside without injection treatment, the tumor volumes of group MH1005 (5 952±607) and MH1006 (5 473±661) mm(3) were smaller than those of HSV-wt (6 785±1 063), MH1001 (6 774±808) and Mock (6 957±190) mm(3) significantly (all P<0.05); after 35 days of treatment, the mice survival rates of group MH1005 (100%) and MH1006 (100%) were higher than those of MH1001 (67%), HSV-wt (50%) and Mock (33%) significantly (all P<0.05). Conclusion: MH1005 and MH1006 can infect nerve tumor cells and replicate at high level, the viruses not only kill tumor cells directly but also induce immunological rejection to tumor, and prolong the survival of mice bearing tumor.

Characterizing and marker-assisting a novel chili pepper (Capsicum annuum L.) yellow bud mutant with cytoplasmic male sterility.

Cytoplasmic male sterility (CMS) in pepper is a better way to produce hybrid seeds compared to manual production. We used the two sequence characterized amplified region (SCAR) markers (CRF-SCAR and CMS-SCAR130) in CMS pepper, to identify the genotype. We assembled two CMS yellow bud mutants (YBM; YBM12-A and YBM12-B). This mutation in leaf color is controlled by a single dominant nuclear gene. The aim was to create a new hybrid seed production method that reduces the costs and increases F1 hybrid seed purity. The results suggest that the CRF-SCAR and CMS-SCAR130 markers can be used together in multiple generations to screen for restorer or maintainer genes. We found the marker linked to the restorer gene (Rf) in the C-line and F1 hybrids, as well as partially in the F2 generation, whereas it was not found in the sterile YBM12-A or the maintainer line YBM12-B. In the F2 population, sterility and fertility segregated at a 3:1 ratio based on the CRF-SCAR marker. A 130 bp fragment was produced in the YBM12-A, F1, and F2 populations, suggesting that these lines contained sterile cytoplasm. A 140 bp fragment present in the YBM12-B and C-line indicated that these lines contained normal cytoplasm. In addition, we identified some morphological characters distinguishing sterile and fertile buds and flowers that may be linked to the sterility gene. If more restorer lines are identified, CMS expressing the YBM trait can be used in hybrid seed production.

Defective expression of polarity protein Par3 promotes cervical tumorigenesis and metastasis.

OBJECTIVES: The aim of this study was to investigate whether the partition-defective 3 protein (Par3) regulates cervical carcinoma growth and metastasis. MATERIALS AND METHODS: Immunohistochemistry (IHC) was used to analyze the expression of Par3 protein in samples from 89 cervical squamous cell carcinoma (CSCC) patients among Uyghur women. The specific short hairpin (shRNA) vector as well as eu- karyotic expression vector of PARD3 was transfected into SiHa cell lines. The variation of migration and invasion after transfection was determined using Transwell assays, cell cycle, and apoptosis were assayed by flow cytometry, respectively. RESULTS: The incidence of CSCC was associated with reduced expression of Par3. Downregulation of Par3 was significantly associated with more advanced tumors (i.e., higher histological grade, lymph node involvement, and higher tumor stages) (p < 0.05 for all). Lost expression of Par3 promotes prolif- eration, inhibits apoptosis, and enhances migration and invasion. Loss of Par3 induces MMP9 expression and epithelial to mesenchymal transition (EMT) related genes (N-cadherin, E-cadherin, and ß-catenin) expression changed in SiHa cells. CONCLUSIONS: The reduced Par3 expression in cervical cancer indicates tumor-suppressive properties of Par3 that may be a marker of poor prognosis in cervical cancer patients, and the molecular determinants of epithelial polarity which have tumorigenesis enhancing impact, might through EMT.

Evaluation of a loop-mediated isothermal amplification assay based on hrpZ gene for rapid detection and identification of Pseudomonas syringae pv. lachrymans in cucumber leaves.

AIMS: Cucumber angular leaf spot caused by Pseudomonas syringae pv. lachrymans (Psl) is an important and destructive disease worldwide, and no effective technique has been developed for the control of the pathogen. Detection of infection or latent in cucumber plants is critical to evaluate disease progress and strengthening management to avoid a serious epidemic in the fields. In this paper, we developed a rapid and sensitive method for detection of Psl using an isothermal method known as loop-mediated amplification (LAMP). METHODS AND RESULTS: A set of six primers was designed to amplify the gene coding for the hrpZ, and conditions for detection were optimized to complete in 60 min at 67°C, and the amplification were confirmed through gel electrophoresis or visually inspected using calcein stain. The specificity of LAMP primers set was widely validated on Psl and nontarget strains. In sensitivity testing, LAMP allowed detection as low as 104  CFU per ml bacterial cells without DNA extraction. The novel method was also applied for detecting Psl in infected cucumber leaves, and even the early onset of disease can be detected by the assay. CONCLUSIONS: This study confirmed that the novel developed LAMP assay is an easy, rapid and sensitive method for the detection of Psl in infected leaves. SIGNIFICANCE AND IMPACT OF THE STUDY: The method is suitable for direct detection of Psl without strain enrichment and complex DNA extraction from samples in the field, and hence it has the capability to be used for on-site disease diagnosis and field surveys.

[The analysis of the clinical records diagnosed as Takayasu's arteritis with pulmonary vascular involvement].

OBJECTIVE: To enhance the understanding of Takayasu's arteritis with pulmonary vascular involvement through the analyses on clinical features, imaging findings, misdiagnoses and treatments. METHODS: A retrospective study was conducted to analyze the clinical records of patients diagnosed as Takayasu's arteritis with pulmonary vascular involvement admitted to Beijing Chaoyang Hospital from January 2004 to December 2014. RESULTS: In recent 10 years, there were 90 patients diagnosed as Takayasu's arteritis in Beijing Chaoyang Hospital, 33 of them were involved with pulmonary arterial, which account for 36.7%, while 12 cases were involved with pulmonary arteries alone, which account for 13.3% of all. Among the 33 patients, dyspnea (23 cases, 69.7%) was the most common symptom, followed by hemoptysis (19 cases, 57.6%) and Vascular murmur (23 cases, 69.7%), pleural effusion (8 cases, 24.2%), unequal blood pressure of upper limbs (4 cases, 12.1%) were the main signs. The diagnostic time varied significantly, from 1 month to 10 years. 21 patients were misdiagnosed and the misdiagnosis rate was 63.6%. CONCLUSION: Takayasu's arteritis with pulmonary vascular involvement is not rare. The clinical manifestation of Takayasu's arteritis is unspecific and misdiagnosis rate is relatively high. Therefore, we should raise awareness of Takayasu's arteritis and detailed clinical examination will be helpful to reduce the misdiagnosis rate.

Autologous peripheral blood stem cell transplantation to treat thromboangiitis obliterans: preliminary results.

OBJECTIVE: To investigate the efficiency of autologous peripheral blood stem cell transplantation (APBSCT) to treat severe the lower limbs ischemia caused by thromboangiitis obliterans (TAO). PATIENTS AND METHODS: From April 2007 to December 2014, a total of 64 patients with TAO (80 affected limbs) received APBSCT at our hospital. The treatment effect was evaluated by subjective indicators including pains and cold sensation of the affected limbs, combined with objective indicators including claudication distance, ankle brachial index (ABI), transcutaneous oxygen pressure (TcPO2) and skin temperature. RESULTS: Five patients (with 5 affected limbs) suffered from necrosis below the middle of the leg 4 weeks after transplantation and received amputation. For the remaining 59 patients (75 affected limbs), pain and cold sensation of the affected limbs were improved with varying extent 3 months after transplantation; there were statistically significant differences in pain score and cold sensation score of the affected limbs before and after APBSCT (p<0.05). Claudication distance, ABI, TcPO2 and skin temperature were also improved. Claudication distance increased from 85.69 m ± 43.48 m to 36.5 ± 9.88 mmHg, and the skin temperature of the lower limbs increased from 27.70 °C ± 0.53 °C to 33.49 °C ± 0.60 °C. All four indicators were considerably improved after APBSCT (p<0.05). Arteriography was performed for 75 affected limbs in 59 patients 6 months after transplantation and found that new collateral vessels were formed in the affected limbs. No patients were complicated by retinal hyperplasia, malignant tumors, myocardial infarction and cerebral infarction during the follow-up examinations; no patients underwent symptom aggravation during 9-48 month follow-up (average, 28.5 months). CONCLUSIONS: APBSCT is an easy, safe and reliable treatment for ischemia of lower limbs, especially for those with poor distal arterial outflow tract in the lower limbs that do not permit bridging.

Multiple displacement amplification of whole genomic DNA from urediospores of Puccinia striiformis f. sp. tritici.

Biotrophic fungi, such as Puccinia striiformis f. sp. tritici, because they cannot be cultured on nutrient media, to obtain adequate quantity of DNA for molecular genetic analysis, are usually propagated on living hosts, wheat plants in case of P. striiformis f. sp. tritici. The propagation process is time-, space- and labor-consuming and has been a bottleneck to molecular genetic analysis of this pathogen. In this study we evaluated multiple displacement amplification (MDA) of pathogen genomic DNA from urediospores as an alternative approach to traditional propagation of urediospores followed by DNA extraction. The quantities of pathogen genomic DNA in the products were further determined via real-time PCR with a pair of primers specific for the ß-tubulin gene of P. striiformis f. sp. tritici. The amplified fragment length polymorphism (AFLP) fingerprints were also compared between the DNA products. The results demonstrated that adequate genomic DNA at fragment size larger than 23 Kb could be amplified from 20 to 30 urediospores via MDA method. The real-time PCR results suggested that although fresh urediospores collected from diseased leaves were the best, spores picked from diseased leaves stored for a prolonged period could also be used for amplification. AFLP fingerprints exhibited no significant differences between amplified DNA and DNA extracted with CTAB method, suggesting amplified DNA can represent the pathogen's genomic DNA very well. Therefore, MDA could be used to obtain genomic DNA from small precious samples (dozens of spores) for molecular genetic analysis of wheat stripe rust pathogen, and other fungi that are difficult to propagate.

Temporal dynamics and population genetic structure of Fusarium graminearum in the upper Midwestern United States.

Fusarium graminearum sensu stricto causes Fusarium head blight (FHB) in wheat and barley, and contaminates grains with several trichothecene mycotoxins, causing destructive yield losses and economic impact in the United States. Recently, a F. graminearum strain collected from Minnesota (MN) was determined to produce a novel trichothecene toxin, called NX-2. In order to determine the spatial and temporal dynamics of NX-2 producing strains in MN, North Dakota (ND) and South Dakota (SD), a total of 463 F. graminearum strains were collected from three sampling periods, 1999-2000, 2006-2007 and 2011-2013. A PCR-RFLP based diagnostic test was developed and validated for NX-2 producing strains based on polymorphisms in the Tri1 gene. Trichothecene biosynthesis gene (Tri gene)-based polymerase chain reaction (PCR) assays and ten PCR-restriction fragment length polymorphism (RFLP) markers were used to genotype all strains. NX-2 strains were detected in each sampling period but with a very low overall frequency (2.8%) and were mainly collected near the borders of MN, ND and SD. Strains with the 3ADON chemotype were relatively infrequent in 1999-2000 (4.5%) but increased to 29.4% in 2006-2007 and 17.2% in 2011-2013. The distribution of 3ADON producing strains also expanded from a few border counties between ND and MN in 1999-2000, southward toward the border between SD and MN in 2006-2007 and westward in 2011-2013. Genetic differentiation between 2006-2007 and 2011-2013 populations (3%) was much lower than that between 1999-2000 and 2006-2007 (22%) or 1999-2000 and 2011-2013 (20%) suggesting that most change to population genetic structure of F. graminearum occurred between 1999-2000 and 2006-2007. This change was associated with the emergence of a new population consisting largely of individuals with a 3ADON chemotype. A Bayesian clustering analysis suggested that NX-2 chemotype strains are part of a previously described Upper Midwestern population. However, these analyses also suggest that the NX-2 isolates could represent a distinct population, but that interpretations of population assignment are influenced by the small number of NX-2 strains available for analysis.