Occurrence and Genetic Diversity of Grapevine berry inner necrosis virus from Grapevines in China.

To investigate the prevalence and genetic diversity of Grapevine berry inner necrosis virus (GINV) in China, 195 grapevine samples from 15 Chinese provinces and regions were tested using reverse-transcription polymerase chain reaction. The samples included symptomatic and asymptomatic cultivars, with 35.9% (70 of 195) of samples testing positive for GINV. Seventeen samples had obvious ring spot symptoms, and 94.1% (16 of 17) tested positive for GINV, suggesting that GINV may be highly associated with the ring spot symptom. The genetic diversity of GINV isolates was analyzed based on the partial nucleotide and amino acid sequences of the coat protein (CP) and movement protein (MP) genes. Phylogenetic analyses of the MP and CP gene sequences divided the GINV isolates into three groups. The majority of the Chinese isolates were in groups 1 and 2, and only one Chinese isolate, along with a previously reported Japanese isolate, was in group 3. This is the first report on the genetic diversity of GINV isolates and their prevalence and distribution in China.

Development of an indirect ELISA with epitope on nonstructural protein of Muscovy duck parvovirus for differentiating between infected and vaccinated Muscovy ducks.

UNLABELLED: The aim of this study was to develop an indirect enzyme-linked immunosorbent assay (i-ELISA) based on epitope AA503-509 (RANEPKE), which is on nonstructural protein of Muscovy duck parvovirus (MDPV). Sera (100) from negative and vaccinated Muscovy ducks were compared with infected sera (240) to establish the cut-off value of this i-ELISA. There was a significant difference between the positive and negative populations (P < 0·05). The adoption of this positive-negative threshold value for this i-ELISA assay resulted in specificity of 98·0%. This i-ELISA could be used as a diagnostic tool for differentiating infected Muscovy ducks from Muscovy ducks vaccinated with inactivated virus. SIGNIFICANCE AND IMPACT OF THE STUDY: In this study, we developed an i-ELISA based on epitope AA503-509 (RANEPKE), which is on nonstructural protein of MDPV. This i-ELISA could be used as a diagnostic tool for differentiating infected Muscovy ducks from Muscovy ducks vaccinated with inactivated virus.

Detection and sequence analysis of grapevine virus B isolates from China.

The presence of grapevine virus B (GVB) was detected in 188 grapevine samples from China by double antibody sandwich ELISA (DAS-ELISA) and reverse transcription-PCR (RT-PCR). The accuracy of detection by RT-PCR was confirmed by sequencing amplified PCR fragments. Seventeen samples were GVB-positive by DAS-ELISA and five by RT-PCR. The isolate QMW proved to be positive by RT-PCR only, and four isolates (DGWH, DGW, QM, and JFL) could be detected by both methods. Among the five GVB-positive samples detected by RT-PCR, two isolates were originally collected from Henan province and three from Liaoning province. The expected 722 bp DNA fragment, covering partial ORF3 through partial ORF5, was amplified from the five GVB infected samples. Sequence analysis revealed that the molecular variants΄ composition of GVB in the different isolates was complex. Clones of DGWH, DGW, QM, and JFL isolate shared high nucleotide identities, while the identities among the clones of isolate QMW varied. The variants of GVB isolates obtained in this study showed nucleotide identities from 81.1% to 97.9% among themselves, and 79.1% to 98.5% identity with five previously published GVB isolates in NCBI. The alignment of partial ORF3 and the phylogenetic relationships of ORF4 revealed that the molecular variants of Chinese GVB isolates could be clustered into three groups. Only isolate DGW was in the same group with the reported GVB isolates from other countries; the other four GVB isolates in this study were clustered into two groups.

[A prospective cohort study on BMI levels and risk of acute pancreatitis].

Objective: To investigate the effects of body mass index (BMI) levels at different baseline on the risk of new-onset acute pancreatitis (AP). Methods: The subjects were from the Kailuan Study Cohort and divided into 3 groups according to baseline BMI levels: BMI<24 kg/m2, normal weight; BMI 24-28 kg/m2, overweight; BMI≥28 kg/m2, obesity. The incidence of new-onset AP in these three groups was analyzed. The survival curve was plotted by Kaplan-Meier method, the cumulative incidence was calculated and tested by log-rank method. Multivariate Cox proportional hazards regression model was used to calculate HR of baseline BMI levels for AP. Results: A total of 123 841 subjects were included and followed up for (11.94±2.13) years, during which, 395 cases were found with AP. The incidence of AP was 2.67 per 10 000 person years in total population, and the incidences of AP were 2.20, 2.72 and 3.58 per 10 000 person-years in the normal, overweight and obesity groups, respectively. The cumulative incidences of AP was 0.32%, 0.40% and 0.49% in normal, overweight and obesity groups, respectively, which showed a significant inter-group difference by log-rank test (χ 2=13.17,P<0.01). The results of multivariable adjusted Cox proportional hazards regression model analysis indicated that obesity group (HR=1.45, 95%CI: 1.10-1.92) had a higher risk for AP compared with the normal BMI group. The subgroup analyses by age and sex showed that compared with the normal weight group,the HRs for AP in the obesity group was 1.58(95%CI:1.14-2.19) and 1.40(95%CI:1.03-1.90) among subjects younger than 60 years old and male subjects, respectively. After excluded onset AP within two years from baseline,with a control group from normal weight,the results of multivariate Cox proportional hazards regression model analysis indicated that the AP in the obesity group was 1.60 (95%CI: 1.18-2.15). Conclusion: Obesity may increase the risk of developing AP, particularly among young and middle-aged men.

A major T cell antigen of Mycobacterium leprae is a 10-kD heat-shock cognate protein.

Several mycobacterial antigens, identified by monoclonal antibodies and patient sera, have been found to be homologous to stress or heat-shock proteins (hsp) defined in Escherichia coli and yeast. A major antigen recognized by most Mycobacterium leprae-reactive human T cell lines and cell wall-reactive T cell clones is a 10-kD protein that has now been cloned and sequenced. The predicted amino acid sequence of this protein is 44% homologous to the hsp 10 (GroES) of E. coli. The purified native and recombinant 10-kD protein was found to be a stronger stimulator of peripheral blood T cell proliferation than other native and recombinant M. leprae proteins tested. The degree of reactivity paralleled the response to intact M. leprae throughout the spectrum of leprosy. Limiting-dilution analysis of peripheral blood lymphocytes from a patient contact and a tuberculoid patient indicated that approximately one third of M. leprae-reactive T cell precursors responded to the 10-kD antigen. T cell lines derived from lepromin skin tests were strongly responsive to the 10-kD protein. T cell clones reactive to both the purified native and recombinant 10-kD antigens recognized M. leprae-specific epitopes as well as epitopes crossreactive with the cognate antigen of M. tuberculosis. Further, the purified hsp 10 elicited strong delayed-type hypersensitivity reactions in guinea pigs sensitized to M. leprae. The strong T cell responses against the M. leprae 10-kD protein suggest a role for this heat-shock cognate protein in the protective/resistant responses to infection.

First Report of Grapevine leafroll-associated virus 4 and 5 in Grapevines in China.

Grapevine leafroll disease (GLD) is one of the most important diseases of grapevines worldwide. Nine serologically distinct viruses in the Closteroviridae family are associated with GLD. Previous studies reported that Grapevine leafroll-associated virus (GLRaV) -1, -2, -3, and -7 were present in grapevines in China with GLRaV-1 and -3 the predominant viruses associated with GLD (1). To confirm if GLRaV-4 and -5 were also present in China, 36 dormant canes from individual vines of 29 cultivars that showed GLD leaf symptoms during the growing season were collected from the germplasm collection plot of the Research Institute of Pomology, Chinese Academy of Agricultural Sciences. Total RNA extracted by a silica capture protocol (2) from phloem-enriched bark of 36 samples was tested separately for GLRaV-4 and -5 by reverse transcription (RT)-PCR using virus-specific primers. Primers LR4F (5'-ACATTCTCCACCTTGTGCTTTT-3') and LR4R (5'-CATACAAGCGAGTGCAATTAC-3') (4) were used to amplify a 321-bp fragment corresponding to a partial region of the HSP70 gene from GLRaV-4. One sample from cv. Autumn Royal was infected by GLRaV-4. The amplicon was cloned and a single clone was sequenced (GenBank Accession No. GQ246624) that showed 99% nucleotide identity with a corresponding region of a GLRaV-4 isolate from the United States (Accession No. AF039553). Since antiserum specific to GLRaV-4 was unavailable, a second pair of primers, LR4CP-F (5'-GGTGTCCAGCGCTTCCAA-3') and LR4CP-R (5'-GCCAGAGAAGCATCGTAA-3'), was designed on the basis of the sequence of GLRaV-4 from Chile (Accession No. EU746620) that amplified a 300-bp fragment specific to the coat protein gene of GLRaV-4. The amplicon was cloned and a single sequence (Accession No. GQ479041) was compared with a corresponding nucleotide sequence of GLRaV-4 from Chile (Accession No. EU746621) showing 99% identity. A sample from cv. Malaga Rose was positive when tested by ELISA with antibodies specific to GLRaV-5 (Neogen Europe, Ltd. Scotland, UK) and this was confirmed by amplification of a 690-bp fragment corresponding to the GLRaV-5 coat protein gene using virus-specific primers LR5F (5'-CCCGTGATACAAGGTAGGACA-3') and LR5R (5'-CAGACTTCACCTCCTGTTAC-3') (3). The amplicon was cloned and a single clone was sequenced (Accession No. GQ246625) that showed 95% nucleotide identity with the CP gene sequence of GLRaV-5 from Argentina (Accession No. EU815935). To our knowledge, this is the first report of GLRaV-4 and -5 in grapevines in China. Confirmation of these viruses in China is very important for producing virus-free plants and this information also will be helpful in developing a multiplex RT-PCR assay to simultaneously detect multiple GLRaVs and helpful with studies on the molecular variability of these viruses. References: (1) Y. Dong et al. China Fruits 6:9, 2005. (2) X. Foissac et al. Acta Hortic. 550, 37, 2001. (3) X. Good and J. Monis. Phytopathology 91:274, 2001. (4) F. Osman et al. J. Virol. Methods 141:22, 2007.

An effective experimental method and apparatus for unsteady water vapor condensation investigation in high speed expansion flow.

This paper presents an effective experimental method and system for mechanism study of unsteady water vapor condensation encountered in high speed expansion flow. We proposed an experimental method and designed a simplified expansion system to fulfill the study. Tunable diode laser absorption spectroscopy (TDLAS) and light sheet technique are integrated in the system to monitor the unsteady condensation process in a high speed expansion flow generated by the expansion system. Two near infrared water vapor absorption transitions (1395.0 nm and 1409.27 nm) and one near infrared methane absorption transition (1653.73 nm) are applied in the TDLAS measurement to measure the transient flow parameters during the condensation process. Using the experimental method, time dependent condensation processes are monitored with different expansion time scales. The light sheet results visually reveal the condensation phenomena during the expansion process, while TDLAS results quantitatively follow the condensation process. The experimental results are compared with computational fluid dynamics simulations and a good agreement between them is observed, which indicates that the presented experimental method and system is effective in investigating unsteady water vapor condensation in high speed expansion flow.

Molecular cloning of a gene selectively induced by gamma interferon from human macrophage cell line U937.

A cDNA clone encoding a gamma interferon (IFN-gamma)-inducible mRNA in human cells of the macrophage lineage was isolated and characterized. The corresponding gene, gamma.1, was selectively induced by IFN-gamma, responding a hundredfold better to IFN-gamma than to IFN-alpha. The induction was rapid and transient, with maximal mRNA accumulation at about 3 h and decline to the basal level after 48 h. Transcriptional activation could be detected as early as 5 min after IFN-gamma stimulation and accounted entirely for the mRNA accumulation. The induction of gamma.1 by IFN-gamma was cell-type restricted, being seen only in macrophages and endothelial cells. In addition, phorbol ester-induced differentiation of promyelocytic HL-60 cells and promonocytic THP-1 cells rendered the gamma.1 gene inducible by IFN-gamma. The 1.0-kilobase gamma.1 cDNA sequence encoded a small predicted polypeptide of 38 amino acids and had a conserved sequence associated with rapidly turning over mRNAs. In vitro translation of the gamma.1 transcript yielded a 4,000-dalton polypeptide.

Carotid artery resection and reconstruction: clinical experience of 28 consecutive cases.

The aim of this study was to analyse the experience at a single institution in carotid artery resection with or without reconstruction performed as part of an oncological procedure or emergency haemostasis. A total of 28 patients were included in this retrospective study; 17 underwent ligation or resection of the carotid artery, and 11 underwent reconstruction of the carotid artery. The perioperative complications and surgical outcomes were recorded and analysed. Of the 17 patients with ligation or resection of the carotid artery, 4 developed neurologic deficit within 2 weeks postoperatively. Three patients with malignant tumours died 1 month (1) and 4 months (2) postoperatively. Of the 11 patients undergoing carotid reconstruction, no major cerebral complications were noted after operation. Colour Doppler showed patent vascular graft 1 year postoperatively in nine patients. Due to the higher complication rates both in short and long term with ligation or resection of the carotid artery, resection and revascularization of the carotid artery is advocated for patients with carotid artery involvement when possible.

Magnetic resonance arthrography applied to the diagnosis of intraarticular adhesions of the temporomandibular joint.

The purpose of this study is to investigate the diagnostic accuracy of magnetic resonance arthrography (MRAr) on the diagnosis of intraarticular adhesions of the temporomandibular joint (TMJ). Twenty-four patients (31 joints) diagnosed as internal disorders of TMJ were examined to establish the presence, suspicion or absence of intraarticular adhesions through MRAr according to the MRAr radiographic criteria in a blind fashion, subsequent arthroscopy were performed on all 31 joints. The findings were compared to MRAr with those of arthroscopy. The comparison data were assessed by generating receiver operating characteristic curve (ROC curve) and analyzing the area (Az) under the curve with SPSS11.0 software. The area under the ROC curve was 0.91 (0.86, 0.95), P<0.05. MRAr proves to be an excellent modality to diagnose intraarticular adhesions of TMJ.

Substantia nigra pars reticulata lesion induces preconvulsive behavior and changes in glutamate receptor gene expression in the rat brain.

The substantia nigra pars reticulata (SNpr) has been proposed to play an important role in the control of the propagation and/or the generation of epileptic seizures. Earlier studies have shown differential effects of the lesion of the SNpr on seizure genesis that demonstrated a regional difference in the anterior and posterior parts of the SNpr in preconvulsive behavior induced by unilateral reticulata injection of dopamine (DA). This study was aimed to investigate some of the underlying mechanisms of the preconvulsive behavior elicited by unilateral SNpr DA injection by the study of changes in the gene expression of glutamate receptor subunits (GluR1, GluR2 and NMDAR1) and of changes in animal behavior following coinfusion of DA and a DA D1 antagonist SCH 23390 into the SNpr. Unilateral injection of exogenous DA into the anterior region of the SNpr induced rapid and short lasting preconvulsive behavior up to wet dog shakes stage and a significant reduction of gene expression for GluR1, GluR2 and NMDAR1 subunits in rat hippocampal subfields including CA1 through CA4 and dentate gyrus (DG) at 1 day after nigral DA injection. The effect was long lasting and persisted for at least 3 weeks. Both preconvulsive behavior and downregulation of glutamate receptor subunit genes were completely blocked by simultaneous coinfusion of DA and SCH 23390. The results suggest, for the first time, that DA D1 receptor in the SNpr may mediate the nigral-involved seizure development. Glutamate desensitization, and/or selective early neuronal damage might be responsible for the downregulation of glutamate receptor subunits by transient preconvulsive activity.

Substantia nigra pars reticulata lesion facilitates kainic acid-induced seizures.

The administration of subconvulsive doses of kainic acid (5 mg/kg, intraperitoneally) to rats, with lesion of the substantia nigra pars reticulata (1 week), produced high frequency wet dog shakes and severe convulsive behavior (observed in 60% of the rats). The behavior was not observed in rats treated with kainic acid but without reticulata lesion. The results show that rats with unilateral lesion of the substantia nigra pars reticulata are more vulnerable to seizure stimuli.

Induction of preconvulsive behavior and Fos expression by dopamine-induced nigral lesion in the rat.

The substantia nigra pars reticulata (SNpr) has been proposed to play an important role in controlling the propagation and/or the generation of limbic seizures. Earlier work has shown that SN lesions have differential effects on seizure activity, suggesting that at least two discrete topographical regions mediate anticonvulsant or proconvulsant effects. The present investigation showed that exogenous dopamine (DA; 1.5-2.0 mumol) unilaterally injected into the anterior SNpr induced preconvulsive behavior (starting, immobilization, facial and mouth movements and wet-dog shakes). In addition, these rats showed Fos oncoprotein expression in the limbic system. These effects were observed in 90% of the rats with anterior SNpr DA injection. Rats with posterior SNpr injection did not show preconvulsive behavior nor Fos expression. These results show for the first time that unilateral DA lesion of the anterior portion of SNpr elicits Fos expression and preconvulsive behavior. In addition, the results suggest that lesion of the anterior and posterior regions of SNpr appear to exert different influences in the generation of preconvulsive behavior. The time course of behavior changes and Fos expression was also studied.

Lesion of the substantia nigra pars compacta downregulates striatal glutamate receptor subunit mRNA expression.

This is a study of the effect of the unilateral administration of dopamine (DA) in the pars compacta of the substantia nigra (SN) of the rat on striatal glutamate receptor subunit (GluR1, GluR2 and NMDAR1) gene expression determined by in situ hybridization. The location of the nigral lesion was determined by tyrosine hydroxylase (TH) immunohistochemistry and its extent by the striatal DA and 3,4-dihydroxyphenylacetic acid (DOPAC) concentrations. The DA-induced lesions produce significant bilateral reductions in the expression of GluR1 and NMDAR1 subunit mRNA in the medio-lateral striatum, whereas the expression of striatal GluR2 receptors was not changed. The reduction in GluR1 and NMDAR1 subunit mRNA may be the consequence of glutamatergic hyperactivity developed in the presence of a damaged nigro-striatal system and these may be associated with the genesis of some neurodegenerative diseases.

Radical resection of a Shamblin type III carotid body tumour without cerebro-neurological deficit: Improved technique with preoperative embolization and carotid stenting.

The surgical resection of a large unfavourable Shamblin type III carotid body tumour (CBT) can be very challenging technically, with many potential significant complications. Preoperative embolization aids in shrinking the lesion, reducing intraoperative blood loss, and improving visualization of the surgical field. Preoperative internal carotid artery (ICA) stenting aids in reinforcing the arterial wall, thereby providing a better dissection plane. A woman presented to our institution with a large right-sided CBT. Failure of the preoperative temporary balloon occlusion (TBO) test emphasized the importance of intraoperative preservation of the ipsilateral ICA. A combination of both preoperative embolization and carotid stenting allowed a less hazardous radical resection of the CBT. An almost bloodless surgical field permitted meticulous dissection, hence reducing the risk of intraoperative vascular and nerve injury. Embolization and carotid stenting prior to surgical resection should be considered in cases with bilateral CBT or a skull base orientated high CBT, and for those with intracranial extension and patients who have failed the TBO test.

Intralesional injection of pingyangmycin is a safe and effective treatment for microcystic lymphatic malformations in the tongue.

OBJECTIVE: To evaluate the efficacy and safety of intralesional injection of pingyangmycin for the treatment of microcystic lymphatic malformations (LMs) in the tongue. METHODS: Eighteen patients with tongue microcystic LMs were treated with intralesional injection of pingyangmycin. The concentration of the drug was 1 mg/mL with an addition of dexamethasone. Repeated injections were performed at an interval of 3­4 weeks. The results were evaluated by clinical examinations and Doppler ultrasonography scan. The follow-up period was 12 months to eight years after the last treatment and the mean follow-up time was three years. All patients received 1­8 injections (mean, 3.0 injections) for the whole course of treatment. The total dose of pingyangmycin administered was 8­64 mg (mean, 24 mg). RESULTS: Fifteen patients had complete response with no cosmetic or functional problems.Three patients with macroglossia had a reduction of 50­90% in the lesion size and needed secondary surgery. No serious complications were encountered. CONCLUSION: The results suggested that intralesional injection of pingyangmycin is an effective and safe treatment for microcystic LMs in the tongue, and can be used as the first-line treatment protocol.

Ethanol embolization of auricular arteriovenous malformations: preliminary results of 17 cases.

BACKGROUND AND PURPOSE: Because of the relatively rare and extremely varied clinical presentations, arteriovenous malformations (AVMs) involving the auriculae are technically challenging clinical entities to diagnose and, ultimately, manage. The purpose of our study was to present our initial experience of ethanol embolization in a series of 17 patients with auricular AVMs and assess the interim therapeutic outcomes of this method. MATERIALS AND METHODS: Our study group consisted of 17 patients. Transcatheter arterial embolization and/or direct percutaneous puncture embolization were performed. Pure or diluted ethanol was manually injected. Follow-up evaluation was obtained on the basis of physical examination and angiography at 3- to 4-month intervals and telephone questionnaire at 1-month intervals in all patients. RESULTS: During the 29 ethanol embolization procedures, the amount of ethanol used ranged from 4 to 65 mL. The obliteration of ulceration, hemorrhage, pain, infection, pulsation, and bruit in most of the patients was obtained. The reduction of redness, swelling, and warmth was achieved in all of the patients, and 15 of the patients achieved downstaging of the Schobinger status. According to the angiographic findings, AVMs were devascularized 100% in 3 patients, 76% to 99% in 5 patients, 50% to 75% in 6 patients, and less than 50% in 3 patients. The most common complications were reversible necrosis and blister. CONCLUSIONS: Ethanol embolization has proved efficacious and safe in the treatment of auricular AVMs and has the potential to be accepted as the primary mode of therapy in the management of these lesions.

Ethanol embolization of arteriovenous malformations of the mandible.

BACKGROUND AND PURPOSE: Absolute ethanol was reported as an effective embolization of arteriovenous malformations (AVMs), but its use to treat AVMs in the mandible is not yet well established. Here, we present our clinical experience on treatment of mandibular AVMs with absolute ethanol. MATERIALS AND METHODS: Eight consecutive patients with symptomatic AVMs of the mandible between August 2007 and September 2008 were enrolled in this study group. Among them, 6 patients underwent direct puncture embolization with absolute ethanol combined with coils, 1 patient underwent direct puncture embolization with absolute ethanol only, and the last patient had transarterial embolization with absolute ethanol alone. The use of coils decreased the flow and volume of the nidus, and then absolute ethanol embolization was directed against and obliterated the nidus completely. The procedure was performed with the patients under general anesthesia with nasal intubation, and the vital signs of the patients were constantly monitored during the injection of absolute ethanol. The total amount of absolute ethanol used per session was less than 1 mL/kg of body weight. RESULTS: A total of 11 ethanol embolizations were performed on 8 patients, including 3 sessions with transarterial microcatheterization and 8 with direct puncture embolization. A venogram and control arteriogram performed immediately after the procedure were both obtained, which documented a significant thrombosis of the lesion in all patients. Follow-up examinations revealed that oral bleeding was controlled, the expansion of the external jugular vein in 5 cases was obliterated, and satisfactory shrinkage of the facial swelling was achieved. Follow-up angiography (mean, 4.2 months) was available in 4 patients, and there was no angiographic recurrence of the lesions. There were 3 cases with minor complications. CONCLUSIONS: On the basis of our experience, treatment of AVMs in the mandible with absolute ethanol is a feasible, safe, and highly effective method.

[Function of lymphocyte beta-adrenergic receptor in depression].

Lymphocyte beta-adrenergic receptor function of 20 depressed patients and 18 healthy volunteers were measured using radioligand binding technique. It was found that the affinity and sensitivity of this receptor were significantly higher in endogenous depressed patients than those in normal controls. After the electro-acupuncture-treatment (EAT), lymphocyte beta-receptor function decreased in patients who responded well to EAT, but still not reached normal level.