[The expression and correlation analysis of TOX and inhibitory receptors on peripheral blood CD8+T cells in patients with aplastic anemia].

Objective: To investigate the expression levels of thymocyte selection related high mobility group proteins (TOX) and different inhibitory receptors in peripheral blood CD8+T cells of patients with aplastic anemia (AA), and to conduct correlation analysis. Methods: From September 2019 to November 2020, 27 AA patients in the Department of Hematology, General Hospital of Tianjin Medical University were retrospectively selected, including 21 males and 6 females, with a median age [M (Q1, Q3)] of 48 (30, 72) years. Thirty-three healthy controls, included 17 males and 16 females, with a median age of 46 (27, 69) years. The expression levels of TOX, programmed cell death receptor-1 (PD-1), T-cell immunoglobulin and mucin domain 3 (TIM-3), cytotoxic T-lymphocyte associated antigen-4 (CTLA-4), T-cell immune receptor with immunoglobulin (Ig) and immunoreceptor tyrosine-based inhibitory motif (ITIM) domain (TIGIT), perforin and granzyme B in peripheral blood CD8+T cells from AA patients and healthy controls were detected by flow cytometry. The correlation between TOX expression levels and different inhibitory receptors was analyzed using Pearson correlation analysis. Results: The expression levels of TOX, PD-1, TIM-3, CTLA-4, TIGIT, perforin, and granzyme B in peripheral blood CD8+T cells of AA patients were 47.33%(41.47%, 56.61%), (30.61±12.37)%, (39.94±10.84)%, (6.21±3.40)%, (51.45±20.21)%, (71.32±22.46)%, and (52.39±23.99)%, respectively, which were higher than those of healthy controls 27.32%(21.64%, 46.96%), (21.29±10.01)%, (21.11±3.00)%, (1.31±0.34)% (30.80±13.40)%, (46.72±22.53)%, (21.75±16.43)% (all P<0.05). The expression level of TOX in CD8+T cells was positively correlated with the expression levels of PD-1, TIM-3, CTLA-4, TIGIT, perforin, and granzyme B (r=0.49, 0.65, 0.70, 0.54, 0.58, 0.48, all P<0.05). Conclusion: The expression levels of TOX and different inhibitory receptors on peripheral blood CD8+T cells in AA patients are higher than those in the healthy control group, and the expression levels of TOX and different inhibitory receptors are positively correlated.

[Relationship of operation manner and postoperative recurrence of hepatocellular carcinoma].

Hepatocellular carcinoma (HCC) owns the high morbidity and mortality rates. Surgical resection is still the major pathway for the longer survival of HCC patients. Postoperative recurrence and metastasis have become the key impairment of prognosis of HCC patients. The relationship between tumor recurrence and surgical manner underwent by HCC patients is complicated and multiple factors are included. When the liver tumor was pressured during operation, tumor cells could be squeezed into blood flow via the broken vessels, which resulted in tumor metastasis. Besides, ischemia-reperfusion injury induced by Pringle maneuver during the liver blood blockade resulted in the immune destruction of liver and induced tumor recurrence.The destruction of physical barriers consisted of interstitial cells and normal liver cells was also a key factor for tumor recurrence. This paper summarizes the possible relationship between postoperative recurrence and surgical manner in HCC patients to provide the preventive suggestions for the postoperative recurrence of HCC patients.

[Angiopoietin-2 regulates vessels encapsulated by tumor clusters positive hepatocellular carcinoma nest-type metastasis via integrin α5ß1].

Objective: To investigate the molecular mechanism of nest metastasis in blood vessels encapsulated by tumor clusters (VETC) positive hepatocellular carcinoma (HCC). Methods: A total of 72 paraffin embedded HCC tissue samples were collected. Immunohistochemistry staining with CD34 (vascular endothelial cell marker protein) was used to observe the morphological manifestations of VETC cancer nests in primary tumors, bile duct cancerous thrombi and portal vein cancerous thrombi, and to study the characteristics of hematogenous metastasis of VETC cancer nests. Bioinformatics was used to predict the key proteins closely related to VETC cancer nest formation. Immunohistochemistry was used to detect the expression of angiogenin-2 (Ang-2), integrin α5, Integrin ß1, and cyclooxygenase-2 (COX-2) proteins in HCC. Transwell cell migration assay was used to detect the effect of Ang-2/integrin α5ß1 protein on the migration ability of endothelial cells and HCC cells. Western blotting was used to detect the effect of Ang-2/integrin α5ß1 protein on the activity of focal adhesion kinase (FAK) protein. Results: Of the collected HCC specimens, 27 cases (27/72) were VETC (+), including 3 cases with biliary duct cancerous thrombus, 5 cases with portal vein cancerous thrombus, and 3 cases with both biliary duct cancerous thrombus and portal vein cancerous thrombus. VETC (+) HCC could metastasize to portal vein, bile duct, and liver in the form of cancer nest, and the nests retain their intact structure. Ang-2, integrin α5 and integrin ß1 were overexpressed in tumor cells and endothelial cells of VETC (+) HCC nests, while COX-2 was only overexpressed in tumor cells of VETC (+) HCC nest. Ang-2 could promote the migration of HCC cell [(121±12) vs (186±11), P<0.01] and endothelial cells [(81±7) vs (163±14), P<0.01]. Integrin α5ß1 activation antagonist ATN-161 could significantly block the ability of Ang-2 to promote the migration of HCC cells [(185±10) vs (135±9), P<0.05] and endothelial cells [(156±14) vs (103±6), P<0.05]. ATN-161 could significantly block the phosphorylation of FAK in HCC and endothelial cells induced by Ang-2. Conclusions: VETC (+) HCC could metastasize as a whole in a nested form, and possesses a specific regulatory protein. Ang-2/α5ß1/FAK might be potential protein targets in the treatment of VETC (+) HCC nest-type metastasis.

[Study on the prognostic value of preoperative anti hepatitis B virus therapy in patients with hepatocellular carcinoma complicated with microvascular tumor thrombus and establishment of survival prediction model].

Objective: To explored the effect of preoperative antiviral therapy on the prognosis of microvascular tumor thrombi patients, and to established a prognostic prediction model for these patients after radical resection of liver cancer. Methods: The clinicopathological and survival data of hepatocellular carcinoma patients with microvascular tumor thrombus who underwent radical resection in the Third Affiliated Hospital of Sun Yat-sen University from January 1, 2013 to December 31, 2015 were retrospectively collected. Kaplan-Meier method was used to calculate the survival curve, and log-rank test was used to compare the prognosis of patients with and without antiviral treatment before operation. Univariate and multivariate Cox proportional hazard regression model was used to screen predictive factors. R software was used to make predictive nomogram, and discrimination and calibration degree were used to evaluate the prediction model. Results: Among all 153 patients, 22 were female and 131 were male, aged (51.3±11.7) years. The preoperative antiviral therapy significantly improved overall survival and recurrence-free survival (χ2=41.423, 54.389; both P<0.001). According to the results of multivariate and regression analysis, preoperative antiviral therapy (HR=0.301,95%CI:0.171-0.532,P<0.001), alpha fetoprotein (HR=1.226,95%CI:1.157-1.776,P=0.032) and tumor size (HR=1.008,95%CI:1.001-1.016,P=0.02) were important prognostic factors for overall survival. The area under curve value of 3-year survival prediction model was 0.749(95%CI: 0.712-0.782), and that of 5-year survival prediction model was 0.755(95%CI: 0.724-0.793), with good calibration. Conclusions: Preoperative anti hepatitis B virus(HBV) therapy can significantly improve the prognosis of patients with hepatocellular carcinoma complicated with microvascular tumor thrombus, we develope the prediction models of 3-year and 5-year survival rate that can improve the reference for clinical work and benefit patients.

Using real-time sound touch elastography to monitor changes in transplant kidney elasticity.

AIM: To use sound touch elastography (STE) to assess the changes of renal cortex among different complications following renal transplantation. MATERIALS AND METHODS: A total of 31 patients with renal dysfunction after renal transplantation underwent an ultrasound-guided biopsy for pathological examination with conventional and STE ultrasound. The maximum elastic modulus (Emax) was determined, and the biopsy specimen was evaluated for evidence of significant differences among four different complications: drug-induced renal damage, acute rejection, chronic allograft nephropathy (CAN), and BK virus (BKV) nephropathy. Receiver operator characteristics were used to compare the diagnostic efficacy of STE ultrasound according to the pathological results. RESULTS: The quantitative index Emax of the STE technique was statistically significant among the four different complications (p<0.05). The distribution of the magnitude of Emax in the renal cortex was BKV nephropathy > CAN > acute rejection > drug induced renal damage. The renal cortex Emax was statistically different for the severity of renal fibrosis and tubular atrophy (p<0.05). CONCLUSION: Each of the four different complications of transplantation influenced the Emax of the renal cortex differently. Emax can be used to assess the severity of renal fibrosis and tubular atrophy.

[Research progress in the Hepatobiliary Surgery operation of hepatic hilar plate system].

An estimate of about 50% of new liver cancer cases worldwide occur in China every year.Surgical resection is still the major treatment choice for longer survival of patients with hepatocellular carcinoma. Blocking hepatic blood flow and reducing intraoperative bleeding ensure the success of the operation. Anatomic separation of hepatic hilar region is the precondition of hepatic inflow occlusion. The hepatic hilar plate system involves a thick layer of connective tissue covering the hepatic inflow ducts of hepatic hilar region. The descending part of hilar plate assists in reducing the anatomical difficulty of the hepatic hilar region. The "forth porta hepatis" that is hidden in the hepatic hilar plate system involves the accumulation area of "short hepatic portal veins" .The communicating branch vessels between the hepatic inflow vessels form the anatomical basis in reducing the indocyanine green fluorescence stain effect.The relatively fixed position of the hepatic portal plate is considered as a positioning marker for accurate liver resection. The intrahepatic Glisson sheath is connected with thick connective tissue of the hepatic portal panel system, and is regarded as the physical barrier in limiting the proliferation and hypertrophy of hepatocytes and continuation of hepatic portal panel system in the liver.This paper summarizes the anatomy and application of hepatic hilar plate system during hepatobiliary surgery.

[Proton nuclear magnetic resonance spectroscopy recognition of metabolic patterns in fecal extracts for early diagnosis of colorectal cancer].

Objective: To characterize the metabolic " fingerprint" of fecal extracts for diagnosis of early-stage colorectal cancer(CRC)using proton nuclear magnetic resonance spectroscopy(1H-NMR)-based metabolomics coupled with pattern recognition. Methods: From January 2014 to December 2014, we collected fecal samples at the Second Affiliated Hospital of Shantou University Medical College, from 25 patients with colorectal adenomas(CR-Ad), 20 with stage Ⅰ/Ⅱ CRC, and 32 healthy controls(HCs). The patients were diagnosed by histopathology. No subjects had any complicating diseases. HCs showed no abnormalities from blood tests, endoscopic examination, diagnostic imaging, and/or medical interviews. We excluded participants who used antibiotics, NSAIDS, statins, or probiotics within two months of study participation, and any patients who underwent chemotherapy or radiation treatments prior to surgery. We used orthogonal partial least-squares-discriminant analysis(OPLS-DA)for pattern recognition(dimension reduction)on 1H-NMR processed data(1H frequency of 400.13 MHz), to find metabolic differences among CR-Ad, carcinoma and HC fecal samples; and receiver operating characteristic(ROC)analysis to determine the diagnostic value of the fecal metabolic biomarkers. Results: Fecal samples were collected from 20 patients with Stage Ⅰ/Ⅱ CRC(11 M, 9 F, median age(52±13)years), 25 with CR-Ad(14 M, 11 F, median age(53 ± 11)years)and 32 HCs(15 M, 17 F, median age(53 ± 14)years). OPLS-DA clearly distinguished CR-Ad and stage Ⅰ/Ⅱ CRC from HC samples, based on their metabolomic profiles. Relative signal intensities in HCs were significantly lower than in the cancer patients for butyrate(HC: 23.0±6.0; CR-Ad: 18.0±5.0; CRC: 14.0±6.0; Z=-2.07, P=0.008), acetate(HC: 45.0±11.0; CR-Ad: 31.0±11.0; CRC: 24.0±8.0; Z=- 2.32, P=0.011), propionate(HC: 26.0 ± 7.0; CR-Ad: 22.0 ± 6.0; CRC: 19.0 ± 5.0; Z=- 2.43, P=0.032), glucose(HC: 37.0±7.0; CR-Ad: 31.0±7.0; CRC: 26.0±8.0; Z=-2.07, P=0.044)and glutamine(HC: 4.5±2.0; CR-Ad: 4.9 ± 1.0; CRC: 5.4 ± 1.0; Z=2.21, P=0.044). However, relative signal intensities in HCs were significantly higher than in patients for lactate(HC: 4.8±1.0; CR-Ad: 6.9±2.0; CRC: 4.8± 1.0; Z=2.02, P= 0.038), glutamate(HC: 3.2 ± 2.0; CR-Ad: 4.9 ± 1.0; CRC: 3.2 ± 2.0; Z=2.21, P=0.044)and succinate(HC: 12.0±2.0; CR-Ad: 15.0±3.0; CRC: 12.0± 2.0; Z=2.25, P=0.011). Among the potential biomarkers, acetate at 1.92 ppm, and succinate at 2.41 ppm displayed relatively high area under ROC, with sensitivity and specificity both >90%, to distinguish early-stage CRC patients from HCs. Conclusion: Fecal metabolic profiles distinguish of HCs from patients with CRC patients, even in the early stages(stage Ⅰ/Ⅱ), highlighting the potential of NMR-based fecal metabolomic fingerprinting as tools for early CRC diagnosis.

Serological comparison of antibodies to avian influenza viruses, subtypes H5N2, H6N1, H7N3 and H7N9 between poultry workers and non-poultry workers in Taiwan in 2012.

In Taiwan, avian influenza virus (AIV) subtypes H5N2, H6N1 and H7N3 have been identified in domestic poultry, and several strains of these subtypes have become endemic in poultry. To evaluate the potential of avian-to-human transmission due to occupational exposure, an exploratory analysis of AIV antibody status in poultry workers was conducted. We enrolled 670 poultry workers, including 335 live poultry vendors (LPVs), 335 poultry farmers (PFs), and 577 non-poultry workers (NPWs). Serum antibody titres against various subtypes of viruses were analysed and compared. The overall seropositivity rates in LPVs and PFs were 2·99% (10/335) and 1·79% (6/335), respectively, against H5N2; and 0·6% (2/335) and 1·19% (4/335), respectively, for H7N3 virus. Of NPWs, 0·35% (2/577) and 0·17% (1/577) were seropositive for H5N2 and H7N3, respectively. Geographical analysis revealed that poultry workers whose workplaces were near locations where H5N2 outbreaks in poultry have been reported face greater risks of being exposed to viruses that result in elevated H5N2 antibody titres. H6N1 antibodies were detected in only one PF, and no H7N9 antibodies were found in the study subjects. Subclinical infections caused by H5N2, H6N1 and H7N3 viruses were thus identified in poultry workers in Taiwan. Occupational exposure is associated with a high risk of AIV infection, and the seroprevalence of particular avian influenza strains in humans reflects the endemic strains in poultry in this region.

Latent transforming growth factor-ß binding proteins (LTBP-1 and LTBP-2) and gingiva keratinization.

OBJECTIVE: Transforming growth factor-beta (TGF-ß) proteins are involved in epithelial keratinization. The major function of latent TGF-ß binding proteins (LTBPs) is modulating TGF-ß activity. However, whether LTBP-1 and LTBP-2 play roles in gingiva keratinization remains unclear. MATERIALS AND METHODS: Human keratinized gingiva and non-keratinized alveolar mucosa were processed for LTBP-1, LTBP-2, cytokeratin-1 (K1), cytokeratin-4 (K4), and TGF-ß immunohistochemical (IHC) staining. Porcine heterotopically transplanted connective tissues and newly grown epithelia were harvested for IHC staining. The expression levels of LTBP-1 and LTBP-2 were compared between differentiated and undifferentiated human normal oral keratinocytes (hNOK). The expression of LTBP-1 and LTBP-2 was knocked down in a cell line (OEC-M1) to evaluate the effects on the expression of K1, K4, and involucrin (INV). RESULTS: In human and porcine specimens, LTBP-2 expression patterns distinguished keratinized and non-keratinized oral epithelia. Western blotting results showed that K1, LTBP-1, and INV proteins were upregulated in differentiated hNOK. In OEC-M1 cells, LTBP-2 knockdown resulted in upregulated the expression of K1 and INV and downregulated the expression of K4. LTBP-1 knockdown resulted in opposite effects. CONCLUSION: The expression patterns of LTBP-2 differ in keratinized gingiva and non-keratinized mucosa. LTBP-1 and LTBP-2 are involved in the keratinization of oral epithelium; however, the underlying mechanism remains to be elucidated.

Ischemia-reperfusion injury in an aortic dissection patient.

Aortic dissection is a life-threatening emergency. Well-established risk factors include systemic hypertension, hereditary connective tissue diseases (Marfan syndrome and Ehlers-Danlos syndrome), coarctation of the aorta, bicuspid aortic valve, aortitis, and arch hypoplasia. Ischemia of the viscera, the kidneys, the spinal cord, or the lower extremities due to malperfusion constitutes life-threatening complications that have to be considered in the treatment strategy.We report a rare case of symptomatic ischemia of the lower extremities due to aortic dissection. This case demonstrates that the treating physician needs to be vigilant for ischemia reperfusion injuries such as osteofascial compartment syndrome and acute renal failure in aortic dissection.

Quality of life of Chinese urologists: a cross-sectional study using WHOQOL-BREF.

OBJECTIVES: In recent years, Chinese hospital settings are under violent threats. The exact status of quality of life of Chinese doctors under these disastrous situations remains obscure. The aim of this study was to assess the quality of life of Chinese urologists and analyse its potential affecting factors. DESIGN: Cross-sectional survey. SETTING: Beijing, China. PARTICIPANTS: Overall, 1000 participants from more than 30 areas of China, who participated in the 20th National Urology Conference in Beijing in 2013, were surveyed. The brief version of the World Health Organization Quality of Life (WHOQOL-BREF) Chinese version was used to assess the quality of life among these urologists. The relationship between quality of life and the affecting factors was analysed. RESULTS: Of the 1000 questionnaires, 856 were completed and returned, and 708 questionnaires were valid for analysis. Approximately 46% of the respondents came from provincial capitals, 54.2% of them felt stress from medical environment, while 76.0% felt stress from research work, and 85.3% from promotion. Cronbach's α coefficient of the instrument was 0.825, Kaiser-Meyer-Olkin measure was 0.841, and P value of Bartlett's sphericity was <0.001. The results of binary logistic regression indicated gender, work years, and medical environment as potential affecting factors of quality of life only influenced one domain. In contrast, research work and promotion influenced three domains of the WHOQOL-BREF. CONCLUSIONS: The study indicated that the WHOQOL-BREF may be a reliable and valid tool to assess quality of life of Chinese urologists. In China it is true that the deteriorative medical environment negatively affects medical practice according to previous studies, and policies are recommended to improve the situation. Nevertheless, we should not be too pessimistic about it, as in today's context research work and promotion may be the most extensive and significant affecting factors on doctors' quality of life.

Triptolide inhibits the multidrug resistance in prostate cancer cells via the downregulation of MDR1 expression.

Triptolide (TPL) is a diterpenoid triepoxide derived from the Chinese herb Tripterygium wilfordii and possesses anti-tumor activity against a range of cancer cells. However, the effect of TPL on prostate cancer cells and its potential to overcome multidrug resistance (MDR) have not been explored. Therefore, in this study we used prostate cancer cell line DU145 as the experimental model and established DU145/ADM cell line resistant to adriamycin (ADM). Our results showed that TPL inhibited the proliferation and induced the cell cycle arrest and apoptosis of DU145 cells in a dose and time dependent manner. TPL decreased the levels of Cyclin D1 and anti-apoptotic protein Bcl-2, and increased the levels of pro-apoptotic proteins Fas and Bax. Furthermore, we found that TPL restored the sensitivity DU145/ADM cells to ADM in a dose dependent manner, and this was accompanied by the inhibition of MDR1 expression at both mRNA and protein levels. Taken together, these results provide strong evidence that TPL overcomes MDR in prostate cancer cells by downregulating MDR1 expression, and suggest that TPL is a promising agent for prostate cancer therapy, especially for chemoresistant prostate cancer.

Surveillance of avian influenza A(H7N9) virus infection in humans and detection of the first imported human case in Taiwan, 3 April to 10 May 2013.

On 3 April 2013, suspected and confirmed cases of influenza A(H7N9) virus infection became notifiable in the primary care sector in Taiwan, and detection of the virus became part of the surveillance of severe community-acquired pneumonia. On 24 April, the first imported case, reported through both surveillance systems, was confirmed in a man returning from China by sequencing from endotracheal aspirates after two negative throat swabs. Three of 139 contacts were ill and tested influenza A(H7N9)-negative.

Verticillium Wilt of Redbud in China Caused by Verticillium dahliae.

Chinese redbud (Cercis chinensis Bunge), a member of the Fabaceae, is an important ornamental plant native to China with reported desirable medicinal effects, including stimulating blood circulation, detumescence, and detoxification (1). In October 2011, wilt symptoms of gradual leaf yellowing, wilting, scorching (marginal browning), and twig dieback were observed on plants in Yangling, Shaanxi, China. The incidence of diseased plants was about 20% in two main areas (about 20 ha in Zijingshan Park in Zhengzhou, Henan Province; and about 3,000 ha in Taiping National Forest Park in Xi'an, Shaanxi Province). Wilted leaves of diseased plant senesced and died, but defoliation was not observed. Brown discoloration was observed in vascular tissues of petioles, twigs, and stems of diseased plants, sometimes in a ring pattern. The symptoms were often restricted to the lower part of the tree or a few branches. To identify the causal agent, six twigs (each approximately 50 mm in diameter and 10 cm long) sampled from an infected tree in Yangling were rinsed in running water, surface-sterilized with 75% ethanol for 2 min, rinsed in sterilized water three times, dried, cut into 1 cm long segments, and the segments put onto potato dextrose agar (PDA) medium. A fungal isolate was recovered from diseased vascular tissues of each sample when cultured on PDA in the dark at 25°C. After 5 days, colonies changed from white to black as a result of production of microsclerotia. Microscopic observation revealed that conidiophores were hyaline and verticillate, with three to four phialides at each node. Conidia were ellipsoidal, hyaline, single-celled, and 2.5 to 7.5 × 1.25 to 4.5 µm. On the basis of these morphological characteristics, the fungus was identified as Verticillium dahliae (3). To prove Koch's postulates, the roots of 10 healthy, 1-year-old C. chinensis plants were each irrigated in a greenhouse with 50 ml of a conidial suspension (1.0 × 107 spores/ml) of an isolate recovered from an infected plant (2); five control plants were inoculated similarly with sterilized water. Fifteen days after inoculation, the same wilt symptoms observed on the original plants had developed on 9 of the 10 inoculated plants, whereas the control plants remained healthy. The pathogen was recovered 15 days after inoculation by isolating from petiole and stem tissues of symptomatic plants, but was not isolated from the control plants. The internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2) of the nuclear ribosomal DNA was PCR-amplified with primers ITS1 and ITS4 (4), and sequenced. BLAST analysis of the ITS sequence (GenBank Accession No. AB735536) showed 100% homology with that of an isolate of V. dahliae (FJ572050). To our knowledge, this is the first report of Verticillium wilt on C. chinensis in China. References: (1) Y. Li et al. J. Integr. Plant Biol. 47:1021, 2005. (2) H. A. Melouk and C. E. Horner. Phytopathology 65:767, 1975. (3) G. F. Pegg and B. L. Brady. Verticillium Wilts, CABI Publishing, Oxford, UK, 2002. (4) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, CA, 1990.

Genitourinary toxicity of ketamine.

Ketamine is a relatively new recreational drug used by youngsters in recent decades. Its toxic effects on the genitourinary system were first reported in 2007, and now attract extensive attention from urologists, pharmacologists, and toxicologists all over the world. As many front-line health professionals and medical social workers are still unaware of this new clinical entity and an increasing number of the drug users seek help for urological symptoms, this mini-review aimed to summarise the clinical features and possible mechanisms of ketamine-induced genitourinary toxicity. By raising public awareness of these toxic effects, the authors hope that the contents of this review will be widely disseminated not only to medical professionals, but also to relevant government departments and the general public.

First Report of Wilt on Alfalfa in China Caused by Verticillium nigrescens.

Alfalfa (Medicago sativa Linn.), widely grown throughout the world, is an important perennial forage crop. It is high in protein and digestible fiber and is an excellent source of several vitamins (A, D, E, and K) and minerals for beef cattle, horses, sheep, goats, and even humans (2). Wilt symptoms on alfalfa were observed during a disease survey in Yangling, Shaanxi, China in 2009. Symptoms included discoloration, shortened internode, and plant death. However, the vascular tissue of diseased alfalfa plants did not exhibit discoloration and typical "V" symptoms of Verticillium albo-atrum infection. Eleven fungal isolates were obtained from diseased alfalfa plants in Yangling by a tissue isolation method (1). Isolates were cultured on Czapek Dox Agar (CDA; pH 7.2) slants at 22 ± 1°C in darkness. Colonies on CDA plates were whitish and cream-white when viewed from the underside, later becoming dark gray due to the formation of gray or dark brown chlamydospores in single or in short chains. DNA was extracted from each isolate and the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA (rDNA) was amplified and sequenced using primers ITS-1F and ITS4. The 11 isolates were divided into five groups based on their in vitro morphological characters. A single isolate from each of the five groups was chosen for ITS sequencing. All five isolates had the same ITS sequence (GenBank Accession No. AB551216). On the basis of the ITS sequence and morphology (4), these isolates were identified as V. nigrescens Pethyhr. (recently renamed as Gibellulopsis nigrescens). Five representative isolates were used to fulfill Koch's postulates. Alfalfa seeds (cv. Cossack) were surface sterilized with 75% ethanol for 5 min, allowed to dry, and planted into cow dung compost that had been autoclaved at 160°C for 2 h. Plants were cultivated under controlled greenhouse conditions at 23 to 25°C with a photoperiod of 14 h. Inoculum was prepared by comminuting 15-day-old cultures and sterile deionized water into a suspension of mycelial fragments and conidia (105 to 106 CFU/ml) in a blender. Seedlings (four-leaf stage) were inoculated by immersing roots in the inoculum suspension for 60 min (3). Each isolate was inoculated onto 30 seedlings, six in each pot; another 30 seedlings were soaked with sterile deionized water for 60 min as a control. After 20 days in the greenhouse, all inoculated plants exhibited wilt symptoms similar to the original wilt symptoms observed on diseased alfalfa plants. In contrast, none of the control plants showed wilt symptoms. The pathogen was reisolated from all diseased plants and confirmed to the original ones. To our knowledge, this is the first report of V. nigrescens infecting M. sativa in China, indicating V. nigrescens as one possible important pathogen of alfalfa. References: (1) O. D. Dhingra and J. B. Sinclair. Basic Plant Pathology Methods. CRC Press, Boca Raton, FL, 1995. (2) D. Jasjeet et al. J. Adv. Sci. Res. 2:50, 2011. (3) H. A. Melouk and C. E. Horner. Phytopathology 64:1267, 1974. (4) R. Zare et al. Nova Hedwigia 85:463, 2007.

Electron microscopy investigations of V defects in multiple InGaN/GaN quantum wells and InGaN quantum dots.

The mechanism of high emission of InGaN-based multiple quantum wells, which exhibit exceptionally high light emission efficiency despite their high defect density, is still not fully understood. Here, we deal with this problem, showing the details of structure and formation of V defects in the multiple quantum wells and reviewing interpretations proposed so far. Then, we show a structural investigation of three-dimensional high-density quantum dots, fabricated instead of quantum wells in the active layer. The shape and size of the InGaN quantum dots and the SiN(x) masks for the growth of the dots have been revealed using high-angle annular dark field scanning transmission electron microscopy, energy dispersive X-ray spectroscopy nanoanalysis and high-resolution transmission electron microscopy.

[Healthy China Strategy and schistosomiasis control].

This paper summarizes the changes in the policy associated with schistosmiasis control in the new era, analyzes the background of Health China Strategy and its association with the current schistosomiasis control program in China, describes several schistosomiasis control models and proposes some suggestions responding to the challenges in current schistosomiasis control program of China, so as to provide insights into the development of the effective control strategy for schistosomiasis.

Higher liver stiffness scores are associated with early kidney dysfunction in patients with histologically proven non-cirrhotic NAFLD.

AIM: The association between Liver fibrosis (LF), as assessed by either histology or Liver stiffness measurement (LSM), and the presence of Early kidney dysfunction (EKD) was investigated in this study, as was also the diagnostic performance of LSM for identifying the presence of EKD in patients with Non-alcoholic fatty liver disease (NAFLD). MATERIALS AND METHODS: A total of 214 adults with non-cirrhotic biopsy-proven NAFLD were recruited from two independent medical centres. Their histological stage of LF was quantified using Brunt's criteria. Vibration-controlled Transient elastography (TE), using M-probe (FibroScan®) ultrasound, was performed in 154 patients and defined as significant when LSM was≥8.0kPa. EKD was defined as the presence of microalbuminuria with an estimated glomerular filtration rate≥60mL/min/1.73 m2. Logistic regression modelling was used to estimate the likelihood of having EKD with NAFLD (LSM-EKD model). RESULTS: The prevalence of EKD was higher in patients with vs without LF on histology (22.14% vs 4.82%, respectively; P<0.001) and, similarly, EKD prevalence was higher in patients with LSM≥8.0kPa vs LSM<8.0kPa (23.81% vs 6.59%, respectively; P<0.05). The area under the ROC curve of the LSM-EKD model for identifying EKD was 0.80 (95% CI: 0.72-0.89). LF detected by either method was associated with EKD independently of established renal risk factors and potential confounders. CONCLUSION: LF was independently associated with EKD in patients with biopsy-proven NAFLD. Thus, TE-measured LSM, a widely used technique for quantifying LF, can accurately identify those patients with NAFLD who are at risk of having EKD.

Efficacy of sildenafil on erectile dysfunction of newlyweds.

To explore the efficacy of sildenafil on erectile dysfunction (ED) of newlyweds, the author studied 60 outpatients within a month of marriage, who suffered from sexual intercourse (SI) failure caused by ED and showed no improvement after receiving sex education and psychological consultation. The patients were given oral sildenafil, 100 mg for the first and second times, 50 mg for the third and fourth times, no more than once every day, with a 1- to 3-day break between every two times. Four times of sildenafil administration formed one course of treatment. Sildenafil was taken 1 h before SI and was aided with adequate sexual stimulation. The rates of successful SI due to improved erection during and after a course of sildenafil treatment were 93.3% (56/60) and 85% (51/60), both P > 0.05. In the groups with one and more than one SI failure the successful SI rates after a sildenafil treatment course were 93.1% (27/29) and 77.4% (24/31), both P > 0.05. Oral sildenafil with psychological therapy in the treatment of ED of newlyweds proves to be effective in restoring the patients' sexual function and relieving their mental pressure or stress.