[Role of alternative activation of macrophages in hookworm therapy for inflammatory diseases: a review].

As a type of highly plastic innate immune cells, macrophages may be differentiated into M1 and M2 macrophages upon different stimuli, and M2 macrophages are involved in immune regulation, tissue remodeling and regeneration, and wound healing. Previous epidemiological studies have shown a significant negative correlation between the prevalence of helminth infections and the incidence of inflammatory diseases, such as allergy and autoimmune diseases. As a common type of intestinal helminths, hookworm infection may trigger high levels of type II host immune responses, with alternative activation of macrophages, which are effective to inhibit the development and progression of inflammatory diseases. This review summarizes the advances in alternative activation of macrophages in hookworm therapy for inflammatory diseases.

Silencing of linc00337 inhibits proliferation, cell cycle progression, migration, and invasion of colorectal cancer cells through the MEK/ERK pathway.

OBJECTIVE: To explore the expression and biological functions of linc00337 in colorectal cancer (CRC), as well as its underlying mechanism. PATIENTS AND METHODS: The relative expression of linc00337 in 47 cases of CRC tissues and cells was detected via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). The si-linc00337 interference sequences were designed and transiently transfected into CRC cells. The interference efficiency was detected via qRT-PCR. Regulatory effect of linc00337 on proliferation of CRC cells was detected via colony formation assay. Cell cycle distribution and apoptosis rate after interference in linc00337 expression were determined using flow cytometry. Moreover, the effects of linc00337 knockdown on cell migration and invasion were detected using transwell assay. At last, the effect of si-linc00337 on the MEK/ERK signaling pathway was detected using Western blotting. RESULTS: The results of qRT-PCR showed that among the 47 cases of CRC tissues, the expression of linc00337 was up-regulated in 40 cases. Similarly, it was highly expressed in CRC cell lines. The results of colony formation assay manifested that cell proliferation declined after interference in linc00337 expression. The results of flow cytometry and transwell assay showed that interference in linc00337 expression arrested the cell cycle in G1/G0 phase, increased the apoptosis rate, and inhibited the invasion and migration of CRC cells. According to the results of Western blotting, expressions of molecular markers in the MEK/ERK pathway after interference in linc00337 expression were significantly changed. CONCLUSIONS: Linc00337 is up-regulated in CRC tissues and cells. Interference in linc00337 expression can inhibit cell proliferation, migration, and invasion and promote apoptosis through the MEK/ERK pathway.

MiR-652-3p promotes bladder cancer migration and invasion by targeting KCNN3.

OBJECTIVE: Increasing evidence indicated that microRNAs (miRNAs) are crucial regulators for cancer development. Bladder cancer (BCa) is a major threat to human health. The aim of this study was to analyze the roles of miR-652-3p in BCa, and to explore the associated mechanisms. MATERIALS AND METHODS: MiR-652-3p expression in BCa cell lines was explored using Real Time-quantitative Polymerase Chain Reaction (RT-qPCR) method. MiR-652-3p expression level in BCa tissues was explored at StarBase. Cell Counting Kit-8 (CCK-8) assay, wound-healing assay, and transwell invasion assay were conducted to investigate the biological roles of miR-652-3p. The underlying mechanisms of miR-652-3p in NSCLC were investigated using luciferase activity reporter assay and rescue experiments. RESULTS: We showed that miR-652-3p expression level was upregulated in both BCa tissues and cell lines. The knockdown of miR-652-3p significantly inhibited BCa cell proliferation, migration, and invasion in vitro. Moreover, we showed that potassium intermediate/small conductance calcium-activated channel, subfamily N, member 3 (KCNN3) was a functional target for miR-652-3p. Besides, the expression of KCNN3 in BCa tissues was negatively correlated with miR-652-3p. CONCLUSIONS: Collectively, these results showed that miR-652-3p could promote BCa cell proliferation, migration, and invasion via directly regulating KCNN3, which may provide a novel therapeutic target for BCa treatment.

[Brain protection strategy and effectivity in pulmonary thromboendarterectomy].

Objective: To summarize the experience and effectivity of brain protection in 25 patients who suffered from chronic thromboembolic pulmonary hypertension (CTEPH) and received pulmonary thromboendarterectomy (PTE) under deep hypothermic circulatory arrest. Methods: Retrospective analysis of 25 PTE surgeries in our center from December 2016 to August 2018. All cases were completed underdeep hypothermic circulatory arrest. Standard brain protections were strictly executed, including: balanced and controlled extracorporeal circulation cooling, cerebral oxygen saturation (rSO(2)) monitoring, strictly control of circulatory arrest time, and etc. The neurological adverse events during the perioperative period were recorded and statistically analyzed, and the intelligence level and cognitive function of the patients were evaluated by MMSE scale and MoCA scale before surgery and discharge. Results: All the 25 patients successfully completed the surgery, and 1 patient (4%) died of postoperative infection. The mean pulmonary arterial pressure decreased from (52.9±16.7) mmHg before surgery to (23.6±8.1) mmHg immediately after surgery (t=10.01, P<0.01), and(20.7±7.9) mmHg at 3 months follow-up (t=10.73, P<0.01). Pulmonary vascular resistance decreased from 975.4 (788.6-1 292.8) dyn·s·cm(-5) to 376.1 (283.6-565.5) dyn·s·cm(-5) (Z=5.34, P<0.01). Neurological complications occurred in 3 patients during the perioperative period, including 2 patients with hypoxic encephalopathy, and 1 patient with cerebral hemorrhage. All 3 patients fully recovered before discharge. Univariate analysis showed that the duration of rSO(2)<40% and the maximum decrease rate of rSO(2) from baseline were significantly correlated with postoperative neurological damage. Multivariate analysis showed only time of rSO(2)<40% was significantly correlated with postoperative neurological damage. There was no significant difference in MMSE and MoCA score before and after surgery (P>0.05). Conclusions: Adequate brain protection measures are essential to reduce the neurological complications of PTE surgery. Real-time intraoperative monitoring of rSO(2) and strict control of circulatory arrest time can further reduce the occurrence of neurological damage.

[Analysis of risk factors for cerebrovascular complications after carotid endarterectomy].

Objective: To evaluate the related risk factors of cerebrovascular complications after carotid endarterectomy (CEA) and to improve the efficacy of CEA in the treatment of ischemic stroke. Methods: The clinical data of 295 patients with atherosclerotic carotid artery stenosis who underwent CEA in the Department of Vascular Surgery, Beijing Anzhen Hospital, Capital Medical University from January 2013 to March 2017 were retrospectively analyzed. Results: As the results of the single-factor analysis of logistics, severe lower limb artery stenosis (RR=5.667, P=0.017), systolic blood pressure before the carotid artery clamping (RR=6.659, P=0.010), diastolic blood pressure before the carotid artery clamping (RR=3.981, P=0.046), stump pressure (RR=5.359, P=0.021), diastolic blood pressure after surgery (RR=9.550, P=0.002), diastolic blood pressure of the first day after surgery (RR=7.932, P=0.005) were influencing factors of postoperative cerebrovascular complications after CEA. The results of multi-factor analysis of logistic regression indicated that diastolic blood pressure before the carotid artery clamping (RR=0.953, P=0.024) and stump pressure to basic systolic blood pressure index (SSI)>0.25 (RR=0.086, P=0.049) were independent risk factors for postoperative cerebrovascular complications after CEA. Conclusion: Systolic blood pressure before carotid artery clamping and SSI>0.25 are independent risk factors for postoperative cerebrovascular complications after CEA. Close follow-up and drug treatment for patients after CEA might be beneficial to reduce postoperative carotid artery restenosis.

[Effects of Blood-brain Barrier and Simulated metabolic system on Apoptosis of SH-SY5Y Induced by Acrylamide in Vitro].

Objective: To evaluate the effect of acrylamide on the apoptosis of nerve cells by integral cell modelling in vitro which simulates the barrier effect and metabolic micro-environment. Methods: A non-contact and co-cultured in vitro blood brain barrier (BBB) model was established by using human umbilical vein endothelial cells (HUVEC) and rat glioma cells (C6) . The trans-endotheilal electrical resistance (TEER) and horseradish peroxidase (HRP) tracer effects were measured to verify the tight connectivity and permeability of the established BBB model. An integrate discrete multiple organ cell co-culture (idMOC) model was established by inoculating the human renal cortical proximal tubular epithelial cells (HK-2) , human normal hepatocytes (L-02) and human neuroblastoma cells (SH-SY5Y) into the self-made multi-organ plate for co-culturing. Then the model was verified by observing the growth curve of various tissue cells under co-culturing or culturing individually. SH-SY5Y cells were exposed to different concentrations of acrylamide directly and indirectly (through BBB model and idMOC model) . The changes of cell apoptosis rate were analyzed by flow cytometry to explore the impact of model on Acrylamide (ACR) injury of typical neurotoxic agents. Results: HUVEC cells can form a wide range of close-connected complex and then inhibit the external electric field under the cross-endothelial movement, and the mean was lower than that of endothelial cell culture group at 4, 5 and 6 days (P<0.05) ; After 20 min, the penetration rate of HRP in the co-culture group was less than that in the individual culture group, and the difference was statistically significant (P<0.05) , indicating that the barrier function of the co-culture group was higher than that of the individual culture group. All cells can exchange substances through the exchange hole of the culture plate, the cells grow well and there was no obvious death. The growth curve in individual culture group and co-culture group were basically the same, the difference was not statistically significant (P>0.05) . Under the condition of different concentrations of ACR (140, 270 g/ml) , compared with the direct exposure group, the apoptosis rate of the BBB model and the idMOC model were significantly decreased, and the difference was statistically significant (P<0.05) . Conclusion: Based on HUVEC cells and C6 cells co-culture system, a blood-brain barrier model in vitro was established and based on co-culture of HK-2, L-02 and SH-SY5Y, the idMOC model was established. The toxicity and toxic action characteristics of ACR on SH-SY5Y cells were evaluated by validation tests.

[Distribution and drug resistance of pathogens at hematology department of Jiangsu Province from 2014 to 2015: results from a multicenter, retrospective study].

Objective: To describe the distribution and drug resistance of pathogens at hematology department of Jiangsu Province from 2014 to 2015 to provide reference for empirical anti-infection treatment. Methods: Pathogens were from hematology department of 26 tertiary hospitals in Jiangsu Province from 2014 to 2015. Antimicrobial susceptibility testing was carried out according to a unified protocol using Kirby-Bauer method or agar dilution method. Collection of drug susceptibility results and corresponding patient data were analyzed. Results: The separated pathogens amounted to 4 306. Gram-negative bacteria accounted for 64.26%, while the proportions of gram-positive bacteria and funguses were 26.99% and 8.75% respectively. Common gram-negative bacteria were Escherichia coli (20.48%) , Klebsiella pneumonia (15.40%) , Pseudomonas aeruginosa (8.50%) , Acinetobacter baumannii (5.04%) and Stenotropho-monas maltophilia (3.41%) respectively. CRE amounted to 123 (6.68%) . Common gram-positive bacteria were Staphylococcus aureus (4.92%) , Staphylococcus hominis (4.88%) and Staphylococcus epidermidis (4.71%) respectively. Candida albicans were the main fungus which accounted for 5.43%. The rates of Escherichia coli and Klebsiella pneumonia resistant to carbapenems were 3.5%-6.1% and 5.0%-6.3% respectively. The rates of Pseudomonas aeruginosa resistant to tobramycin and amikacin were 3.2% and 3.3% respectively. The resistant rates of Acinetobacter baumannii towards tobramycin and cefoperazone/sulbactam were both 19.2%. The rates of Stenotrophomonas maltophilia resistant to minocycline and sulfamethoxazole were 3.5% and 9.3% respectively. The rates of Staphylococcus aureus, Enterococcus faecium and Enterococcus faecalis resistant wards vancomycin were 0, 6.4% and 1.4% respectively; also, the rates of them resistant to linezolid were 1.2%, 0 and 1.6% respectively; in addition, the rates of them resistant to teicoplanin were 2.8%, 14.3% and 8.0% respectively. Furthermore, MRSA accounted for 39.15% (83/212) . Conclusions: Pathogens were mainly gram-negative bacteria. CRE accounted for 6.68%. The rates of Escherichia coli and Klebsiella pneumonia resistant to carbapenems were lower compared with other antibacterial agents. The rates of gram-positive bacteria resistant to vancomycin, linezolid and teicoplanin were still low. MRSA accounted for 39.15%.

[Primary ciliary dyskinesia with HYDIN gene mutations in a child and literature review].

Objective: To review children's primary ciliary dyskinesia (PCD) in the pathogenesis, clinical manifestation, diagnosis and treatment. Method: To summarize and analyze the clinical data of a patient who was admitted to the first affiliated hospital of Xiamen University with primary ciliary dyskinesia in April 2014 while referring to related literature. Result: An 11 years old boy, weighting about 22 kg, had a course of more than 10 years with repeated cough, stuffy and runny nose shortly after the birth. Examinations after admission to hospital showed that he presented with visible clubbing, bilateral paranasal sinus area tenderness, pharynx posterior wall with visible yellow pussy stuff drip and bilateral lung had scattered wet rales. Auxiliary examination revealed bilateral maxillary sinus, ethmoid sinus inflammation and bronchitis with left lower lung bronchiectasis. Fiberoptic bronchoscopy discovered congestion and a lot of sputum; ciliary biopsy pathology displayed that cilia were sparse and partial cilia 9+ 2 microtubules structural abnormalities. Full sequence of exon gene sequencing revealed two mutations located at chromosome 16 chr16: 71061369 (non-coding regions) and chr16: 70993591 (coding). Two novel mutations m. 3362A>G(E20) and c. 6101G>A(E39) in exon 16 of the HYDIN gene were identified. With the" ciliary motility disorder, gene" as keywords , the CNKI, Wanfang digital knowledge service platform and PubMed were searched for relevant articles from the establishment to July 2016. The studies retrieved included 9 cases and these cases were summarized. Comprehensive analysis showed that HYDIN gene mutations related PCD patients had the typical PCD performance such as repeatedly wet cough, sinusitis, bronchiectasis, and otitis media. The majority of patients have a history of acute respiratory distress syndrome in infancy and no visceral dislocation was not found. Most of the patients had no obvious structural abnormalities in cilia electron microscopic examination. Conclusion: The PCD patients with HYDIN genes mutations have clinical manifestations such as sinusitis, otitis media, bronchiectasis but without transposition of viscera. Cilia structure can be normal under the electron microscopic examination in some of patients.

[Application of the computer-based respiratory sound analysis system based on Mel-frequency cepstral coefficient and dynamic time warping in healthy children].

OBJECTIVE: We designed a computer-based respiratory sound analysis system to identify pediatric normal lung sound. To verify the validity of the computer-based respiratory sound analysis system. METHOD: First we downloaded the standard lung sounds from the network database (website: http: //www.easyauscultation.com/lung-sounds-reference-guide) and recorded 3 samples of abnormal loud sound (rhonchi, wheeze and crackles) from three patients of The Department of Pediatrics, the First Affiliated Hospital of Xiamen University. We regarded such lung sounds as"reference lung sounds". The"test lung sounds"were recorded from 29 children form Kindergarten of Xiamen University. we recorded lung sound by portable electronic stethoscope and valid lung sounds were selected by manual identification. We introduced Mel-frequency cepstral coefficient (MFCC) to extract lung sound features and dynamic time warping (DTW) for signal classification. RESULT: We had 39 standard lung sounds, recorded 58 test lung sounds. This computer-based respiratory sound analysis system was carried out in 58 lung sound recognition, correct identification of 52 times, error identification 6 times. Accuracy was 89.7%. CONCLUSION: Based on MFCC and DTW, our computer-based respiratory sound analysis system can effectively identify healthy lung sounds of children (accuracy can reach 89.7%), fully embodies the reliability of the lung sounds analysis system.

Bioequivalence of clopidogrel hydrogen sulfate tablets in healthy Chinese volunteers.

We aimed to evaluate the bioequivalence of clopidogrel in healthy Chinese volunteers after administration of a single oral dose. We administered a single oral dose of 75 mg clopidogrel (test and reference) to 32 healthy Chinese volunteers according to an open, randomized, crossover design. The concentration of clopidogrel acid (carboxylic metabolite of clopidogrel) in the plasma was determined using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Bioequivalence of the test and reference preparations were calculated using analysis of variance and one-sided t-test by using the DAS 2.0 software. The pharmacokinetic parameters of the test and reference preparations were as follows: peak plasma concentration (Cmax), 1351.101 ± 654.955 ng/mL and 1184.652 ± 607.713 ng/mL; area under the curve, 2642.017 ± 1093.848 ng·h/mL and 2780.666 ± 1283.100 ng·h/mL; and time to reach Cmax (Tmax), 0.789 ± 0.318 h and 0.953 ± 0.633 h, respectively. The relative bioavailability of the formulation was 101.7 ± 35.3%, which indicated that the test preparation was bioequivalent to the reference drug.

Characterization of a New Anastomosis Group (AG-W) of Binucleate Rhizoctonia, Causal Agent for Potato Stem Canker.

Two binucleate Rhizoctonia (BNR) isolates were recovered from potato cankered stems in Heilongjiang Province, northeastern China. Their cultural appearance on potato dextrose agar remained whitish as the cultures aged. White monilioid cells formed in the fluffy aerial hyphae, whereas no sclerotia appeared during the incubation. The two isolates could anastomose with each other, but they failed to anastomose with reference strains of BNR from AG-A to AG-Q, and AG-U. Analyses of restriction fragment length polymorphism (RFLP) of internal transcribed spacer of ribosomal DNA (rDNA-ITS) regions confirmed that these two isolates differed from the reference strains. The phylogenetic tree based on the sequences of rDNA-ITS regions showed that they were located in a distinct clade from other BNR AGs. These collective results suggested that the isolates recovered from potato in this study belonged to a new BNR AG designated as AG-W. Pathogenicity tests under glasshouse conditions revealed that both isolates were able to cause brown, dry, and slightly sunken lesions on potato subterranean stems. To our knowledge, this is the first report of the AG-W causing potato disease in China as well as worldwide.

Role of 1,25-dihydroxyvitamin D3 in the treatment of asthma.

OBJECTIVE: This study aims to observe the influence of 1,25-(OH)2D3 on airway inflammation and chemokine expression in asthmatic rats and to explore its significance in the treatment of asthma. MATERIALS AND METHODS: Wistar rats were randomly divided into a normal control group (N), an asthma group (A), a 1,25-(OH)2D3 group (VD), a budesonide group (P) and a 1,25-(OH)2D3 + budesonide treatment group (L). The acute asthma models were established through ovalbumin sensitisation and challenge. Lung tissues were stained with haematoxylin and eosin to observe pathologic changes, whereas an enzyme-linked immunosorbent assay was used to examine serum IgE, as well as the eosinophil chemoattractant protein (eotaxin) and interleukin-8 (IL-8) expression levels in bronchoalveolar lavage fluid and the serum. RESULTS: VD treatment partially reversed the characteristic pathological changes of airway inflammation. The IgE, eotaxin, and IL-8 expression levels in the VD group were significantly lower than those in the A group (p < 0.05) but remained higher than those in the control group (p < 0.05). CONCLUSIONS: 1,25-(OH)2D3 reduces airway inflammation, airway hyperresponsiveness and airway remodeling by partially inhibiting chemokine production during airway inflammation, and 1,25-(OH)2D3 synergises with hormone therapy.

First Report of Potato Stem Canker Caused by Binucleate Rhizoctonia AG-A in Jilin Province, China.

Potato (Solanum tuberosum L.) stem canker caused by Rhizoctonia solani occurs in potato-growing regions all over the world and can result in severe losses in crop yield and quality. In late July 2011, potato subterraneous stems with stem cankers composed of brownish, sunken lesions were observed at 15% incidence in seven sites in Jilin Province, northeast China. Samples were collected, and stem pieces (each 5 mm long) taken from the margins of the healthy and diseased tissues were surface-disinfested with 0.5% NaOCl for 2 min, rinsed with sterilized water, dried, then placed on potato dextrose agar at 25°C in the dark. Three (designated JL-3, JL-5-1, and JL-6) of seven Rhizoctonia isolates that developed from single hyphal tip transfers were identified preliminarily as binucleate Rhizoctonia (BNR) isolates (teleomorph Ceratobasidium Rogers). The colonies were white or light gray with fluffy aerial hyphae and no sclerotia after 14 days in culture. Hyphal cells were binucleate when stained with 4'-6-diamidino-2-phenylindole. Average hyphal diameters (mean ± standard deviation) of isolates JL-3, JL-5-1, and JL-6 were 4.8 ± 0.5 µm (range 4.1 to 5.6 µm), 4.4 ± 0.4 µm (range 3.9 to 5.2 µm), and 4.5 ± 0.3 µm (range 4.0 to 5.0 µm), respectively. The internal transcribed spacer (ITS) region of ribosomal DNA was amplified from genomic DNA with primers ITS1 and ITS4 and sequenced. BLASTn analysis indicated that the resulting sequences (GenBank Accession Nos. JX885459, JX885460, and JX885461 for JL-3, JL-5-1, and JL-6, respectively) were 100% identical to that of a Ceratobasidium sp. AG-A isolate CHR08-10 (HQ270171). So the three isolates were identified as BNR AG-A based on morphological and molecular characteristics. To determine pathogenicity of the BNR isolates, potato seed tubers (cv. Favorita), each with 3- to 5-mm-long sprouts, were inoculated with wheat seeds (sterilized by autoclaving twice at 121°C for 1 h with a 24-h interval between autoclavings) colonized with each isolate (1). One sprouted potato tuber was planted in a plastic pot with a single colonized wheat seed placed 10 mm above the uppermost sprout tip in a sand/sawdust mixture (1:2 v/v). Plants were incubated in a glasshouse at 25 to 27°C, and assessed after 21 days. The test was performed on 20 plants/isolate and the experiment was repeated. The incidence of plants inoculated with JL-3, JL-5-1, and JL-6 that developed stem canker symptoms averaged 11.1, 23.5, and 28.6%, respectively, whereas all control plants inoculated with sterilized wheat seeds remained asymptomatic. Rhizoctonia spp. were not reisolated from the control plants, whereas BNR isolates were reisolated consistently from symptomatic stems of the inoculated plants, and the identity confirmed by morphological and molecular characteristics as described above, fulfilling Koch's postulates. BNR AG-A has been reported to be pathogenic on soybean (Glycine max), pea (Pisum sativum), snap bean (Phaseolus vulgaris), and pak choi (Brassica chinensis) in China (4). Isolates of R. solani AG-3 are most often associated with potato stem canker (2), although unidentified BNR isolates were reported to cause mild symptoms on potato sprouts in Finland (1), and small lesions on potato roots and stems in the United Kingdom (3). To our knowledge, this is the first report of BNR AG-A causing potato stem canker in Jilin Province, one of the main potato-producing areas of China. References: (1) M. J. Lehtonen et al. Plant Pathol. 57:141, 2008. (2) L. Tsror. J. Phytopatology 158:649, 2010. (3) J. W. Woodhall et al. New Dis. Rep. 23:31, 2011. (4) G. H. Yang et al. J. Phytopathology 153:333, 2005.

First Report of Potato Stem Canker Caused by Rhizoctonia solani AG4 HGII in Gansu Province, China.

Black scurf and stem canker on potato (Solanum tuberosum L.), caused by Rhizoctonia solani, is an important disease throughout the world. Isolates of R. solani AG3 are the principal cause of these diseases on potato (2). In August 2011, at the tuber bulking growth stage, symptoms typically associated stem canker, including dark brown stem lesions, were observed on 20% of potato plants collected from 23 locations (about 2,000 ha) in Gansu Province, northwest China. Stem pieces (each 5 mm long) taken from the margins of the healthy and diseased tissues were surface-disinfected with 0.5% NaOCl for 2 min, rinsed with sterilized water, dried, then placed on potato dextrose agar (PDA) at 25°C in the dark. Twenty-nine fungal isolates taken from single hyphal tips were identified as R. solani based on morphological traits, including mycelium branched at right angles with a septum near the branch and a slight constriction at the branch base. Hyphal cells were determined to be multinucleate (4 to 10 nuclei/cell) when stained with 4'-6-diamidino-2-phenylindole (DAPI). Anastomosis groups were determined by pairing with reference strains (kindly provided by N. Kondo, Hokkaido University, Japan), and three isolates (designed GS-15, GS-24, and GS-25) anastomosed with isolates of R. solani AG4. The internal transcribed spacer (ITS) region of rDNA was amplified from genomic DNA of each of the three isolates with primers ITS1 and ITS4. The resulting sequences (GenBank Accession Nos. JX843818, JX843819, and JX843820) were 100% identical to those of >10 R. solani AG4 HGII isolates (e.g., HQ629873.1; isolate ND13). Therefore, based on the anastomosis assay and molecular characteristics, the three isolates were identified as R. solani AG4 HGII. To determine pathogenicity of the AG4 HGII isolates, potato seed tubers (cv. Favorita) with 3 to 5 mm long sprouts were inoculated with wheat seeds (sterilized by autoclaving twice at 121°C for 1 h with a 24 h interval between autoclavings) colonized with each isolate (1). One sprouted tuber was planted in a sterilized plastic pot (1 liter) with a single colonized wheat seed placed 10 mm above the uppermost sprout tip in a sand/sawdust mixture (1:2 v/v, with dry heat sterilization at 161°C for 4 h before use). Plants were incubated in a glasshouse maintained at 25 to 27°C. The test was performed on 20 plants for each isolate, and the experiment was repeated. After 3 weeks, control plants inoculated with sterilized wheat seeds remained asymptomatic, and no Rhizoctonia spp. were isolated from these plants, whereas all inoculated plants showed symptoms of stem canker. R. solani AG4 HGII was reisolated consistently from symptomatic stems, and the identity of the reisolates confirmed by the morphological and molecular characteristics mentioned above, fulfilling Koch's postulates. Potato stem canker caused by R. solani AG4 HGII was reported previously in the United States (3). To our knowledge, this is the first report of R. solani AG4 HGII causing stem canker on potato in Gansu Province, the main potato-producing area of China. R. solani AG4 HGII can cause sheath blight on corn in China (4), which is commonly grown in rotation with potato. This rotation could increase the risk of soilborne infection to either crop by R. solani AG4 HGII. References: (1) M. J. Lehtonen et al. Plant Pathol. 57:141, 2008. (2) L. Tsror. J. Phytopathol. 158:649, 2010. (3) J. W. Woodhall et al. Plant Dis. 96:1701, 2012. (4) X. Zhou et al. J. Shenyang Agric. Univ. 43:33, 2012.

Insulin with chondroitinase ABC treats the rat model of acute spinal cord injury.

Traumatic brain injury is often associated with acute spinal cord injury (ASCI). Insulin and chondroitinase ABC (ChABC) are both therapeutically effective, but the combined therapeutic effect of insulin and ChABC is still not clear. A combination of insulin and ChABC were used to treat a rat model of ASCI. This combination therapy prevented neuronal cell death by improving motor function, increasing cell growth and inhibiting cell apoptosis in ASCI rats. Expression of growth-associated protein 43, a marker of axonal re-growth, increased after combined treatment with insulin and ChABC. These results may provide a basis for a future method of treating ASCI.

Induction of human T-cell tolerance to pig xenoantigens via thymus transplantation in mice with an established human immune system.

Thymus xenotransplantation has been shown to induce tolerance to porcine xenografts in mice and to permit survival of alpha1,3Gal-transferase knockout porcine kidney xenografts for months in nonhuman primates. We evaluated the ability of porcine thymus xenotransplantation to induce human T-cell tolerance using a humanized mouse (hu-mouse) model, where a human immune system is preestablished by implantation of fetal human thymus tissue under the kidney capsule and intravenous injection of CD34(+) hematopoietic stem/progenitor cells. Human T-cell depletion with an anti-CD2 mAb following surgical removal of human thymic grafts prevented the initial rejection of porcine thymic xenografts in hu-mice. In these hu-mice, porcine thymic grafts were capable of supporting human thymopoiesis and T-cell development, and inducing human T-cell tolerance to porcine xenoantigens. Human T cells from these mice responded strongly to third-party pig, but not to the thymic donor swine leukocyte antigen (SLA)-matched pig stimulators in a mixed lymphocyte reaction (MLR) assay. Anti-pig xenoreactive antibodies declined in these hu-mice, whereas antibody levels increased in nontolerant animals that rejected porcine thymus grafts. These data show that porcine thymic xenotransplantation can induce donor-specific tolerance in immunocompetent hu-mice, supporting this approach for tolerance induction in clinical xenotransplantation.

Poly(ADP-ribose) polymerase-1 plays a role in suppressing mammary tumourigenesis in mice.

The DNA strand break-binding molecule, poly(ADP-ribose) polymerase-1 (PARP-1), plays a role in DNA repair, chromosomal stability, transcription and cell death. Accumulating evidence suggests that dysfunction of PARP-1 contributes to tumorigenesis. Here, we report that PARP-1 deficiency causes mammary carcinoma formation in female mice, and that the introduction of Trp53 mutations accelerates the onset and shortens the latency of mammary tumorigenesis. We show that PARP-1 deficiency results in chromosomal aneuploidy and centrosome amplification, which are substantiated by the inactivation of Trp53 in primary mammary epithelial (PME) cells. In addition, PARP-1 deficiency compromises p53 activation and impairs BRCA1 recruitment to the sites of DNA damage in PME cells. PARP-1 complementation partly rescues the defective DNA damage response mediated by p53 and BRCA1. The present study thus identifies a role of PARP-1 in suppressing mammary tumorigenesis in vivo and suggests that dysfunction of PARP-1 may be a risk factor for breast cancer in humans.

Tolerization of Gal alpha 1,3Gal-reactive B cells in pre-sensitized alpha 1,3-galactosyltransferase-deficient mice by nonmyeloablative induction of mixed chimerism.

Using a alpha 1,3-galactosyltransferase wild-type (GalT(+/+)) to deficient (GalT(-/-)) mouse bone marrow transplantation model, we have previously demonstrated that a non-myeloablative conditioning regimen is capable of permitting induction of allogeneic and xenogeneic mixed chimerism. Chimerism is associated with the rapid and lasting tolerization of anti-Gal alpha 1,3Gal (Gal) natural antibody (Ab)-producing B cells. However, one limitation of this model is that anti-Gal natural Ab levels are lower in GalT(-/-) mice than in humans and other primates. To overcome this limitation, we have now investigated the possibility of inducing such tolerance in GalT(-/-) mice that produce much higher levels of anti-Gal Abs due to presensitization with Gal-bearing xenogeneic cells. B6 GalT(-/-) mice that were pre-sensitized with rabbit red blood cells received non-myeloablative conditioning with depleting anti-CD4 and CD8 mAbs, 3Gy whole body and 7Gy thymic irradiation, and infusion of BALB/c GalT(+/+) bone marrow cells (BMC). Although engraftment of standard marrow doses was inhibited by the presensitization, long-lasting mixed chimerism could be induced in recipients of a high dose [160 x 10(6)] of allogeneic wild-type BMC. Achievement of persistent chimerism was associated with high levels of anti-Gal IgG(1) pretransplant, suggesting an inhibitory effect of non-complement-fixing IgG(1) Ab on anti-Gal-mediated marrow rejection. Induction of mixed chimerism was associated with a rapid disappearance of serum anti-Gal and tolerization of anti-Gal Ab-producing cells. B cells with anti-Gal receptors became undetectable in mixed chimeras. Mixed chimeras accepted subsequently transplanted donor-type GalT(+/+) hearts (> 140 days), whereas rapid (within 2 days) rejection of GalT(+/+) hearts occurred in conditioned control GalT(-/-) mice. In conclusion, when a high dose of GalT(+/+) BMC was administered to pre-sensitized GalT(-/-) mice, chimerism and tolerance were achieved. The absence of B cells with receptors recognizing Gal in mixed chimeras suggests a role for clonal deletion/receptor editing in the maintenance of B cell tolerance.

T cell and B cell tolerance to GALalpha1,3GAL-expressing heart xenografts is achieved in alpha1,3-galactosyltransferase-deficient mice by nonmyeloablative induction of mixed chimerism.

BACKGROUND: We have previously demonstrated that mixed xenogeneic chimerism and donor-specific T-cell tolerance can be induced in the rat-to-mouse species combination by using a relatively nontoxic, nonmyeloablative conditioning regimen. However, natural antibodies (NAbs) against Galalpha1,3Gal (Gal) pose an additional major barrier to pig-to-human vascularized xenograft acceptance. METHODS: To determine whether the mixed chimerism approach could also overcome this humoral barrier, T cell-depleted rat (GalT+/+) bone marrow cells (BMC) were transplanted to alpha1,3-galactosyltransferase deficient (GalT-/-) mice conditioned with a nonmyeloablative regimen, consisting of transient T cell and natural killer (NK) cell depletion, 3 Gy whole body irradiation, and 7 Gy thymic irradiation. RESULTS: By giving a high dose (180x106) of rat BMC, persistent mixed chimerism could be induced in GalT-/- mice, although the level of donor-type hematopoietic repopulation declined over time. Induction of mixed chimerism was associated with a rapid disappearance of anti-Gal and anti-rat NAb in the sera. Both anti-Gal Ab-producing cells and B cells with receptors recognizing Gal were undetectable in mixed chimeras, even when the chimerism levels declined, suggesting that a very low level of chimerism could effectively maintain B-cell tolerance to Gal, probably by clonal deletion and/or receptor editing. Mixed chimeras accepted subsequently transplanted donor-type rat hearts (>100 days) without immunosuppressive therapy, whereas delayed vascular and even hyperacute rejection of rat hearts occurred in conditioned control GalT-/- mice. Cellular rejection occurred by 5-6 days in conditioned control wild-type mice. CONCLUSIONS: These findings demonstrate that induction of mixed chimerism with a nonmyeloablative regimen can prevent vascularized xenograft rejection by cellular and anti-Gal Ab-dependent pathways in GalT+/+-to-GalT-/- species combinations.

[The bottleneck steps limiting maturation of penicillin G acylase in Escherichia coli].

We have identified the bottleneck steps limiting maturation of penicillin G acylase (PAC) through comparison of the maturation performance for various PAC-expression systems (Pac, Tac, T7, Vgb + T7) with different efficiencies of proteolysis, subunit folding and assembly. The maturation of PAC could be limited by various steps, such as translocation, periplasmic proteolysis, subunit folding and assembly depending on the host/vector systems. In BL21(pPA6) cells, maturation of PAC were limited by proteolysis and folding steps; the efficiency of proteolysis was 57.2%; the subunit folding and assembly capacity was 0.72. In BL21(pKKpacSP) cells, the stability and folding of alpha subunit was bottleneck steps. In T7 and dissolved-oxygen regulation expression systems, PAC proprecursor could be maturated efficiently. Results also indicate that the folding of alpha peptide plays a key role in folding of precursor for PAC in E. coli. Developing proper host/vector systems and fermentation technology with superior abilities on subunit folding and assembly of precursor for PAC could be plausible for enhancing production of PAC. In this study, pac could be expressed (transcribed, translated and maturated) efficiently under the control of T7 promoter.