[Repair methods of complex facial defect wounds involving paranasal sinuses and their clinical effectiveness].

Objective: To explore the repair methods of complex facial defect wounds involving paranasal sinuses and their clinical effectiveness. Methods: A retrospective observational study was conducted. From January 2020 to May 2022, 5 patients admitted to the Department of Burns and Plastic Surgery of Xiangya Hospital of Central South University and 4 patients admitted to the Department of Burns and Plastic Surgery of Chenzhou First People's Hospital with complex facial defect wounds involving paranasal sinuses met the inclusion criteria, including 6 males and 3 females, aged 35-69 years, including 4 patients with titanium mesh exposure combined with paranasal sinuses injury and 5 patients with tumor involving paranasal sinuses. After an adequate assessment of the damage by a multiple discipline team, titanium mesh removal, paranasal sinus debridement, and paranasal sinus mucosa removal were performed in patients with exposed titanium mesh, and radical tumor resection was performed in patients with tumors, with postoperative skin and soft tissue defects areas of 5.0 cm×2.5 cm to 18.0 cm×7.0 cm, anterior paranasal sinus wall defects/absence areas of 3 cm×2 cm to 6 cm×4 cm, and sinus cavity depths of 1 to 4 cm. Depending on the perforator course of the descending branch of the lateral circumflex femoral artery, the anterolateral femoral chimeric flap or anterolateral femoral myocutaneous flap (with flap area of 9 cm×4 cm to 19 cm×8 cm, muscle size of 5 cm×3 cm×3 cm to 11 cm×6 cm×3 cm) was transplanted to repair the defect, and the donor site wound was sutured directly. The type of tissue flap transplanted, the blood vessel of the recipient area, and the vascular anastomosis way during the operation, the recovery of the donor and recipient areas and the occurrence of complications after operation were observed. The appearance and blood supply of the recipient area and the recurrence of ulcers and tumors were followed up. Results: The anterolateral femoral myocutaneous flap transplantation was performed in 6 patients, and the anterolateral femoral chimeric flap transplantation was performed in 3 patients. The blood vessels in recipient areas were facial arteries and veins in 3 cases and superficial temporal arteries and veins in 6 cases. The superficial temporal arteries and veins were bridged with blood vessels in tissue flaps by flow-through way in 2 patients, and end-to-end anastomosis of blood vessels in donor and recipient areas was performed in 7 patients. After operation, all the tissue flaps survived, and the facial defect wounds were well repaired without cerebrospinal fluid leakage or paranasal sinus secretion leakage, no intracranial infection occurred, and the wounds in donor areas were healed well. Follow-up of 6-35 months after operation showed that all the patients had good blood supply in the recipient area, and the shape was acceptable; 4 patients with exposed titanium mesh had no recurrence of ulceration, and 5 patients with tumor had no local tumor recurrence or metastasis. Conclusions: Based on an adequate assessment of the extent of paranasal sinuses involved in the facial wound and the nature of the defect, good clinical effects can be achieved by using the anterolateral femoral muscle flap or the anterolateral femoral chimeric flap transplantation to repair complex facial defect wounds with open paranasal sinuses.

[Effect of CKIP-1 on hepatocyte apoptosis in nonalcoholic fatty liver disease].

Objective: To explore the effect and underlying mechanism of casein kinase 2 interacting protein-1 (CKIP-1) on hepatocyte apoptosis in nonalcoholic fatty liver disease (NAFLD). Methods: Experimental study. An NAFLD cell model was established by inducing human hepatoma cell line, HepG2 cells, with oleic acid (OA). Flag-CKIP-1 expression vector and shRNA-CKIP-1 were transfected into HepG2 cells. Flow cytometry was used to detect the effect of CKIP-1 on the activity and apoptosis of NAFLD hepatocytes. The levels of apoptosis-related proteins were detected by Western blot. CKIP-1 knockout mice in C57BL/6 back-ground were fed with either standard or high-fat diet for 8 weeks. Apoptosis-related signal proteins in NAFLD hepatocytes were detected by immunohistochemistry. Results: After CKIP-1 was transfected into HepG2 cells, the degree of OA induced cell liposis was significantly reduced (P<0.05). Annexin V-FITC/PI flow cytometry showed that CKIP-1 reduced the apoptosis of steatotic hepatocytes. Overexpression of CKIP-1 could significantly inhibit the expression of caspase-3 and caspase-9 and increase the expression of Bcl-2/Bax (P<0.05). Knockdown of CKIP-1 could increase the expression of caspase-3 and caspase-9 (P<0.05). CKIP-1 knockout could further increase the expression of caspase-3 and caspase-9 in NAFLD mice (P<0.01,P<0.05), and further decrease the expression of Bcl-2/Bax (P<0.05). Conclusion: CKIP-1 inhibited the apoptosis of steatotic hepatocytes by up-regulating the expression of apoptosis inhibitor gene, Bcl-2/Bax, and affecting the proteases, caspase-3 and caspase-9.

Construction of multifunctional porcine acellular dermal matrix hydrogel blended with vancomycin for hemorrhage control, antibacterial action, and tissue repair in infected trauma wounds.

Prevention of bacterial infection and reduction of hemorrhage, the primary challenges posed by trauma before hospitalization, are essential steps in prolonging the patient's life until they have been transported to a trauma center. Extracellular matrix (ECM) hydrogel is a promising biocompatible material for accelerating wound closure. However, due to the lack of antibacterial properties, this hydrogel is difficult to be applied to acute contaminated wounds. This study formulates an injectable dermal extracellular matrix hydrogel (porcine acellular dermal matrix (ADM)) as a scaffold for skin defect repair. The hydrogel combines vancomycin, an antimicrobial agent for inducing hemostasis, expediting antimicrobial activity, and promoting tissue repair. The hydrogel possesses a porous structure beneficial for the adsorption of vancomycin. The antimicrobial agent can be timely released from the hydrogel within an hour, which is less than the time taken by bacteria to infest an injury, with a cumulative release rate of approximately 80%, and thus enables a relatively fast bactericidal effect. The cytotoxicity investigation demonstrates the biocompatibility of the ADM hydrogel. Dynamic coagulation experiments reveal accelerated blood coagulation by the hydrogel. In vivo antibacterial and hemostatic experiments on a rat model indicate the healing of infected tissue and effective control of hemorrhaging by the hydrogel. Therefore, the vancomycin-loaded ADM hydrogel will be a viable biomaterial for controlling hemorrhage and preventing bacterial infections in trauma patients.

Study of charged particle activation analysis (II): Determination of boron concentration in human blood samples.

Boron Neutron Capture Therapy (BNCT) is a radiotherapy for the treatment of intractable cancer. In BNCT precise determination of 10B concentration in whole blood sample before neutron irradiation of the patient, as well as accurate neutron dosimetry, is crucial for control of the neutron irradiation time. For this purpose ICP-AES and neutron induced prompt γ-ray analysis are generally used. In Ibaraki Neutron Medical Research Center (iNMRC), an intense proton beam will be accelerated up to 8 MeV, which can also be used for Charged Particle Activation Analysis (CPAA). Thus, in this study, we apply the CPAA utilizing the proton beam to non-destructive and accurate determination of 10B concentration in whole blood sample. A CPAA experiment is performed by utilizing an 8 MeV proton beam from the tandem accelerator of Nuclear Science Research Institute in Japan Atomic Energy Agency. The 478 keV γ-ray of 7Be produced by the 10B(p, α)7Be reaction is used to quantify the 10B in human blood. The 478 keV γ-ray intensity is normalized by the intensities of the 847 keV and 1238 keV γ-rays of 56Co originating from Fe in blood. The normalization methods were found to be linear in the range of 3.27 µg 10B/g to 322 µg 10B/g with correlation coefficients of better than 0.9999.

[Research progress on Andersson lesion in ankylosing spondylitis].

Andersson lesion is a destructive vertebral or disco-vertebral lesion that occurs in the late stage of the ankylosing spondylitis. According to the etiology, these lesions are usually classified into localized lesions and extensive lesions. The history of ankylosing spondylitis and characteristic imaging is the key to the diagnosis of Andersson lesion. Conservative treatment may be effective for localized lesions. However, surgical intervention is often required for the failure of conservative treatment and extensive lesions. Currently, the optimal procedure for this problem is spinal osteotomy through pseudarthrosis and debridement via posterior-only approach.

Effectiveness of amylmetacresol and 2,4-dichlorobenzyl alcohol throat lozenges in patients with acute sore throat due to upper respiratory tract infection: a systematic review protocol.

REVIEW QUESTION/OBJECTIVE: This review aims to determine the best available evidence related to the effectiveness of amylmetacresol and 2,4-dichlorobenzyl alcohol throat lozenges in patients with acute sore throat due to upper respiratory tract infection (URTI). The objective is to examine the analgesic properties of amylmetacresol and 2,4-dichlorobenzyl alcohol (AMC/DCBA) throat lozenge comparing with placebo for the relief of pain in patients with acute sore throat due to URTIs.The review question is:More specifically, the objectives are to.

The pattern recognition receptor, Mincle, is essential for maintaining the M1 macrophage phenotype in acute renal inflammation.

Mincle (macrophage-inducible C-type lectin, Clec4e) is a transmembrane pattern recognition receptor involving the innate immunity, but its role in kidney disease is still unexplored. In the obstructed kidney of the unilateral ureteral obstruction model of renal injury, Mincle was specifically detected in the infiltrating M1 macrophages (CD68+iNOS+ cells) on day one but was rapidly reduced following reduction of M1 macrophages during the progression of kidney injury. Interestingly, Mincle-expressing macrophages were progressively increased in the cisplatin-induced acute kidney injury model, where iNOS expression was detected in the CD68+ cells following Mincle induction. Adaptive transfer of Mincle+ M1 macrophages largely promoted cisplatin-induced renal inflammation, which was prevented by the knockdown of Mincle in the transferred cells. Mincle was tightly regulated by TLR4/NF-κB signaling as evidenced by the binding of NF-κB/p65 to the promoter region of Mincle in LPS-primed macrophages. Blocking TLR4 or NF-κB suppressed LPS-induced Mincle expression on macrophages. Importantly, Mincle was found to be essential for maintaining the inflammatory phenotypes of M1 macrophages through the Syk signaling pathway since knockdown of Mincle or inhibition of Syk suppressed LPS-induced IL-1ß, MCP-1, and iNOS expression. Thus, Mincle is induced specifically on M1 macrophages, where Mincle-Syk signaling promotes and maintains inflammatory phenotypes of M1 macrophages in acute renal inflammation. Hence, targeting Mincle may be a novel therapy for acute kidney injury associated with M1 macrophages.

[Characteristics of sympathetic skin response in patients with multiple system atrophy].

Objective: To provide evidence for early clinical diagnosis of multiple system atrophy(MSA)by studying the characteristics of sympathetic skin responses(SSR) in the patients with MSA. Methods: A total of 47 MSA patients and 32 healthy individuals were enrolled as case group and normal control(NC) group, from in and out patients of Neurology Department of Navy General Hospital from July 2013 to August 2015. SSR was tested by Nicolet electromyography, the latency and abnormal and disappeared rate of SSR were compared. Results: The SSR latency of upper limbs and lower limbs in MSA group had statistical significance compared respectively with the NCgroup (upper limbs: SSR latency was(1 485±187)ms in MSA group, and(1 375±108)ms in NC group, P<0.05; lower limbs: SSR latency was(2 200±386)ms in MSA group, and(1 994±240)ms in NC group, P<0.05). Sex and age had no significant effect on the latency and the abnormal and disappeared rate of SSR in two groups (P>0.05). The upper and lower limb SSR latency in MSA patients with disease duration more than 2 years(SSR latency was (1 592±160)ms in upper limb and (2 268±254)ms in lower limb) were longer than those within 2 years(SSR latency was (1 453±184)ms in upper limb and (2 190±442)ms in lower limb), but only the upper limbs had significantly statistical differences (P<0.05). Both SSR abnormal rate and SSR disappeared rate in MSA patients whose disease duration were more than 2 years(SSR abnormal rate: 85.00%, SSR disappeared rate: 75.00%) were higher than those with shorter disease duration(SSR abnormal rate: 55.56%, SSR disappeared rate: 22.22%), and both were statistically significant (SSR abnormal rate: P<0.05, SSR disappeared rate: P<0.001). The upper and lower limb SSR latency of MSA-C subgroup had no statistical difference compared with MSA-P subgroup(P>0.05). The SSR abnormal rate in MSA-C subgroup(78.13%) was higher than that of MSA-P subgroup(46.76%), and were statistically significant (P<0.05). The SSR disappeared rate in MSA-C subgroup has no statistical difference compared with the MSA-P subgroup(P>0.05). Conclusions: SSR is helpful to diagnose MSA. The latency and the abnormal and disappeared rate of SSR are significantly increased with the extension of MSA duration. The SSR abnormal rate in MSA-C patients is higher than that in MSA-P patients, and symmetrically abnormal SSR is more supporting the diagnosis of MSA.

[The correlation factor analysis for conversion of clinically isolated syndrome to multiple sclerosis and neuromyelitis optica].

OBJECTIVE: To analyze the features of patients who converted from clinically isolated syndrome (CIS) to multiple sclerosis (MS) and neuromyelitis optica (NMO) and explore the correlated factors. METHODS: A total of 151 patients admitted in our unit as CIS from January 2009 to December 2014 were enrolled in the study. All patients were divided into the following four groups by locations of the initial lesion, which were the spinal cord, the optic nerve, the brain stem and the multifocal lesions. Data were collected at the baseline including demographics, expanded disability status scale (EDSS) score, site of CIS, presence or absence of cerebrospinal fluid (CSF) oligoclonal bands (OB) and serum aquaporin-4 antibody (AQP4-Ab), evoked potential (EP) and MRI lesions. The conversion rates from CIS to clinically definite MS or NMO were calculated and the correlated factors were explored. RESULTS: With a mean follow-up period of (44.11±17.62)months, 46/151(30.5%) patients converted to MS, 28/151 (18.5%) to definite NMO and 66/151 patients(43.7%)remained as CIS. Other patients were converted to optic neuritis(4/151), one-time transverse myelitis(3/151), acute disseminated encephalomyelitis (1/151) and Balo concentric sclerosis(3/151) . The EDSS score was significantly higher in patients converted to NMO than those converted to MS (P=0.003). The initial manifestation of optic neuritis significantly correlated with the conversion to NMO (P=0.000), while the initial manifestation of CIS with multifocal lesions significantly correlated with the conversion to MS (P=0.000). Neither the isolated BAEP (P=0.703), VEP (P=0.076), SEP (P=0.915) nor the combination of two (P=0.546)or three (P=1.000) of the above parameters could help to distinguish the conversion to MS or NMO. More patients with positive CSF-OB converted to MS (P=0.001), while more patients with positive serum AQP4-Ab converted to NMO (P=0.001). More patients were serum AQP4-Ab positive in those converted to NMO than those converted to MS (P=0.000). Lesions longer than three vertebral segments were dominant in patients converted to NMO (P=0.000). The logistic regression analysis revealed that factors correlated with conversion from CIS to MS were the initial CIS manifestation of multifocal lesions (OR=4.775, P=0.002), positive CSF-OB (OR=7.794, P=0.002) and VEP abnormality (OR=7.251, P=0.001). Factors correlated with conversion from CIS to NMO were female in gender (OR=12.536, P=0.019), positive serum AQP4-Ab (OR=36.410, P=0.002), lesions longer than three vertebral segments (OR=93.602, P=0.001), abnormal VEP and SEP (OR=18.448, P=0.002; OR=12.731, P=0.016). CONCLUSIONS: Factors correlated with the conversion from CIS to MS are initial CIS manifestation of multifocal lesions, positive CSF-OB and abnormal VEP, while those correlated with the conversion from CIS to NMO are female in gender, positive serum AQP4-Ab, initial CIS manifestation with optic nerve, lesions involved more than three vertebral segments and abnormal VEP and SEP.

[Analysis of the survival of 123 patients with intrahepatic cholangiocarcinoma after surgical resection].

OBJECTIVE: To investigate the prognostic factors for patients with intrahepatic cholangiocarcinoma after surgical resection. METHODS: The clinicopathological and follow-up data of 123 patients with intrahepatic cholangiocarcinoma who underwent surgical resection in Cancer Hospital, Chinese Academy of Medical Sciences between January 1999 and June 2015 were collected and reviewed, and their survival and prognosis were analyzed with the Kaplan-Meier method and Cox regression model. RESULTS: The median follow-up time was 22 months and median recurrence-free survival time was 8.97 months. The 1-, 2- and 3-year recurrence rates were 58.6%, 68.9% and 76.5%, respectively. Cox multivariate regression analysis showed that preoperative rise in CEA, lymph node metastasis, multiple lesions, extrahepatic invasion, and combination of tumor necrosis were significant adverse prognostic factors affecting the postoperative recurrence-free survival in patients with intrahepatic cholangiocarcinoma after surgical resection (P<0.05 for all). The median overall survival time was 21.17 months, and the 1-, 3- and 5-year overall survival rates were 76.6%, 33.2% and 26.1%, respectively. The Cox multivariate regression analysis showed that preoperative rise in CEA, lymph node metastasis, multiple lesions, and extrahepatic invasion were significant adverse prognostic factors affecting the postoperative overall survival in patients with intrahepatic cholangiocarcinoma after surgical resection(P<0.05 for all). CONCLUSION: Preoperative rise in CEA, lymph node metastasis, multiple lesions and extrahepatic invasion are significant adverse prognostic factors for patients with intrahepatic cholangiocarcinoma after surgical resection.

Lipid disorder and intrahepatic renin-angiotensin system activation synergistically contribute to non-alcoholic fatty liver disease.

BACKGROUND: This study aimed to investigate the possible synergistic effects of lipid disorder with renin-angiotensin system (RAS) activation in non-alcoholic fatty liver disease (NAFLD). METHODS: Apolipoprotein E gene-knockout mice, angiotensin II (Ang II) type 1 receptor (AT1) gene-knockout mice and human hepatoblastoma cell line (HepG2) were used for experiments. Lipid accumulation was examined by Filipin staining and intracellular cholesterol quantitative assay. The gene and protein expression of molecules involved in RAS and low-density lipoprotein receptor (LDLr) pathway was examined by real-time PCR, immunofluorescent staining and Western blot. RESULTS: There was significantly increased expression of RAS components and extracellular matrix (ECM) in livers of high-fat-diet-fed apolipoprotein E gene-knockout mice compared with controls. Upregulation of RAS components was positively associated with increased plasma levels of lipid profile. The in vitro study further confirmed that cholesterol loading increased supernatant renin activity and Ang II level of HepG2 cells, accompanied by increased ECM production that was positively associated with increased expression of intracellular RAS components. Interestingly, Ang II treatment increased lipid accumulation in livers of C57BL/6 mice and HepG2 cells. Furthermore, Ang II treatment increased gene and protein expression of sterol regulatory element-binding protein (SREBP) cleavage activating protein (SCAP), SREBP-2 and LDLr, which were mediated by enhanced SCAP/SREBP-2 complex translocation from endoplasmic reticulum to Golgi. However, LDLr pathway was accordingly downregulated in livers of AT1 gene-knockout C57BL/6 mice or in HepG2 cells treated by telmisartan. CONCLUSION: These findings demonstrate that lipid disorder and intrahepatic RAS activation synergistically accelerate NAFLD progression.

MMSET regulates expression of IRF4 in t(4;14) myeloma and its silencing potentiates the effect of bortezomib.

Multiple myeloma (MM) is characterized by recurrent chromosomal translocations. In t(4;14) MM, the MM SET domain (MMSET) protein is universally overexpressed and has been suggested to have an important tumorigenic role. However, the exact molecular targets underlying MMSET activity are not well understood. Here, we found in t(4;14) MM cells that MMSET knockdown decreases interferon regulatory factor 4 (IRF4) expression, and ectopic MMSET increases IRF4 expression, suggesting that MMSET is an upstream regulator of IRF4. Further analyses indicated an interaction between MMSET and nuclear factor-κB, which both bind to the IRF4 promoter region. A luciferase reporter assay showed that MMSET is an important functional element for the IRF4 promoter. MMSET knockdown induces apoptosis and potentiates the effects of bortezomib in vitro and in vivo. Importantly, we found that bortezomib could reduce expression of MMSET and IRF4. This might partly explain the additive effect of combining MMSET knockdown and bortezomib treatment. These results identify MMSET as a key regulator involved in the regulatory network of transcription factor IRF4, which is critical for MM cell survival, suggesting that the combination of MMSET inhibition and bortezomib is likely to improve patient outcome in MM.

[Influence of BMP-7 on chondrocyte secretion and expression of Col-II,AGG and Sox9 mRNA in porous tantalum-chondrocyte composites in vitro].

OBJECTIVE: To study the influence of bone morphogenetic protein-7 (BMP-7) on chondrocyte secretion and expression of type II collagen (Col-II), aggrecan (AGG) and SRY-related high mobility group-box gene 9 (Sox9) mRNA in porous tantalum-chondrocyte composites. METHODS: The articular chondrocytes were isolated from 3-week-old New Zealand immature rabbits and identified. The 2nd generation of chondrocytes with 1×10(6)/mL inoculate concentration was seeded in porous tantalum and divided into 4 groups, and control group (tantalum/chondrocyte), 50 µg/L BMP-7 group (50 µg/L BMP-7/tantalum/chondrocyte), 100 µg/L BMP-7 group (100 µg/L BMP-7/tantalum/chondrocyte), and 200 µg/L BMP-7 group (200 µg/L BMP-7/tantalum/chondrocyte). The proliferation of chondrocytes was measured by CCK-8 assay. The chondrocyte growth and morphology were observed by scanning electron microscopy (SEM). The synthesis of glycosaminoglycan (GAG) in chondrocytes was tested by dimethyl methylene blue (DMMB) colorimetric quantification method. Col-II, AGG and Sox9 mRNA in chondrocytes were detected by real-time PCR. RESULTS: The chondrocytes were spindle-shaped in 24 hours of primary cell culture and most cells became polygonal shaped in 4 days. The chondrocytes were affirmed by alcian blue, safranin O and Col-II immunocytochemistry staining. The result of CCK-8 assay showed that the level of cell proliferation in 100 µg/L BMP-7 groups were higher than those in the other groups (P<0.05). The chondrocytes implanted into porous tantalum scaffolds with BMP-7 had better functions, by which cytoplasmic processes developed and extended to the surface and inner of porous tantalum by SEM observation. DMMB quantitative determination of GAG showed that GAG amount of chondrocytes in 100 µg/L BMP-7 groups was significantly higher than those in the other groups (P<0.05). The expressions of Col-II, AGG and Sox9 mRNA in chondrocytes were up-regulated in the experimental groups, compared with the control group and the best effect appeared when concentration of BMP-7 was 200 µg/L. (P<0.05). CONCLUSION: BMP-7/tantalum/chondrocytes composites enhanced in vitro chondrocyte proliferation and extracellular matrix greatly, and can promote chondrogenic gene expression.

Prevalence of Mycoplasma pneumoniae: a cause for community-acquired infection among pediatric populaztion.

BACKGROUND: Atypical pneumonia caused by Mycoplasma pneumoniae is a leading cause of mortality among the pediatric age group. OBJECTIVES: Our study was designed to know the prevalence of M. pneumoniae in children with community-acquired pneumonia and the involvement in the cytoadherence to the respiratory epithelium by M. pneumoniae using electron microscopy and immuno-gold labeling technique. MATERIALS AND METHODS: A total of 152 children of 1 month to 12 years of age of both sexes attending Hebei Provincial People's Hospital, Shijiazhuang, Hebei with diagnosed pneumonia were included in the study. RESULTS: Out of 152 children 84 (55.3%) were males, and 68 (44.7%) were females. The mean age of the patients in the control group (50 patients) was 18.5 ± 3 months with 31 (62%) males and 19 (38%) females. IgM antibodies against M. pneumoniae were positive in 84 (55.3%) males and 68 (44.7%) females. Out of 50 patients 9 (18%) were found to positive for IgM M. pneumoniae antibodies of which four (44.4%) males and 5 (55.5%) females were positive. Our study observed that the gold particles were clustered on the filamentous extension of the tip of the cells. Out of 152 serum samples subjected to particle agglutination assay 138 (90.7%) were positive 1:320 titer, 9 were >1:80 and 3 showed titer was >1:40. CONCLUSION: We suggest that clinicians should consider empirical therapy of broad spectrum antibiotics therapy to cover these atypical pathogens to reduce the severity before obtaining the serological results. From our study, we also suggest electron microscopic and biochemical studies for better diagnosis of these pathogens.

Effect of siRNA targeting EZH2 on cell viability and apoptosis of bladder cancer T24 cells.

We investigated the effect of siRNA targeting enhancer of EZH2 on cell proliferation, invasion, migration, and apoptosis of human bladder cancer T24 cells. An siRNA-expressing plasmid targeting the EZH2 gene was transfected into T24 cells. Quantitative polymerase chain reaction and Western blot analysis were used to detect EZH2 expression at the mRNA and protein levels, respectively. Proliferation, invasion, and migration of T24 cells were examined in vivo using MTT, wound healing, and transwell chamber migration assays, respectively. Annexin V-fluorescein isothiocyanate/propidium iodide flow cytometric analysis was performed to determine cell apoptosis levels. Expression of EZH2 in T24 cells was suppressed at the mRNA and protein levels. Following transfection for 48 h, growth was inhibited by 37.9%, which was markedly lower than that in the negative control group (P < 0.05). Following a wound-healing assay for 24 h, transfected cell migration distance was 1.37 ± 0.12, which was markedly less than the horizontal migration distance of negative control group cells (P < 0.01). In addition, the cell invasion ability of EZH2- siRNA group cells decreased by 67% compared with negative control group cells (P < 0.01). Following transfection for 48 h, early- and late-stage apoptosis rates for T24 cells were 22.8 and 3.60%, respectively, which were higher than in the negative control group (P < 0.01). EZH2 gene silencing effectively suppressed the proliferation, invasion, and migration abilities of human bladder cancer cells, promoting apoptosis.

Specific ion effects on the electrophoretic mobility of small, highly charged peptides: a modeling study.

In this work, we use coarse-grained modeling to study the free solution electrophoretic mobility of small highly charged peptides (lysine, arginine, and short oligos thereof (up to nonapeptides)) in NaCl and Na2SO4 aqueous solutions at neutral pH and room temperature. The experimental data are taken from the literature. A bead modeling methodology that treats the electrostatics at the level of the nonlinear Poisson Boltzmann equation developed previously in our laboratory is able to account for the mobility of all peptides in NaCl, but not Na2SO4. The peptide mobilities in Na2SO4 can be accounted for by including sulfate binding in the model and this is proposed as one possible explanation for the discrepancy. Oligo arginine peptides bind more sulfate than oligo lysines and sulfate binding increases with the oligo length.

Bioweathering of nontronite colloids in hybrid silica gel: implications for iron mobilization.

AIMS: This study aimed to study biotic iron dissolution using a new hybrid material constituted of well-dispersed mineral colloids in a silica gel matrix. This permitted to prevent adsorption of colloidal mineral particles on bacteria. Hybrid silica gel (HSG) permitted to study bioweathering mechanisms by diffusing molecules. METHODS AND RESULTS: Hybrid silica gel was synthesized through a classical sol-gel procedure in which mineral colloidal particles (NAu-2) were embedded in a porous silica matrix. Rahnella aquatilis RA1, isolated from a wheat rhizosphere was chosen for its ability to dissolve minerals by producing various organic acids and siderophores. Pyruvic, acetic and lactic acids were the major organic acids produced by R. aquatilis RA1 followed by oxalic and citric acids at the end of incubation. Comparison of abiotic and biotic experiments revealed a high efficiency of R. aquatilis RA1 for iron dissolution suggesting an optimized action of different ligands that solubilized or mobilized iron. CONCLUSIONS: Hybrid silica gel allowed focusing on the colloidal mineral weathering by metabolites diffusion without mineral adsorption on bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: Hybrid silica gels are new and efficient tools to study colloidal mineral bioweathering. Adjusting HSG porosity and hydrophobicity should permit to precise the influence of limiting diffusion of siderophores or aliphatic organic acids on mineral weathering.

First Report of a Leaf Spot on Goldthread (Coptis chinensis) Caused by Phoma aquilegiicola in China.

Goldthread (Coptis chinensis) is an important herbaceous plant in traditional Chinese medicine (3). Annual production of goldthread root is ~3,000 tons (dry weight) in China. The plant is cultivated extensively in Shizhu Co., Chongqing (29.98°E, 108.13°N), where goldthread yields account for more than 60% of total world production. A foliar disease was first observed on goldthread plants in 2008 in Shizhu County (5). In 2011 and 2012, about 10 ha of goldthread fields in different townships of Shizhu Co. were surveyed. The results demonstrated that the disease present in the fields was widespread at incidences of 30 to 100%, with yield losses of 15 to 75%. Typical symptoms included irregular, purple brown lesions on leaves, beginning usually at the leaf margin and extending to the central leaf blade. The lesions coalesced and turned deep purple. Black pycnidia were visible on the lesions, and severely diseased plants were usually wholly blighted. To identify the pathogen, infected leaves were collected from goldthread fields in different townships of Shizhu Co. and small pieces of symptomatic tissue were cut from each leaf. The leaf pieces were surface-disinfected for 1 min in 1.5% sodium hypochlorite, rinsed in sterilized water, air-dried, and transferred onto potato dextrose agar (PDA) plates with 0.5 g/liter of streptomycin sulfate. Thirty-three fungal isolates with similar colony morphology were obtained. On oatmeal agar plates, each colony was circular with a smooth edge, initially cream, and then pale-brown. Pycnidia were dark brown, spherical, with or without papillae, and 100 to 112 × 189 to 222 µm. Conidia were produced on short, straight, and aseptate conidiophores in the pycnidia; they were monocellular, hyaline, ellipsoidal or clavate, and 2.01 to 2.50 × 4.20 to 5.55 µm. Three isolates (SZ-9, SZ-10, and SZ-13) were selected randomly from all 33 isolates, and genomic DNA of each isolate was extracted following the CTAB method (4). The rDNA ITS region of each isolate was amplified with V9G/ITS4 primers and sequenced (1). The ITS sequences of the three isolates (GenBank Accession Nos. KF692355.2 [SZ-9], KF985236.1 [SZ-10], and KF985237.1 [SZ-13]) were identical, and BLAST revealed 100% identity with the ITS sequence of an isolate of Phoma aquilegiicola (CBS 107.96, GU237735.1). Based on the morphological characteristics and ITS sequences, all three isolates were identified as P. aquilegiicola. Pathogenicity test of 10 isolates was conducted by placing a 5-mm-diameter mycelial agar plug (from the margin of a 5-day-old PDA culture) on each of 10 fully-expanded leaves of healthy goldthread plants/isolate. Ten leaves were treated similarly with sterilized PDA plugs as a control. Inoculated and control plants were incubated in the dark for 24 h at 25 ± 2°C and >90% RH, and then maintained in a growth chamber at 25 ± 2°C, 3,100 lux, and >90% RH. The pathogenicity test was carried out three times. Symptoms developed on all inoculated leaves for all 10 isolates, but not on the control plants. Lesions were first visible 48 h after inoculation, and typical irregular lesions similar to those observed on field plants were seen after 6 days. The same pathogenic fungus was re-isolated from the infected leaves but not from the non-inoculated leaves. A disease caused by P. aquilegiicola was first reported on Aquilegia flabellata plants of the cv. Fan Columbine in a perennial garden in Italy (2). This is the first report of leaf spot on goldthread caused by P. aquilegiicola in China. Studies on the epidemiology and control of the disease are necessary owing to the economic significance of the host and destructiveness of the disease. References: (1) M. M. Aveskamp et al. Mycologia 101:363, 2009. (2) A. Garibaldi et al. Plant Dis. 95:880, 2011. (3) B. Liu et al. J. Pharmaceut. Biomed. 41:1056, 2006. (4) M. A. Saghai-Maaroof et al. Proc. Natl. Acad. Sci. USA. 81:8014, 1984. (5) X. R. Zhou et al. J. Shizhen Medicine Res. 23:471, 2012.

Genetic diversity among red swamp crayfish (Procambarus clarkii) populations in the middle and lower reaches of the Yangtze River based on AFLP markers.

The red swamp crayfish has become one of the most important freshwater aquaculture species in China. At present, although it is widely distributed in the middle and lower reaches of the Yangze River basin, little is known about its population genetics and geographic distribution in China. We estimated the genetic diversity among 6 crayfish populations from 4 lakes (Hongze Lake, Poyang Lake, Dongting Lake, and Yue Lake) using AFLPs. A total of 129 loci were generated with 5 EcoRI-MseI primer combinations and scored as binary data in 139 individuals. These data were analyzed by cluster methods with the NTSYSpc software package. The 6 populations were separated into 3 major clusters by principal coordinate analysis and cluster analysis. Among the 6 populations, the highest gene diversity was found within the Nanjing population. Analysis of molecular variance demonstrated that most variation occurred within populations (91.20%). The estimated average GST value across all loci was 0.4186, suggesting (very) low gene flow among the different localities. We conclude that there is high genetic differentiation among crayfish in the middle and lower reaches of the Yangze River. This information will help in the selection of high-quality individuals for artificial reproduction.

The effect of vitamin E on growth performance and meat quality in broilers given diets containing distillers' dried grain with solubles (DDGS).

1. The objective of the present study was to investigate the effect of vitamin E (VE, α-tocopherol acetate) on growth performance and meat quality of broilers fed on diets containing maize distillers' dried grains with solubles (DDGS). 2. A total of 360 one-d-old broilers were randomly assigned to 6 groups. Each group comprised 6 replicates of 10 broilers. Broilers were allocated to 1 of 6 diets and fed for 49 d in a 3 × 2 factorial design. The dietary treatments included three concentrations of DDGS (0, 10 or 20%) and two concentrations of VE (0 or 200 mg/kg). 3. Diets containing 20% DDGS decreased the growth phase and average daily feed intake, demonstrating that small amounts of DDGS affected feed intake in broilers and that an excess of DDGS was not conducive to feed intake. 4. Different dietary concentrations of DDGS did not significantly improve colour, drip loss, cooking loss, or shear force. Supplementation with 200 mg/kg VE significantly reduced cooking loss and shear force and improved colour. 5. In conclusion, supplementation with DDGS at 10% and/or α-tocopherol acetate at 200 mg/kg had positive effects on growth performance and meat quality in broilers.