Purpose: The poor retention and ambiguous differentiation of stem cells (SCs) within corpus cavernosum (CC) limit the cell application in erectile dysfunction (ED). Herein, the effects and mechanism of microRNA-145 (miR-145) gene modification on modulating the traits and fate of bone marrow-derived mesenchymal stem cells (BMSCs) were investigated. Materials and Methods: The effects of miR-145 on cell apoptosis, proliferation, migration, and differentiation were determined by flow cytometry, cell counting kit-8, transwell assays and myogenic induction. Then, the age-related ED rats were recruited to four groups including phosphate buffer saline, BMSC, vector-BMSC, overexpressed-miR-145-BMSC groups. After cell transplantation, the CC were harvested and prepared to demonstrate the retention and differentiation of BMSCs by immunofluorescent staining. Then, the target of miR-145 was verified by quantitative real-time polymerase chain reaction and immunohistochemical. After that, APTO-253, as an inducer of Krüppel-like factor 4 (KLF4), was introduced for rescue experiments in corpus cavernosum smooth muscle cells (CCSMCs) under the co-culture system. Results: In vitro, miR-145 inhibited the migration and apoptosis of BMSCs and promoted the differentiation of BMSCs into smooth muscle-like cells with stronger contractility. In vivo, the amount of 5-ethynyl-2′-deoxyuridine (EdU)+cells within CC was significantly enhanced and maintained in the miR-145 gene modified BMSC group. The EdU/CD31 co-staning was detected, however, no co-staining of EdU/α-actin was observed. Furthermore, miR-145, which secreted from the gene modified BMSCs, dampened the expression of KLF4. However, the effects of miR-145 on CCSMCs could be rescued by APTO-253. Conclusions Overall, miR-145 modification prolongs the retention of the transplanted BMSCs within the CC, and this effect might be attributed to the modulation of the miR-145/KLF4 axis. Consequently, our findings offer a promising and innovative strategy to enhance the local stem cell-based treatments.

OBJECTIVE: To prepare a novel hyaluronic acid methacrylate (HAMA) hydrogel microspheres loaded polyhedral oligomeric silsesquioxane-diclofenac sodium (POSS-DS) patricles, then investigate its physicochemical characteristics and in vitro and in vivo biological properties. METHODS: Using sulfhydryl POSS (POSS-SH) as a nano-construction platform, polyethylene glycol and DS were chemically linked through the "click chemistry" method to construct functional nanoparticle POSS-DS. The composition was analyzed by nuclear magnetic resonance spectroscopy and the morphology was characterized by transmission electron microscopy. In order to achieve drug sustained release, POSS-DS was encapsulated in HAMA, and hybrid hydrogel microspheres were prepared by microfluidic technology, namely HAMA@POSS-DS. The morphology of the hybrid hydrogel microspheres was characterized by optical microscope and scanning electron microscope. The in vitro degradation and drug release efficiency were observed. Cell counting kit 8 (CCK-8) and live/dead staining were used to detect the effect on chondrocyte proliferation. Moreover, a chondrocyte inflammation model was constructed and cultured with HAMA@POSS-DS. The relevant inflammatory indicators, including collagen type Ⅱ, aggrecan (AGG), matrix metalloproteinase 13 (MMP-13), recombinant A disintegrin and metalloproteinase with thrombospondin 5 (Adamts5), and recombinant tachykinin precursor 1 (TAC1) were detected by immunofluorescence staining and real-time fluorescence quantitative PCR, with normal cultured chondrocytes and the chondrocyte inflammation model without treatment as control group and blank group respectively to further evaluate their anti-inflammatory activity. Finally, by constructing a rat model of knee osteoarthritis, the effectiveness of HAMA@POSS-DS on osteoarthritis was evaluated by X-ray film and Micro-CT examination. RESULTS: The overall particle size of POSS-DS nanoparticles was uniform with a diameter of about 100 nm. HAMA@POSS-DS hydrogel microspheres were opaque spheres with a diameter of about 100 μm and a spherical porous structure. The degradation period was 9 weeks, during which the loaded POSS-DS nanoparticles were slowly released. CCK-8 and live/dead staining showed no obvious cytotoxicity at HAMA@POSS-DS, and POSS-DS released by HAMA@POSS-DS significantly promoted cell proliferation (P<0.05). In the chondrocyte anti-inflammatory experiment, the relative expression of collagen type Ⅱ mRNA in HAMA@POSS-DS group was significantly higher than that in control group and blank group (P<0.05). The relative expression level of AGG mRNA was significantly higher than that of blank group (P<0.05). The relative expressions of MMP-13, Adamts5, and TAC1 mRNA in HAMA@POSS-DS group were significantly lower than those in blank group (P<0.05). In vivo experiments showed that the joint space width decreased after operation in rats with osteoarthritis, but HAMA@POSS-DS delayed the process of joint space narrowing and significantly improved the periarticular osteophytosis (P<0.05). CONCLUSION HAMA@POSS-DS can effectively regulate the local inflammatory microenvironment and significantly promote chondrocyte proliferation, which is conducive to promoting cartilage regeneration and repair in osteoarthritis.
Animals ; Rats ; Matrix Metalloproteinase 13 ; Microspheres ; Hydrogels ; Collagen Type II ; Diclofenac ; Inflammation ; Osteoarthritis, Knee/drug therapy* ; Hyaluronic Acid ; Aggrecans

Protein nitration and nitrosylation are essential post-translational modifications (PTMs) involved in many fundamental cellular processes. Recent studies have revealed that excessive levels of nitration and nitrosylation in some critical proteins are linked to numerous chronic diseases. Therefore, the identification of substrates that undergo such modifications in a site-specific manner is an important research topic in the community and will provide candidates for targeted therapy. In this study, we aimed to develop a computational tool for predicting nitration and nitrosylation sites in proteins. We first constructed four types of encoding features, including positional amino acid distributions, sequence contextual dependencies, physicochemical properties, and position-specific scoring features, to represent the modified residues. Based on these encoding features, we established a predictor called DeepNitro using deep learning methods for predicting protein nitration and nitrosylation. Using n-fold cross-validation, our evaluation shows great AUC values for DeepNitro, 0.65 for tyrosine nitration, 0.80 for tryptophan nitration, and 0.70 for cysteine nitrosylation, respectively, demonstrating the robustness and reliability of our tool. Also, when tested in the independent dataset, DeepNitro is substantially superior to other similar tools with a 7%-42% improvement in the prediction performance. Taken together, the application of deep learning method and novel encoding schemes, especially the position-specific scoring feature, greatly improves the accuracy of nitration and nitrosylation site prediction and may facilitate the prediction of other PTM sites. DeepNitro is implemented in JAVA and PHP and is freely available for academic research at http://deepnitro.renlab.org.
Amino Acid Sequence ; Amino Acids ; metabolism ; Deep Learning ; Humans ; Internet ; Neural Networks (Computer) ; Nitrosation ; Proteins ; chemistry ; metabolism ; Reproducibility of Results ; Software

Objective To evaluate how proprioception affect ankle stability through comparing angle position awareness and peroneus reaction time between chronic ankle lateral instability patients and healthy controls.Methods A total of 51 participants were recruited into an experimental group of 21 patients with chronic ankle lateral instability (17 males,aged 31.6±2.6) and a control group of 30 healthy counterparts (24 males,aged 34.2±2.3).All the participants were asked to reoccur passive ankle position under the angular velocity at 2 degree per second when they were resting with non-weight bearing in their recruited ankles.The muscle reaction time (MRT)of peroneus longus(PL) and peroneus brevis (PB) in all the recruited ankles was measured during sudden ankle inversion both with and without ankle protective brace wearing.Results The difference between angle recurrence and the target angle (ankle inversion 20° and 30°) was significantly higher (P＜0.05) in the experimental group compared to the control group.The average MRTs of PL and PB were also significantly longer (P＜0.05) in the experimental group than the control group,whether wearing ankle protective braces or not.However,within both groups,no significant differences of PL and PB's MRT were identified between brace wearing and no brace (P＞0.05).Conclusions In patients with chronic lateral ankle instability,the position awareness decreases and the reaction time of peroneus is prolonged.Ankle braces can provide mechanical protection to the injured joints,but cannot promote MRT significantly.

Objective To explore the diagnostic value of 18F-FDG PET/CT delayed imaging after induced diuresis for primary bladder tumor.Methods Fourteen patients (12 males,2 females; age range 35-88 years) with pathologically confirmed primary bladder tumor (clinical stage T1.3N0M0) were retrospectively analyzed.All patients underwent standard 18F-FDG PET/CT followed by a delayed (2.5-3.0 h later)pelvic imaging post intravenous injection of 20 mg furosemide and oral intake of 600 ml water.A positivelesion was defined as the uptake of 18F-FDG greater than the urine radioactivity and negative as equaled to or less than the urine radioactivity.Diagnostic efficacy of 18F-FDG PET/CT was calculated with pathology as the gold standard.Mann-Whitney u test was used to analyze data.Results There were 12 malignant (11 urothelium carcinomas and 1 squamous cell carcinoma) and 2 benign tumors (papillary epitheliomas).On standard PET/CT imaging,3 of 12 malignant lesions showed increased metabolism of 18F-FDG and the other 9 were false negative; while the 2 benign tumors showed no abnormal 18F-FDG uptake.The sensitivity,specificity,and accuracy of 18F-FDG PET/CT were 3/12,2/2 and 5/14,respectively.Sizes of the 3 18F-FDG-avid malignant lesions were greater than those of other 9 18F-FDG-negative lesions (26,30,35 mm vs (15.6±6.3) mm; Z=-2.315,P＜0.05).The sensitivity,specificity,and accuracy of delayed pelvic imaging were 11/12,2/2,13/14,respectively.Conclusion 18F-FDG PET/CT delayed imaging after diuresis has a high diagnostic efficacy for primary bladder tumor.

Objective To explore the diagnostic value of delayed 18 F?FDG PET/CT pelvic imaging after forced diuresis in detecting male patients with locally recurrent rectal cancer. Methods Thirty?two male patients ( 32-79 years) with clinically suspicious recurrences of rectal cancer were retrospectively ana?lyzed. All patients underwent a standard 18 F?FDG PET/CT followed by a delayed pelvic imaging after 2.5 h of administration of 20 mg furosemide intravenously and extra water intake of 500 ml at least. Final diagnosis was made by pathology or clinical follow?up (>10 months ) . Diagnostic efficiency of 18 F?FDG PET/CT standard and delayed pelvic images were calculated and compared. χ2 test was used for data analysis. Re?sults A total of 25 patients were diagnosed with recurrent rectal cancer, including 10 cases with invasion of surrounding tissue, and 5 cases with metastases. Among the 7 patients without local recurrences, 2 had dis?tant metastases. The sensitivity, specificity, and accuracy of standard FDG PET/CT were 92.0%(23/25), 3/7, 81.2%(26/32), respectively. The corresponding values of the delayed imaging were 96.0%(24/25), 6/7, and 93.8%(30/32), respectively. The specificity of delayed imaging was higher than that of standard FDG PET/CT (χ2=14.333, P<0.05) , but no significant differences were found in sensitivity and accuracy between the delalyed imaging and standard FDG PET/CT (χ2=0.085 and 0.966, both P>0.05) . The urina?ry bladder activity reached or approached the background during the delayed imaging. Twenty?two malignant lesions of surrounding invasion were confirmed. The positive rate of delayed imaging was higher than that of standard FDG PET/CT:95.5%(21/22) vs 63.6%(14/22);χ2=6.400, P<0.05. Conclusions 18F?FDG PET/CT delayed pelvic imaging after diuresis and hydration is useful for eliminating the artifacts from urina?ry bladder activity, and for detecting the invasion to the rectal surrounding tissue more effectively. It could be a supplement to standard FDG PET/CT.

Objective Tocompare the effects of different waveforms and parameters of electrical stimulation to elicit a blink,and construct a functional electrical stimulation (FES) system to restore synchronous blink in unilateral facial nerve palsy (FNP).Methods Firstly,twenty-four rabbits were surgically induced unilateral FNP and were divided into three groups,who received square,sine and triangle pulse wareforms,respectirely.Both the healthy and the paralysis eyelids of the rabbits received pulse train stimulation to produce a blink in both eyes.For each rabbit,twenty-seven combinations of frequencies (25 Hz,50 Hz and 100 Hz) and nine pulse widths (1-9 ms) were stimulated.The threshold amplitude and electric charge to elicit a blink was compared between different waveforms and different parameters.Secondly,a FES system was constructed to treat six surgically induced unilateral FNP rabbit chosen in the twenty-four rabbits,it consisted by an electromyogram (EMG) amplifier module which record the EMG of the healthy muscle,and a stimulator which received the EMG input and output a pulse train stimulation when triggered by the EMG.Results When the carrier frequency of the pulse train was 25 Hz,it was not able to induce a smooth blink.However,when the carrier frequencies were 50 Hz and 100 Hz,a smooth blink could be induced.The voltage required by 100 Hz was lower than 50 Hz,but it cost more electric charge.The amplitude that square waveforms required was far lower than sine and triangle,but the electric charge between the three waveforms was similar.Synchronous blink could be restored in the six unilateral FNP rabbits with the FES system.Conclusions To elicit a blink,square pulse train delivered in 50 Hz is a preferable option.The motion of the healthy eyelids as a source of information for stimulation of the paralyzed sides can restore the synchronous blink in unilateral FNP rabbits.

Fifty outpatients of newly diagnosed hypertension (NDH,with course of disease ≤ 3 months) and 50 patients with routinely revisit hypertension (RRH,with course of disease ≥ 1 year) were consecutively enrolled respectively from February 2010 to July 2010.They were asked to complete the Beck Depression Inventory (BDI),Spielberger State-Trait Anxiety Inventory (STAI) and the SF-36 questionnaire.The results showed that compared with the RRH group,the NDH group were more worried about being diagnosed as hypertension,dependence and side effects of antihypertensive drugs and complications of hypertension (all P ＜ 0.05).The NDH group displayed significantly higher level of anxiety compared with that of the RRH group (P ＜ 0.001).No significant differences were found in depression and 8 dimensions of SF-36 forms between two groups (P ＞ 0.05).The results indicated that patients of NDH have significant higher level of anxiety than that of RRH,they should be given more psychological intervention and appropriate health education.

Objective To analyze the clinical characteristics of glenoid labral tear combined with rotator cuff tear so as to provide references for arthroscopic surgery.Methods A total of 50 patients (28 males and 22 females)at mean age of 45 years old(ranged from 30 to 58 years)were enrolled in the study.Clinical data including the mechanism and time of injury,symptoms,results of physical examination,X-ray and MRI were collected.Location and degree of rotator cuff and glenoid labral tears found during arthroscopic surgery were also recorded.There were 37 patients with anterior labral tears,eight with superior labrum tears and five with posterior labral tears.The rotator cuff tear was slight in 36 patients,moderate in 10 and severe in three.One patient had extra-large tear.Anterior part of rotator cuff was found torn in 21 patients,middle part in 16 and posterior part in 13.MRI of glenoid labral tears manifested disappearance of triangular appearance and enlarged joint capsule.MRI manifestations of rotator cuff tear were high signal in torn part and continuity loss.Results All patients were followed up for average 36 months(6-72 months),which showed satisfactory results.UCLA score was(15±3)points preoperatively and(32±3)points postoperatively(P＜0.01).Conclusions More attention should be paid to glenoid labral tear to avoid misdiagnosis and delayed treatment,for it is usually combined with rotator cuff tear.Location of rotator cuff tear has some relation to the range of glenoid labral tear,ie,the larger range of glenoid labral tear may induce more posterior part of rotator cuff torn.

Objective To investigate the epidemiological characteristics of three common susceptive gene retared hearing loss in the patients with the congenital deafness in Liuzhou.Methods 161 patients with congenital hearing loss were diagnosed with audiologic evolutions,including newborns and outpatients.The blood samples of all patients were taken for the extraction of DNA which was amplified by PCR.The common mutationsl hot spots of the mitochondrial DNA 12SrRNA,GJB2 and SLC26A4 were examined by restricted enzyme and directed sequencing.Results 1 case(0.62%)was found to carry mitoehondrial DNA 12SrRNA A1555G and 4 patients(2.48%)carried heterozygotes or homozygotes pathologic mutations of GJB2.10 patients(6.21%)were heterozygous carriers with pathologic mutations,IVS7-2 A>G,in the SLC26A4 gene.The detection rate of GJB2,mitochondrial DNA A1555G and SLC26A4 mutations in 161 patients were 9.31%.Conclusion The patients with congenital hearing loss distributed different minority groups in liuzhou zone.The mutational frequencies of the three common gene related hearing loss in the patients of Liuzhou were noticeably lower than the data reported in other regions in China.The gene screening for deafness was very important for early diagnosis and treatment.