Ultrasound-guided core-needle biopsy for peripheral pulmonary lesions: a systematic review and meta-analysis.

AIM: To evaluate the diagnostic value and safety of ultrasound-guided core-needle biopsy for peripheral pulmonary lesions (PPLs). MATERIALS AND METHODS: PubMed, EMBASE, and the Cochrane Library for relevant were searched for studies published up to June 2022. The diagnostic accuracy of US-guided percutaneous transthoracic needle biopsy (PTNB) for the diagnosis of PPLs was evaluated using pooled sensitivity, specificity, diagnostic odds ratio (DOR), positive and negative likelihood ratios (PLR and NLR), and the area under the summary receiver operating characteristic curves value (SROC). RESULTS: The search included 12 original studies (3,830 procedures). For US-guided PTNB, the pooled sensitivity and specificity for the diagnosis of PPLs were 0.93 (95% confidence interval [CI]: 0.91-0.94) and 0.99 (95% CI: 0.96-1.00), respectively. The pooled estimates of the PLR, NLR, and DOR were 134.88 (95% CI: 24.88-731.74), 0.07 (95% CI: 0.06-0.09), and 1,814.95 (95% CI: 333.62-9,873.76), respectively. The area under the SROC curve was 0.95 (95% CI: 0.93-0.97). The overall complication rate was 3.6% (136 of 3,830), including self-limited haemoptysis and asymptomatic pneumothorax, and only six cases of pneumothorax requiring chest tube drainage and one case of severe bleeding were reported. CONCLUSIONS: US-guided core-needle biopsy is an excellent diagnostic tool for PPLs, with high accuracy and excellent technical performance and safety.

Additive Effects of Environmental Enrichment and Ketamine on Neuropathic Pain Relief by Reducing Glutamatergic Activation in Spinal Cord Injury in Rats.

Spinal cord injury (SCI) impairs mobility and often results in complications like intractable neuropathic pain. A multi-approach management of this chronic pain condition has been encouraged, but little has been explored of the field. Here, we focus on the effect and underlying mechanism of environmental enrichment (EE), which promotes voluntary social and physical activities, combined with a clinical analgesic, ketamine, on SCI-induced neuropathic pain as well as motor dysfunction. We performed T13 spinal hemisection in rats, which induced unilateral motor impairment and neuropathic pain-like behaviors in the hindlimb. Treatment regimen started a week after SCI, which consists of ketamine administration (30 mg kg-1 day-1; intramuscular) for 10 days, or EE housing for 20 days, or their combination. Paw withdrawal response to mechanical and thermal stimuli, motor function, burrowing behaviors, and body weight was monitored. Spinal segments at T13 lesion and L4-L6 were collected for histopathological and protein analyses. The joint treatment of EE and ketamine provided greater relief of pain-like behaviors and locomotor recovery than did either paradigm alone. These improvements were associated with reduced cavitation area, astrogliosis, and perilesional phosphorylation of glutamate N-methyl-D-aspartate receptor (NMDAR). Concurrently, lumbar spinal analysis of NMDAR-linked excitatory markers in hypersensitization showed reduced activation of NMDAR, mitogen-activated protein kinase (MAPK) family, nuclear factor (NF)-κB, interleukin (IL)-1ß signaling, and restored excitatory amino acid transporter 2 level. Our data support a better therapeutic efficacy of the combination, EE, and ketamine, in the attenuation of neuropathic pain and motor recovery by reducing spinal glutamatergic activation, signifying a potential multifaceted neurorehabilitation strategy to improve SCI patient outcome.

Alcohol intake and the risk of chronic kidney disease: results from a systematic review and dose-response meta-analysis.

Many prospective cohort studies have investigated the association between the consumption of alcohol and CKD risk and have revealed inconsistent results. In the present study, we aimed to perform a meta-analysis of these studies to assess this association.We searched the PubMed and Embase databases up to 2020 and reviewed the reference lists of relevant articles to identify appropriate studies. We calculated the pooled relative risks with 95% CIs using random effects models, and then performed subgroup and meta-regression analyses. Dose-response meta-analyses were performed by sex separately. We identified 25 eligible prospective cohort studies, including 514,148 participants and 35,585 incident CKD cases. Compared with the category of minimal alcohol intake, light (RR = 0.90, I2 = 49%), moderate (RR = 0.86, I2 = 40%), and heavy (RR = 0.85, I2 = 51%) alcohol intake were associated with a lower risk of CKD. Subgroup meta-analysis by sex indicated that light (RR = 0.92, I2 = 0%), moderate (RR = 0.83, I2 = 39%) and heavy (RR = 0.76, I2 = 40%), alcohol consumption were inversely associated with CKD risk in male. Dose-response meta-analyses detected a nonlinear inverse association between alcohol consumption and the risk of CKD in all participants and linear inverse association in female participants. This meta-analysis shows that light (<12 g/day), moderate (12-24 g/day), and heavy (>24 g/day) alcohol consumption are protective against chronic kidney disease in adult participants especially in males.

Field Ecology and Food Suitability of Tarsonemus spp. (Acari: Tarsonemidae).

Tarsonemus bakeri Ewing (Acari: Tarsonemidae) is a species of mite commonly associated with citrus in many countries including the United States. A short report in 1942 suggested this species is phytophagous, but it has not been reported as a pest in citrus or any other crop since then. A single survey of 78 orchards in three growing regions in California demonstrated that Tarsonemus spp. mites were only associated with leaf samples that had visible sooty mold. A seasonal population study in one citrus orchard showed that all life stages of Tarsonemus spp. were present year-round on leaves and fruit, with the population on fruit reaching a peak in December (59.7 ± 15.2 mites per fruit). Results from a food suitability study showed that the population declined sharply on both plastic and leaf substrate when the mites were not provided a supplementary food source. When supplementary food was provided in the form of Alternaria, honeydew, molasses, or combinations of these, mites survived and multiplied throughout the 29-d study, irrespective of the substrate. Tarsonemus bakeri were found on excised, decaying leaves collected from an orchard. These studies verify that Tarsonemus spp. are associated only with sooty mold in citrus orchards. T. bakeri populations cannot sustain themselves on leaf tissue alone, indicating that they are nondamaging to citrus and therefore need not be considered a phytosanitary concern by importing countries.

Associations between gut microbiota and thyroidal function status in Chinese patients with Graves' disease.

OBJECTIVE: The imbalance of gut microbiota has been linked to manifold endocrine diseases, but the association with Graves' disease (GD) is still unclear. The purpose of this study was to investigate the correlation between human gut microbiota and clinical characteristics and thyroidal functional status of GD. METHODS: 14 healthy volunteers (CG) and 15 patients with primary GD (HG) were recruited as subjects. 16SrDNA high-throughput sequencing was performed on IlluminaMiSeq platform to analyze the characteristics of gut microbiota in patients with GD. Among them, the thyroid function of 13 patients basically recovered after treatment with anti-thyroid drugs (oral administration of Methimazole for 3-5 months). The fecal samples of patients after treatment (TG) were sequenced again, to further explore and investigate the potential relationship between dysbacteriosis and GD. RESULTS: In terms of alpha diversity index, the observed OTUs, Simpson and Shannon indices of gut microbiota in patients with GD were significantly lower than those in healthy volunteers (P < 0.05).The difference of bacteria species was mainly reflected in the genus level, in which the relative abundance of Lactobacillus, Veillonella and Streptococcus increased significantly in GD. After the improvement of thyroid function, a significant reduction at the genus level were Blautia, Corynebacter, Ruminococcus and Streptococcus, while Phascolarctobacterium increased significantly (P < 0.05). According to Spearman correlation analysis, the correlation between the level of thyrotropin receptor antibody (TRAb) and the relative abundance of Lactobacillus and Ruminococcus was positive, while Synergistetes and Phascolarctobacterium showed a negative correlation with TRAb. Besides, there were highly significant negative correlation between Synergistetes and clinical variables of TRAb, TPOAb and TGAb (P < 0.05, R < - 0.6). CONCLUSIONS: This study revealed that functional status and TRAb level in GD were associated with composition and biological function in the gut microbiota, with Synergistetes and Phascolarctobacterium protecting the thyroid probably, while Ruminococcus and Lactobacillus may be novel biomarkers of GD.

[Evaluation and analysis of anxiety, depression and quality of life in vasomotor rhinitis].

Objective: To evaluate the mental state and quality of life in patients with vasomotor rhinitis (VMR) before and after treatment, and to provide guidance for improving the overall health of VMR patients. Methods: Two hundred and twenty VMR patients (VMR group, 118 males, 102 females; aged from 18 to 72 years old), three hundred and twenty allergic rhinitis (AR) patients (AR group, 178 males, 142 females; aged from 18 to 79 years old) from January 2016 to September 2019 were selected in the otolaryngology clinic of Affiliated Hospital of Guizhou Medical University, four hundred and twenty-three healthy people (control group, 243 males, 180 females; aged from 19 to 70 years old) were selected in physical examination center at the same time by continuous enrollment method, symptom check list (SCL-90), self-rating anxiety scale (SAS) and self-rating depression scale (SDS) were used to evaluate the mental state of VMR patients before and after treatment, and 12-item short form health survey version 2.0 (SF-12v2) was used to evaluate their quality of life, statistical data were collected and analyzed by ANOVA and t-test. Results: The scores of eight factors (physical function, role physical function, general health, vitality, role-emotional, mental health) of SF-12v2 in VMR patients before treatment were lower than that of posttreatment, that of AR patients and the control group, the differences were significant (all P<0.05), the scores of somatization, obsession, depression, anxiety and psychosis in SCL-90 in VMR patients before treatment were significantly higher than that of posttreatment, that of AR patients and the control group (all P<0.05), the SAS and SDS in VMR patients before treatment (51.28±16.32; 53.28±18.55) were significantly higher than that of posttreatment (38.53±13.21; 39.35±13.34), that of AR patients (42.23±14.32; 43.32±13.78) and the control group (29.78±10.07;33.46±10.55; t(SAS) were 9.007, 6.813 and 20.59; t(SDS) were 9.043, 7.154 and 17.260, all P<0.05). Conclusion: VMR patients generally suffer from psychological damage, which seriously affects the quality of life of the patients. On the basis of routine treatment, we should attach more importance to the negative psychology of VMR patients and intervene when necessary.

Direct Detection Constraints on Dark Photons with the CDEX-10 Experiment at the China Jinping Underground Laboratory.

We report constraints on the dark photon effective kinetic mixing parameter (κ) with data taken from two p-type point-contact germanium detectors of the CDEX-10 experiment at the China Jinping Underground Laboratory. The 90% confidence level upper limits on κ of solar dark photon from 205.4 kg-day exposure are derived, probing new parameter space with masses (m_{V}) from 10 to 300 eV/c^{2} in direct detection experiments. Considering dark photon as the cosmological dark matter, limits at 90% confidence level with m_{V} from 0.1 to 4.0 keV/c^{2} are set from 449.6 kg-day data, with a minimum of κ=1.3×10^{-15} at m_{V}=200 eV/c^{2}.

[Correlation between serum bilirubin and cardiovascular autonomic neuropathy in patients with type 2 diabetes].

Objective: To investigate the correlation between serum bilirubin and cardiovascular autonomic neuropathy (CAN) in type 2 diabetes mellitus patients. Methods: A total of 369 patients with type 2 diabetes mellitus who were hospitalized at the Department of Endocrinology, Nanjing Jinling Hospital from April 2017 to October 2018 were enrolled, including 226 males and 143 females, with an average age of (54.6±12.1) years. According to cardiovascular reflex tests (CARTs), all the patients were divided into Non CAN group(149 patients without CAN) and CAN group (220 patients complicated with CAN). The difference of serum bilirubin levels between the two groups was compared. The differences of CARTs and the incidence of CAN were compared by tertiles of serum bilirubin levels. The binary logistic regression was used to analyze the risk factors for diabetic cardiovascular autonomic neuropathy. Results: The serum total bilirubin [(9.28±2.74) µmol/L vs (11.08±2.98) µmol/L, P<0.001], direct bilirubin [(3.17±1.20) µmol/L vs (3.71±1.24) µmol/L, P<0.001] and indirect bilirubin levels [(6.11±1.89) µmol/L vs (7.37±2.10) µmol/L, P<0.001] in CAN group were significantly lower than that in Non CAN group. With the increase of serum bilirubin, the incidence of CAN decreased (P<0.01). Multivariate Logistic regression analysis showed that serum total bilirubin (OR=0.819, 95%CI: 0.744-0.901, P<0.001), direct bilirubin (OR=0.739, 95%CI: 0.601-0.908, P=0.004) and indirect bilirubin (OR=0.749, 95%CI: 0.653-0.860, P<0.001) were inversely correlated with the incidence of CAN. Conclusions: Within the physiological range, lower level of serum bilirubin is inversely correlated with the incidence of CAN. It is noteworthy to screen diabetic cardiovascular autonomic neuropathy in patients with type 2 diabetes mellitus who had a lower serum bilirubin level.

Diminutive Incision Acromioplasty Assisted with Arthroscopy in the Treatment of Chinese Patients with Subacromial Impingement Syndrome / Acromioplastia de Incisión Diminuta Asistida con Artroscopia en el Tratamiento de Pacientes Chinos con Síndrome de Pinzamiento Subacromial

ABSTRACT Background: Many causes can lead to shoulder pain and subacromial impingement syndrome (SIS) is the most frequently recorded disorders. The aim of this study was to evaluate the clinical effects of diminutive incision acromioplasty assisted with arthroscopy for the treatment of Chinese patients with subacromial impingement syndrome. Subject and Methods: Twenty-two patients with 24-painful shoulders subacromial impingement syndrome were enrolled. All painful shoulders were in Grades II (8) and III (16) according to Neer's classification. Detailed physical examination was performed. Conventional radiography and subsequent magnetic resonance imaging (MRI) of the shoulder region of all patients were done. The University of California at Los Angeles Shoulder (UCLA) score system was used for all patients to evaluate their satisfaction after surgery. The preoperative recordings of the UCLA scores were collected and all enrolled cases including 24-painful shoulders were available for follow-up in 1, 3, 6, 12 months after surgery. Results: According to the UCLA scoring system, the symptom of all painful shoulders were improved after one year postoperatively. The average score before surgery from 15.4 points increased to 31.2 points postoperatively, showing a statistical difference (p < 0.05). Conclusions: A diminutive incision acromioplasty assisted with arthroscopy is a reliable approach to treat Chinese patients with subacromial impingement syndrome. All painful shoulders were obviously improved in one year after surgery.

ABSTRACT Antecedentes: Muchas causas pueden provocar dolor de hombro y síndrome de compresión subacromial (SIS) es el trastorno más frecuentemente registrado. El objetivo de este estudio fue evaluar la clínica. Efectos de la acromioplastia con incisión diminuta asistida con artroscopia para el tratamiento de Pacientes chinos con síndrome de pinzamiento subacromial. Sujeto y métodos: Se incluyeron veintidós pacientes con síndrome de afectación subacromial de 24-hombros dolorosos. Todos los hombros dolorosos estaban en Grados II (8) y III (16) de acuerdo con la clasificación de Neer. Se realizó examen físico detallado. Se realizaron radiografías convencionales y, posteriormente, imágenes de resonancia magnética (IRM) de la región del hombro de todos los pacientes. El sistema de puntuación de la Universidad de California en Los Angeles Shoulder (UCLA) se utilizó para que todos los pacientes evaluaran su satisfacción después de la cirugía. Los registros preoperatorios de las puntuaciones de UCLA se recopilaron y todos los casos incluidos, incluidos 24-hombros dolorosos, estaban disponibles para el seguimiento en 1, 3, 6 y 12 meses después de la cirugía. Resultados: De acuerdo con el sistema de puntuación de UCLA, el síntoma de todos los hombros dolorosos mejoró después de un año después de la operación. La puntuación promedio antes de la cirugía de 15.4 puntos aumentó a 31.2 puntos después de la operación, mostrando una diferencia estadística (p < 0.05) Conclusiones: Una acromioplastia de incisión diminuta asistida con artroscopia es un enfoque confiable para tratar a pacientes chinos con síndrome de pinzamiento subacromial. Todas las lesiones dolorosas se mejoraron obviamente en un año después de la cirugía.

Adiponectin regulates thermal nociception in a mouse model of neuropathic pain.

BACKGROUND: Adiponectin, a cytokine secreted by adipocytes, plays an important role in regulating glucose and lipid metabolism. However, the role of adiponectin in pain conditions is largely unknown. This study aimed to identify the role and mechanism of adiponectin in nociceptive sensitivity under physiological and pathological states utilising adiponectin knockout (KO) mice. METHODS: Wild type (WT) and adiponectin KO mice were subjected to partial sciatic nerve ligation (pSNL) or sham operation. Pain-like behavioural tests, including thermal allodynia, hyperalgesia, and mechanical allodynia, were performed before and after pSNL from Day 3-21. Dorsal root ganglions (DRGs), lumbar spinal segments at L3-5, and somatosensory cortex were collected for protein measurement via western blotting and immunofluorescence staining. RESULTS: Compared with WT mice, KO mice had significantly lower (40-50%) paw withdrawal latency to innocuous and noxious stimuli before and after pSNL. In DRG neurones from KO mice, where adiponectin receptor (AdipoR) 2 is located, phosphorylated p38 mitogen-activated protein kinase (p-p38 MAPK) and heat-sensitive transient receptor potential cation channel subfamily V member 1 (TRPV1) were significantly higher (by two- to three-fold) than from WT mice. In spinal microglia and somatosensory cortical neurones, where AdipoR1 is mainly located, p-p38 MAPK and TRPV1 were also higher (by two- to three-fold) in KO compared with WT mice, and altered signalling of these molecules was exacerbated (1.2- to 1.3-fold) by pSNL. CONCLUSIONS: Our results show that adiponectin regulates thermal nociceptive sensitivity by inhibiting activation of DRG neurones, spinal microglia, and somatosensory cortical neurones in physiological and neuropathic pain states. This study has relevance for patients with adiponectin disorders, such as obesity and diabetes.

[Three-dimensional architecture of intraosseous vascular anatomy of the hamate: a micro-computed tomography study].

OBJECTIVE: To obtain three-dimensional intraosseous artery of the hamate and to provide the vascular anatomy basis of hamate fracture fixation. METHODS: PbO (lead monoxide, Sinopharm Chemical Reagent Beijing Co. Ltd) was ground into particles less than 40 µm and suspended in turpentine oil (Chemical Reagent Beijing Co. Ltd) at ratios of 1 g : 1.5 mL, 1 g : 1 mL and 1 g : 0.5 mL. Three specimens were investigated. Brachial arteries were cannulated and perfused with lead-based contrast agent. Hamates were harvested and scanned using micro-computed tomography (microCT). The acquisition protocols were as follows: CT scan setup: total rotation [Degrees], 360; rotation steps, 360; X-ray detector setup: transaxial, 2048; axial, 2048; exposure time, 1 500 ms, Binning, 1; system magnification: high-med. X-ray tube setup: 80 kV, 500 mA current. The down-sampling factor used in the reconstruction was 2. The effective voxel size of the final image was 27.30 µm. The three-dimensional model of the hamate was generated and the distribution and pattern of vessels were evaluated. RESULTS: There were abundant extraosseous vessels around the hamate. They were mainly running in the tendons and ligaments around the hamate. Four vascular zones were identified on the hamate surface. They were on the palmar platform of the hamate body, on the dorsal side, on the ulnar side and on the tip of hamulus, namely. There were anastomoses among 4 vascular zones. We did not observe any vessels penetrating through the articular cartilage. The extraosseous vessels of the vascular zones gave a number of intraosseous branches into the hamate. The hamate body received intraosseous blood supply from the dorsal, palmar and ulnar while the hamulus from the palmar, ulnar and hamulus tip. There were some intraosseous branches anastomosing with each other. CONCLUSION: The extraosseous and intraosseous vessels of the hamate were more than what used to be considered. The hamate body and hamulus received blood supply from multiple directions and arteries anastomosed extensively both outside and inside the hamate, making it possible that the intraosseous perfusion survived after fracture. It is likely that the nonunion after the hamate fracture is not caused by the vascular damage but the malalignment of the fragments.

Food Suitability and Population Dynamics of Lorryia formosa (Acari: Tydeidae).

Lorryia formosa Cooreman (Acari: Tydeidae) is a species of mite commonly associated with citrus in many countries including the United States. A survey report in 1957 suggested phytophagous nature, while other studies claimed that L. formosa populations are associated with honeydew producing insects and sooty mold and it acts as a sanitizing agent. We investigated the effect of various diets on the survival and progeny production of L. formosa on excised leaves and the survival and potential to cause feeding damage to leaves of potted plants in a greenhouse study. A 2-yr field survey of a mandarin orchard was also conducted to elucidate the seasonal infestation, damage potential and population structure of L. formosa in a natural habitat. Results showed that all L. formosa adults and immatures died in less than 14 d on excised leaves, did not survive beyond 7 d on potted citrus plants alone, and caused no observable feeding damage to leaves or fruit. When sugar water, honeydew, or cottony cushion scale, Icerya purchasi Maskell (Hemiptera: Margarodidae), was present, adults and immatures survived the duration of the experiments and produced additional generations. The field survey showed that all stages of L. formosa were present in a mandarin orchard throughout the year and insecticide applications affected but did not eliminate mite populations. Fruit generally had a greater percentage infestation of mites (44.8 ± 4.0) than leaves (16.0 ± 4.7). These studies confirmed that L. formosa cannot sustain a population on leaf tissue alone and is nondamaging to citrus in California.

[Study on the expression of epidermal growth factor receptor protein during benzo(a)pyrene induced carcinogenesis].

Objective: To explore the expression of epidermal growth factor receptor(EGFR) protein during benzo(a)pyrene (BaP) induced carcinogenesis. Methods: This study, we firstly utilized immunofluorescence assay and Western-blot to examine EGFR expression of the BaP which was constructed previously by project team induced malignant transformation human bronchial epithelial cell (BTC) and the control (16HBE cell). Then, we selected 36 healthy SD rats, divided into two groups according to simple random method, 18 rats each group. The constructed rat lung neoplasm model induced by pulmonary injection of BaP (10 mg/ml of BaP solution in 0.2 ml corn oil), contrast group use 0.2 ml corn oil, lung tissue was collected and the EGFR expression of lung tissue was detected by immunofluorescence assay and Western blot. T analysis was used to test the different of EGFR between two groups. Results: Immunofluorescence analysis showed that the EGFR expression in BTC was significantly higher than 16HBE cell. Meanwhile, Western blot also was used to confirmed this result, the relative expression of EGFR protein in the rats of the model group the control group were 1.04±0.13 and 2.32±0.12, respectively, and the difference was statistically significance (t=12.39, P<0.001). In vivo, well-defined tumor was found in the rat with pulmonary injection of BaP, and the lung showed diffuse alveolar septal thickening, alveolar wall destruction and pulmonary alveoli fusion, which suggested that the rat lung neoplasm model was constructive successfully. Furthermore, we found the EGFR expression of lung was increased dramatically in the rat lung neoplasm model by immunofluorescence detection and Western blot. The relative expression of EGFR protein in the rats of the model group the control group were 0.21±0.03 and 1.30±0.07, respectively, and the difference was statistically significance (t=12.84, P<0.001). Conclusion: Expression of EGFR protein was increased during BaP carcinogenesis, and EGFR may play an important role in the carcinogenesis of BaP.

Pot1 OB-fold mutations unleash telomere instability to initiate tumorigenesis.

Chromosomal aberrations are a hallmark of human cancers, with complex cytogenetic rearrangements leading to genetic changes permissive for cancer initiation and progression. Protection of Telomere 1 (POT1) is an essential component of the shelterin complex and functions to maintain chromosome stability by repressing the activation of aberrant DNA damage and repair responses at telomeres. Sporadic and familial mutations in the oligosaccharide-oligonucleotide (OB) folds of POT1 have been identified in many human cancers, but the mechanism underlying how hPOT1 mutations initiate tumorigenesis has remained unclear. Here we show that the human POT1's OB-folds are essential for the protection of newly replicated telomeres. Oncogenic mutations in hPOT1 OB-fold fail to bind to single-stranded telomeric DNA, eliciting a DNA damage response at telomeres that promote inappropriate chromosome fusions via the mutagenic alternative non-homologous end joining (A-NHEJ) pathway. hPOT1 mutations also result in telomere elongation and the formation of transplantable hematopoietic malignancies. Strikingly, conditional deletion of both mPot1a and p53 in mouse mammary epithelium resulted in development of highly invasive breast carcinomas and the formation of whole chromosomes containing massive arrays of telomeric fusions indicative of multiple breakage-fusion-bridge cycles. Our results reveal that hPOT1 OB-folds are required to protect and prevent newly replicated telomeres from engaging in A-NHEJ mediated fusions that would otherwise promote genome instability to fuel tumorigenesis.

Repair of critical-sized bone defects with anti-miR-31-expressing bone marrow stromal stem cells and poly(glycerol sebacate) scaffolds.

The repair of critical-sized defects (CSDs) is a significant challenge in bone tissue engineering. Combining the use of progenitor cells with gene therapy represents a promising approach for bone regeneration. MicroRNAs play important roles in most gene regulatory networks, regulate the endogenous expression of multiple growth factors and simultaneously modulate stem cell differentiation. Our previous study showed that knocking down miR-31 promotes the osteogenesis of bone marrow stromal stem cells (BMSCs). To investigate the therapeutic potential of cells engineered to express anti-miR-31 for CSD repair, lentiviral vectors encoding negative control, miR-31 precursor and anti-sense sequences were constructed and transduced into osteo-inductive BMSCs. The expression of osteogenic-specific genes, alkaline phosphatase activity and Alizarin Red S staining were investigated to evaluate the effects of miR-31 on the cell fate of BMSCs over a 3-week period. In addition, miR-31-modified BMSCs seeded on poly(glycerol sebacate) (PGS) scaffolds were used to repair 8 mm critical-sized calvarial defects in rats. The results showed that miR-31 suppression significantly increased the expression of osteogenic-specific genes in vitro at the mRNA and protein levels, and that robust new bone formation with high local bone mineral density was observed in the anti-miR groups in vivo. Moreover, the PGS scaffolds carrying anti-miR-31-expressing BMSCs exhibited good biocompatibility and a high regeneration rate (~60%) within in vivo bone defects. Our results suggest that miR-31 gene delivery affects the potential of BMSCs for osteogenic differentiation and bone regeneration and that PGS is a potential substrate for genetically modified, tissue-engineered bone in the repair of large bone defects.

Induction of retinal ganglion-like cells from fibroblasts by adenoviral gene delivery.

Central nervous system neurons fail to regenerate after birth, which greatly hampers the effective treatment of many neurodegenerative diseases. Neurons differentiated from induced pluripotent stem cells have been considered a possible option for cell-based therapies. Recent discoveries have revealed that fibroblasts can be directly converted into neurons without a transition through a pluripotent state. This approach might serve as a more efficient and convenient method for the cellular therapy of neurodegenerative diseases. Currently, several types of neurons have been directly generated from fibroblasts, including dopamine neurons, motor neurons and neural progenitor cells. In our study, by screening a series of candidate genes, we found that the adenovirus-mediated transduction of Ascl1, Brn3b and Ngn2 can directly convert mouse fibroblasts to retinal ganglion-like cells. The induced retinal ganglion-like cells co-express multiple retinal ganglion cell markers, and exhibit membrane properties of functional neurons. The reprogramming mediated by adenoviruses occurs much sooner than that mediated by lentiviruses. Furthermore, the induced retinal ganglion-like cells that are produced via adenoviral gene delivery are free of exogenous gene integration. Retinal ganglion-like cells that are induced by adenoviruses demonstrate great potential applicability in clinical therapy and provide a novel platform for the research of retinal degenerative diseases.

SP600125, a JNK inhibitor, suppresses growth of JNK-inactive glioblastoma cells through cell-cycle G2/M phase arrest.

SP600125 is a well studied inhibitor of c-Jun N-terminal kinase (JNK). Its direct biochemical effects on JNK-inactive tumor cells are usually ignored. In this study, we investigated the effects of SP600125 on JNK-inactive U251 human glioblastoma cells. Our results demonstrate that, 20 microM or more SP600125 can induce significant cell growth inhibition and cell-cycle G2/M phase arrest in U251 cells. Interestingly, we also found that SP600125 can stop the duplicated chromosomes from separating into two cells and the karyokinesis progression. Our study opened up a new perspective for further studies involved in JNK inhibitors or anti-glioma therapy.

Improvement of glucose tolerance by rhein with restored early-phase insulin secretion in db/db mice.

AIMS: In the present study, we investigated whether rhein exerted hypoglycemic action and rhein's effect on the pancreatic ß cell in db/db mice. MATERIALS AND METHODS: Thirty 4-week-old db/db mice were randomized to treatment with rhein (120 mg/kg) (no.=15) and placebo (1% natrium cellulose solution) (no.=15) for 8 weeks, respectively. Fifteen age-matched non-diabetic littermates db/m mice treated with placebo were studied as non-diabetic control. After an 8-week treatment, ip glucose tolerance test (IPGTT) and arginine tolerance test were performed. Area under curve (AUC) of insulin levels in IPGTT was calculated to evaluate insulin secretory function. Immunohistochemical staining of insulin was performed to estimate ß cell mass. TUNEL assay was performed to determine ß cell apoptosis. Islet isolation and perifusion were performed to evaluate kinetics of insulin release in vitro, especially first-phase insulin. RESULTS: Compared with control group, AUC of glucose concentrations significantly decreased in the rhein-treated group (p<0.05). Simultaneously, AUC of insulin levels increased in the rhein-treated group (p<0.05), especially in the first 30 min after glucose load. Perifusion showed that the rhein-treated group manifested a significantly increase of first-phase insulin secretion. Immunohistochemical study and TUNEL assay showed that rhein treatment greatly preserved ß cell mass and inhibited ß cell apoptosis. CONCLUSIONS: Rhein treatment significantly improved glucose- dependent and independent insulin secretion by preservation of ß cell mass and inhibition of ß cell apoptosis in db/db mice. The characteristics of rhein may make it a novel therapeutic means for preventing from or curing diabetes in the near future.

Association of leptin receptor gene polymorphisms and essential hypertension in a Chinese population.

BACKGROUND: The leptin receptor (LEPR) is an important regulator of leptin activity and resistance. Several single nucleotide polymorphisms (SNP) of LEPR have been linked to diseases accompanying obesity and/or obesity-related diseases in different populations. However, the results from published studies remain inconsistent rather than conclusive. AIM: To investigate whether LEPR SNP are associated with essential hypertension and related metabolic traits in Chinese subjects. MATERIALS AND METHODS: A total of 544 Chinese patients with hypertension and 357 non-hypertensive subjects were screened. The genotypes of LEPR polymorphisms were determined by PCR-restriction fragment length polymorphism methods. Demographic and biochemical characteristics including waist circumference, waist-to-hip ratio, body mass index (BMI), lipids profiles, glucose metabolism, and leptin levels were obtained for analysis. RESULTS: This case-control study showed associations between the frequencies of AA genotype and A allele of Gln223Arg and hypertension (p=0.029, p=0.002, respectively). Furthermore, the Gln223Arg polymorphism was significantly associated with plasma leptin levels (p<0.001), while no correlations between Lys109Arg SNP and hypertension were found. Multivariate logistic regression analysis evidenced that A allele carriers of Gln223Arg (AA+AG) showed higher risks of hypertension than GG carriers after adjustment of age and sex (adjusted odds ratio: 1.549, 95% confidence interval: 1.031- 2.036, p=0.035). BMI, fasting serum insulin, oral glucose tolernace test (OGTT)-2h glucose, serum leptin, as well as LDL-cholesterol (LDL-C) levels were also independent risk factors of hypertension in this population. In addition, significant associations were observed between the Gln223Arg and Lys109Arg SNP and serum total cholesterol, LDL-C, and fasting plasma glucose levels in hypertensive patients. Besides, A allele of Gln223Arg had raised diastolic blood pressure, compared with GG carriers (p=0.001). While variance of Lys109Arg was associated with waist-to-hip ratio, OGTT-2h glucose, and homeostasis model assessment of insulin resistance (p<0.05). CONCLUSIONS: LEPR polymorphisms may be a marker for susceptibility to essential hypertension in Chinese subjects, and be involved in the development of several features including dyslipidemia and impaired glucose regulation in hypertension subjects.

Improved glucose-stimulated insulin secretion by intra-islet inhibition of protein-tyrosine phosphatase 1B expression in rats fed a high-fat diet.

BACKGROUND: Insulin resistance of pancreatic ß-cell itself may be a potential link between systemic insulin resistance and impaired insulin secretion in Type 2 diabetes. Protein tyrosine phosphatase 1B (PTP1B) dephosphorylates tyrosine residues in insulin receptors (IR) and IR substrate (IRS) proteins, and thereby inhibits insulin signaling. Thus the impact of PTP1B expression on ß-cell insulin pathway may affect insulin secretory function. AIM: The aim of the present study was to investigate the effects of intra-islet inhibition of PTP1B expression on glucose-stimulated insulin secretion and potential mechanisms in rats fed a high-fat diet (HFD). MATERIALS AND METHODS: Twenty 10-week-old Sprague Dawley rats were randomly assigned to a regular diet (RD) or a HFD for 8 weeks. At the end of the 8th week, fasting glucose, fasting insulin concentration and lipid profile were measured and an oral glucose tolerance test was done after 12-h fast. Then islet isolation was performed for static incubation and perifusion. Recombinant adenoviruses containing siPTP1B (Ad-siPTP1B), or siControl (Ad-siControl) sequences were constructed using AdEasy™ system. Islets were transfected and then assigned to the Ad-siPTP1B group, the Ad-siControl group, and mock control group. Real-time RT-PCR and Western blot were used to evaluate the expression level of PTP1B. Western blot of glucose transporter 2 (GLUT-2) and glucokinsase were also done to investigate the ß-cell glucose-sensing apparatus. Islets were incubated with Krebs-Ringer bicarbonate containing 2.8 mmol/l glucose then 16.7 mmol/l glucose to evaluate glucose-stimulated insulin secretion (GSIS). Islet perifusion was also performed to evaluate kinetics of insulin release in vitro. RESULTS: HFD rats manifested modest glucose intolerance compared with RD group. And PTP1B expression in isolated islets of rats in the HFD group was higher than that of the RD group. GSIS was impaired in islets of HFD rats (2.3±0.5-fold as basal for HFD vs 8.1±1.3-fold for RD; p<0.05). Ad-siPTP1B treatment resulted in 73% decrease in PTP1B mRNA levels and 61% decrease in PTP1B protein compared with islets treated with Ad-siControl (p<0.05). Simultaneously, PTP1B inhibition resulted in 4.7±0.8-fold increase of GSIS from basal (vs 1.9±0.1-fold for Ad-siControl, p<0.05). Perifusion showed notable improvement of first-phase insulin secretion by AdsiPTP1B treatment. Significant decrease of both GLUT-2 (by 49.8%) and glucokinase (GCK, by 43.7%) were found in the HFD group when compared with the RD group, while up-regulation of both GLUT-2 (by 98%) and GCK (by 62%) was achieved after PTP1B inhibiton by Ad-siPTP1B. CONCLUSIONS: Intra-islet PTP1B is an important physiological regulator of glucose-induced insulin release and the characteristics of PTP1B inhibitors in insulin secretion could make it a potential novel therapeutics for protection of ß-cell secretory function in Type 2 diabetes.