[Clinicopathological characteristics of SMARCB1(INI1)-deficient sinonasal carcinoma].

Objective: To investigate the clinicopathological characteristics, diagnosis, differential diagnosis and prognostic factors of SMARCB1 (INI1)-deficient sinonasal carcinoma (SDSC). Methods: Sixteen cases of SDSC diagnosed in the Department of Pathology, Beijing Tongren Hospital from January 2016 to September 2020 were enrolled. Ninety-nine cases of small round cell malignant tumors of the head and neck were selected as the control, including poorly-differentiated squamous cell carcinoma (n=10), poorly-differentiated adenocarcinoma (n=5), undifferentiated carcinoma (SNUC, n=4), NUT carcinoma (n=5), neuroendocrine carcinoma (n=10), and other non-epithelial tumors [olfactory neuroblastoma (n=10), rhabdomyosarcoma (n=10), NK/T-cell lymphoma (n=10), malignant melanoma (n=10), Ewing's sarcoma/primitive neuroectodermal tumor (EWS/PNET, n=5)] and non-keratinizing undifferentiated nasopharyngeal carcinoma (n=20). The clinical and pathologic characteristics of SDSC, and immunohistochemical (IHC) expression of broad-spectrum CKpan, CK7, CK8/18, CK5/6, p63, p40, p16, INI1, NUT and neuroendocrine markers (Syn, CgA, CD56) were evaluated. In situ hybridization (ISH) was used to detect EBER and fluorescence in situ hybridization (FISH) to detect INI1 gene deletion. Results: The 16 cases of SDSC accounted for 1.3% (16/1 218) of all malignant sinonasal tumors in the author's unit during this time period, and 2.4% (16/657) of all malignant epithelial tumors. Microscopically, there was no clear squamous and adenomatous differentiation, but "rhabdoid-like" cells, are often seen. All SDSC cases were positive for CKpan and CK8/18, negative for INI1; Epstein-Barr virus was not detected by ISH; and INI1 gene deletion was observed in all 11 SDSC patients with FISH. Twelve cases were followed up for 3-47 months. One died of tumor-related diseases half a year after diagnosis, and the remaining patients were alive with tumor, the longest survival time was 47 months. Conclusion: SDSC should be differentiated from a variety of poorly-differentiated tumors in the sinonasal area. Histologically, SDSC has no clear differentiation, but the tumor cells are characteristically basal-like or rhabdoid-like, with non-specific vacuoles, translucent or vacuolar nuclei, prominent nucleoli and necrotic foci. They are negative for INI1 IHC staining, and FISH demonstrates INI1 gene deletion. The clinical prognosis is still unclear, further studies on its biologic behavior and treatment methods are warranted.

[Ruptured chordae tendineae of tricuspid valve in neonate with intractable persistent pulmonary hypertension: a case report and literature review].

Objective: To analyze the clinical characteristics and treatment of tricuspid valve prolapse caused by chordal rupture complicated with persistent pulmonary hypertension in neonates. Methods: The clinical data of a male neonate with tricuspid valve prolapse complicated with persistent pulmonary hypertension admitted to the Neonatal Intensive Care Unit of Children's Hospital of Hebei Province in November 2018 was analyzed retrospectively. Related literature up to September 2020 was searched with the strategy of "(neonate OR newborn) AND (tricuspid valve prolapse) AND (rupture OR necrosis) AND (papillary muscle OR chordae tendineae) AND (pulmonary hypertension)" in Wanfang, CNKI and PubMed database in Chinese and English. The characteristics of the disease were summarized. Results: A male full-term neonate was admitted due to presenting severe cyanosis for 9 hours. He was born by caesarean section and presented severe cyanosis and dyspnea at 10 min of ages, unresponsive to the positive airway pressure resuscitation. After 9 hours of mechanical ventilation, there was no improvement. Thus he was transferred to Children's Hospital of Hebei Province. On admission, the initial blood gas analysis showed an arterial partial pressure of oxygen of 22.5 mmHg (1 mmHg=0.133 kPa). The echocardiography revealed prolapsed anterior leaflet of tricuspid valve, severe tricuspid regurgitation (TR) and pulmonary artery hypertension, and right to left shunt via a patent foramen ovale. The arterial duct was closed. The chest X-ray was normal. The boy was treated with nitric oxide, milrinone, and continued mechanical ventilation initially. Addition of prostacyclin analog (treprostinil) on day 3 led to significant improvement of pulmonary blood flow, oxygenation, and stabilization, so that the extracorporeal membrane oxygenation therapy was avoided. At 11 months after birth, the boy underwent cardiac surgery. At surgery, the rupture of chordal tendineae in anterior leaflet of tricuspid valve was found. Tricuspid annuloplasty, valvuloplasty and repair of patent foramen ovale were successfully performed. The follow-up echocardiogram at postoperative 3 months showed only mild tricuspid insufficiency. The boy was well at last follow-up at 22 months of age with normal cognitive skill development. According to literature, 20 cases of papillary muscle or chordae tendineae rupture in neonates had been reported in 12 English papers. Among the total 21 neonates, there were 12 male infants and only one premature infant with gestational age of 33 weeks. They presented with profound cyanosis soon after birth. All of them received endotracheal intubation and mechanical ventilation. Other treatments included inhalation of nitric oxide, intravenous milrinone, vasoactive drugs, diuretics and prostacyclin, etc. Extracorporeal membrane oxygenation (ECMO) was used in 6 infants as a bridge to surgical treatment. Two cases reported earlier death of cardiopulmonary failure without operation and the rest 19 survived after surgery. The followed surgery or autopsy revealed that all of them had tricuspid valve prolapse, rupture of papillary muscle or chordae tendineae. Conclusions: The severe TR resulting from rupture of papillary muscle or chordate tendineae in neonates is rare and could cause severe hypoxemia. Early recognition, adequate cardiopulmonary support to stabilize the hemodynamic status and timely surgery can significantly reduce the mortality.

[Diagnosis of adult Philadelphia chromosome-like acute lymphoblastic leukemia by fluorescence in situ hybridization].

Objective: To establish a screening system of adult Philadelphia chromosome-like acute lymphoblastic leukemia (Ph-like ALL) by fluorescence in situ hybridization (FISH) . Method: Based on the genetic characteristics of Ph-like ALL, FISH probes were designed for ABL1, ABL2, JAK2, EPOR, CRLF2, CSF1R, PDGFRB, and P2RY8 gene breakpoints, which were used to screen Ph-like ALL in B-ALL patients without BCR-ABL1, ETV6-RUNX1, MLL, and E2A gene arrangement. Furthermore, it was analyzed in combination with flow immunophenotype, next-generation sequencing for targeted gene mutations, and RNA sequencing (RNA-seq) . Results: A total of 189 adult B-ALL patients diagnosed in Nanfang Hospital from January 2016 to April 2019 were enrolled in this study. Using FISH and/or PCR, BCR-ABL1, ETV6-RUNX1, MLL, or E2A arrangement was detected in 83 of them, and Ph-like ALL was detected by FISH in the other 106, resulting in the presence of typical gene arrangements of Ph-like ALL in 12 patients (11.3% , 12/106) . Validated by RNA-seq, the sensitivity and specificity of FISH for Ph-like ALL were 71.4% and 95.8% , respectively. After further analysis with immunophenotype, targeted gene mutations, and RNA-seq, 14 (13.2% , 14/106) were diagnosed with Ph-like ALL. Conclusion: This data shows high specificity of FISH for identification of Ph-like ALL and combining immunophenotype and sequencing technology can improve the diagnostic system.

Prediction of prostate cancer aggressiveness with a combination of radiomics and machine learning-based analysis of dynamic contrast-enhanced MRI.

AIM: To investigate whether the combination of radiomics and automatic machine learning-based classification of original images from multiphase dynamic contrast-enhanced (DCE)-magnetic resonance imaging (MRI) can predict prostate cancer (PCa) aggressiveness before biopsy. MATERIALS AND METHODS: Forty consecutive biopsy-confirmed PCa patients were included. Biopsy was performed within 4 weeks after the DCE-MRI examinations. According to the time-signal-intensity curve, lesion segmentation was performed on the first and on the strongest phase of the enhancement on the original DCE-MRI images, and 1,029 quantitative radiomics features were calculated automatically from each lesion, wherein there were three datasets available (Dataset-F, Dataset-S and Dataset-FS). The variance threshold method, select k-best method and least absolute shrinkage and selection operator (LASSO) algorithm were used to reduce the feature dimensions. Five machine learning approaches leveraging cross-validation were employed, and the clinical value of each model was evaluated by area under the receiver operating characteristic curve (AUC). Correlation analysis was performed between the features of the machine learning model that achieved the best classification performance and the Gleason score (GS) of the PCa lesion. RESULTS: Eight, four, and 16 features were selected as optimal subsets in Dataset-F, -S and -FS, respectively. Among all three datasets, logistic regression (LR)-based analysis with Dataset-FS had the highest predication efficacy (AUC=0.93). Ten features in Dataset-FS showed significantly positively correlation with GS. The model performance of Dataset-F was generally better than that in Dataset-S. CONCLUSIONS: A combination of radiomics and machine learning-analysis based analysis of the union of the first and strongest phases of original DCE-MRI images can predict PCa aggressiveness non-invasively, accurately, and automatically.

[Combined use of the Solitaire AB stent and Navien delivery catheter for mechanical thrombectory of acute vertebrobasilar artery occlusion].

Objective: To investigate the efficacy and safety of mechanical thrombectory with a tri-axial system of the solitaire AB stent through a Navien delivery catheter to treatment acute vertebrobasilar artery occlusion. Methods: The clinical data of 16 consecutive patients with acute ischemic stroke of vertebrobasilar artery occlusion treated with the Solitaire AB device from March 2016 to March 2018 in Department of Neurosurgery, the First Affiliated Hospital, Medical College of Shihezi University, were extracted and then retrospectively analyzed.Recanalization rate as well as complications after treatment were analysized. Also, neurological functions of the patients before and after operation 1 weeks, measured by National Institute of Health stroke scale (NIHSS) score were compared via t test and the clinical outcomes were assessed by modified Rankin score (mRS) at 90 days after operation. Results: There were 10 male and 6 female patients with a median age of 59 years (ranging from 45 to 78 years).Fourteen patients resulted in successfully recanalization, and 2 cases failed both of whose onset to sheath mean time were 8.5 hours.The NIHSS score at 7 days was 13±7, which was significantly decreased compared to the admission NIHSS score 24±4 (P=0.000).No symptomatic intracranial hemorrhage case was found after operation.At 90 days, 8 patients achieved good outcome (mRS 0 to 2), one patient died (mRS 6),two patients moderately seriously disabled (mRS 4).Three patients resulted in moderate outcome (mRS 3).In 3 cases of death,2 patients died due to failure in recanalization,another patient died of severe pulmonary infection. Conclusion: The results suggest that this technique of a tri-axial system used of the Solitaire AB stent through a Navien delivery catheter can effectively retrieve clots from the occlusive artery and improves functional outcome in patients with acute ischemic stroke of vertebrobasilar artery.

Aclidinium bromide inhibits human glioma cell proliferation, migration and invasion and promotes apoptosis via the PI3K/AKT signaling pathway.

This study investigates the anti-cancer potential of Aclidinium bromide (INN) in glioblastoma. Glioblastoma cell lines U251 and U87 were treated with INN and its effects on cell migration and invasion were assessed by transwell migration and invasion assays., The effects of INN on proliferation and apoptosis were detected by CCK-8 kit and flow cytometry, and Western blotting determined anti-apoptotic proteins and signaling pathway changes. The results show that INN effectively suppressed proliferation, migration and invasion and induced apoptosis in U251 and U87 cells, respectively. Furthermore, the expression levels of the Bcl-2 anti-apoptotic protein was significantly decreased while Bax and caspase-3 expression were both increased in glioblastoma cells (all, p<0.05). Moreover, INN inactivated the PI3K/AKT signaling pathway by down-regulating the level of p-AKT, p-mTOR, P70 and CyclinD1 (all, p<0.05). In conclusion, our data suggests that INN could provide novel anticancer therapy in the treatment of glioblastoma.

Mechanism of lncRNA DUXAP8 in promoting proliferation of bladder cancer cells by regulating PTEN.

OBJECTIVE: To evaluate the lncRNA DUXAP8 expression in bladder cancer and its mechanism. PATIENTS AND METHODS: Clinical specimens were analyzed. The expression of lncRNA in bladder cancer and adjacent tissues was detected using qRT-PCR. The χ2-test analysis was used to analyze the relationship between lncRNA DUXAP8 expression and clinicopathological information in patients with bladder cancer. The tumor cell activity and cell proliferation were measured by cell counting kit-8 (CCK8) and colony formation assay. We utilized polymerase chain reaction (PCR) to access PTEN expression in bladder cancer and adjacent tissues. Pearson correlation analysis was utilized for evaluating the relationship between PTEN and lncRNA DUXAP8. Western blot was used for detecting protein expression. RESULTS: LncRNA DUXAP8 expression was higher in bladder cancer tissues; it was in a positive correlation with the TNM stage and tumor size, but negatively correlated with the total survival time. Knockdown of DUXAP8 decreased cell viability and cellular proliferation. Lower expression of PTEN gene was found in bladder cancer compared with that in adjacent tissues. Pearson correlation analysis showed that PTEN was negatively correlated with DUXAP8; knockdown of DUAP8 increased the expression of PTEN. Overexpressing DUAP8 increased protein level of PTEN, but decreased cell viability. CONCLUSIONS: Our results pointed out that lncRNA DUXAP8 was overexpressed in bladder cancer tissues, which can promote the progression of bladder cancer through inhibiting PTEN.

Value of texture analysis based on enhanced MRI for predicting an early therapeutic response to transcatheter arterial chemoembolisation combined with high-intensity focused ultrasound treatment in hepatocellular carcinoma.

AIM: To evaluate the potential value of texture analysis (TA) based on contrast-enhanced magnetic resonance imaging (MRI) for predicting an early response of patients with hepatocellular carcinoma (HCC) who were treated with transcatheter arterial chemoembolisation (TACE) combined with high-intensity focused ultrasound (HIFU). MATERIALS AND METHODS: Patients with HCC (n=89) who underwent contrast-enhanced MRI at 1.5 T 1 week before and 1 week, 1 month, and 3 months after TACE/HIFU were included in this retrospective study. Early responses were evaluated by two radiologists according to the Response Evaluation Criteria in Cancer of the Liver (RECICL). An independent Student's t-test and the Mann-Whitney U-test were used to compare the TA parameters between the complete response (CR) group and the non-complete response (NCR) group. Logistic regression and receiver operating characteristic (ROC) curve analyses were performed to assess the predictive value of the NCR lesions. RESULTS: Among the 89 patients, 58 showed CR and 31 showed NCR. Before TACE/HIFU, the CR group showed higher uniformity and energy but lower entropy than the NCR group (p<0.05). After TACE/HIFU, the CR group showed higher uniformity and energy but lower entropy and skewness than the NCR group (p<0.05). The logistic regression and ROC curve analyses showed that the entropy before TACE/HIFU and the skewness and entropy 1 week after TACE/HIFU were predictors of an early response. CONCLUSION: TA parameters based on contrast-enhanced MRI images 1 week before and after TACE/HIFU may act as imaging biomarkers to predict an early response of patients with HCC.

Detection of hand-foot-mouth disease and its spatial-temporal Epidemiological characteristics with GIS platform.

The aim of this study is to investigate the main category of hand-foot-mouth (HFM) virus and analyze the distribution characteristics and susceptible population of HFM disease in China. Infants who have had HFM disease for less than 7 days were selected from the First Affiliated Hospital of Guangxi Medical University, Guangxi, China. Various specimens were collected from the infants, and EV71 and CA16 nucleic acid detections were performed using fluorescence quantitative assay. The positive results of the specimens were compared to determine the components of the pathogen. Moreover, the data of the target cases were analyzed based on Geographic Information System (GIS) to obtain the spatial-temporal epidemiological features of HFM disease in China. The detection rate of HFM virus in the throat swab, feces, bleb fluid and cerebrospinal fluid were 75%, 81.13%, 85.71% and 25%, respectively, indicating that the detection rate of virus in the bleb fluid was the highest. When the detection was based on more than one specimen, it was found that the positive rate was higher compared to detection based on a single specimen. The positive detection rate of EV71 in the target specimens was significantly higher than that of CA16 and mixed infection. Moreover, CA16 infection was usually accompanied by EV71 infection. As to spatial-temporal distribution, hand-foot-month disease broke out in the South of China in April, then spread to the north, and diminished in July. There was a notable difference in the number of cases between different provinces. EV71 and CA16 are the main viruses inducing HFM disease, especially EV71. Fluorescence quantitative polymerase chain reaction with high sensitivity can be used to detect the copy number of viruses, which is applicable to the early diagnosis of HFM disease. The incidence of HFM disease is notably different according to the influence of time, geographical space, gender and the living conditions of the children. Early diagnosis and treatment based on scientific methods are needed to reduce the incidence of severe diseases and avoid death.

Risk factors associated with mortality from vascular thromboembolic events in patients diagnosed with non-small cell lung cancer: a population-based analysis.

BACKGROUND: Lung cancer patients are at increased risk for vascular events possibly due to cancer induced hypercoagulation. AIM: The purpose of this study was to evaluate risk factors associated with the mortality from vascular thromboembolic events in patients diagnosed with non-small cell lung cancer (NSCLC). DESIGN: Retrospective population-based analysis. METHODS: We used Surveillance, Epidemiology and End Results Program for 2004-13 and evaluated 199 337 patients with NSCLC. Univariate and multivariate subdistribution hazard regression models were used to identify potential risk factors for mortality from vascular thromboembolic events. Stratification analysis against clinical stage was performed to determine if the severity of the disease influenced the identified associations. RESULTS: Multivariate Cox regression analysis demonstrated that increased risk of mortality due to vascular thromboembolic events was associated with age, black race, non-adenocarcinoma histology, surgical treatment alone (all, P < 0.001) and north central region of SEER registry (P = 0.003). Female gender (P < 0.001), Asian or Pacific Islander race (P = 0.001), multiple co-existing primary cancers and late cancer stages (both, P < 0.001) were associated with significantly lower risk of mortality due to vascular thromboembolic events. The significant predictors of mortality from the vascular thromboembolic event were dependent on the stages of the disease. CONCLUSIONS: Risk factors associated with mortality from the vascular thromboembolic events in NSCLC patients identified in this study can promote awareness and may help to identify groups of patients that can benefit from anti-thrombotic prophylaxis measures.

Analysis of hemorrhagic fever with renal syndrome and its pathogenic gene sequence based on geographic information system.

This study analyzed the temporal-spatial distribution characteristics, epidemiological characteristics and gene sequences of hemorrhagic fever with renal syndrome (HFRS) in Guangxi, with the intention of providing a theoretical and technical support for the prevention of HFRS. A map of the incidence of HFRS of different cities in Guangxi was drawn up using the Geographic Information System (GIS) to investigate the epidemiological characteristics and infection source of HFRS between 2013 and 2016. Guangxi has a low incidence of HFRS, and autumn and winter are the main high-incidence seasons. Cases of HFRS were reported in all regions in Guangxi except Laibin city between 2013 and 2016. The distribution of cases in the four years suggested that Guilin, Nanning, Hechi and Wuzhou were the main infected regions, especially the local areas in the north of Guilin. The nucleotide and amino acid of S fragment and M fragment of Hantaviruses (HV) detected were highly homologous, and no obvious variation was found. Through analyzing the space-time characteristics, epidemiological characteristics and gene sequence of HFRS in Guangxi, it was found that areas rich in water, grass and moisture, such as paddy fields, are the main active areas for the host of HFRS.

Screening and identification of a tumor specific methylation phenotype in the colorectal laterally spreading tumor.

OBJECTIVE: We screened and identified the differential expression of the methylation phenotype in the whole genome of colorectal laterally spreading tumor (LSTs). MATERIALS AND METHODS: 3 tissue samples of colorectal polypoid adenomas (PAs), 3 tissue samples of LSTs and 3 tissue samples of colon cancer were analyzed with a high-density gene chip, and about 450,000 methylation sites were detected covering approximately 95% of the CpG islands. The Delta Data screening was taken through a cluster analysis of methylation phenotype differential expression. 50 tissue samples each of PAs patients, LSTs patients, and colorectal cancer patients were selected. Methylation-specific PCR (MSP) was used to detect RASSF1A and WIF-1 methylation levels. He RT-PCR method was used to detect the relative mRNA expression levels for methylation expression identification. RESULTS: The degree of LST methylation was higher than that of PAs, and 1234 genes were found to have a lower expression when compared to colorectal cancer samples. 764 genes had a higher expression when compared to colorectal cancer, and 559 genes lower expression when compared to PAs. The average methylation level of LSTs was higher than that of PAs, and lower than that of colorectal cancer. The chromosomal location was taken on these 1234 genes, which were higher than that of PAs, and lower than that of colorectal cancer; 518 genes were located on chromosome No. 2 (41.98%), 236 on No. 5 (19.12%), 357 on No. 8 (28.93%), and 123 on No. 10 (9.97%). According to clustering analysis, DNA differentially methylated sites were mainly on genes of cell adhesion molecules regulation, signaling pathways, energy transduction, cell cycle and apoptosis. The positive rate of RASSF1A and WIF-1 methylation in the tissues of LSTs patients were higher than that of PAs, and lower than that of colorectal cancer; differences were statistically significant (p<0.05). The relative expression levels of RASSF1A and WIF-1mRNA in the tissues of LSTs patients were lower than that of PAs, higher than that of colorectal cancer, and the difference was statistically significant (p<0.05). CONCLUSIONS: The administration of high-density gene chip technology has a good application value to screen the differential expression of LSTs gene methylation phenotype. Results are consistent with the identification results.

Functional calcium imaging in zebrafish lateral-line hair cells.

Sensory hair-cell development, function, and regeneration are fundamental processes that are challenging to study in mammalian systems. Zebrafish are an excellent alternative model to study hair cells because they have an external auxiliary organ called the lateral line. The hair cells of the lateral line are easily accessible, which makes them suitable for live, function-based fluorescence imaging. In this chapter, we describe methods to perform functional calcium imaging in zebrafish lateral-line hair cells. We compare genetically encoded calcium indicators that have been used previously to measure calcium in lateral-line hair cells. We also outline equipment required for calcium imaging and compare different imaging systems. Lastly, we discuss how to set up optimal imaging parameters and how to process and visualize calcium signals. Overall, using these methods, in vivo calcium imaging is a powerful tool to examine sensory hair-cell function in an intact organism.

Inhibition of subcutaneously implanted human pituitary tumor cells in nude mice by LRIG1.

The aim of this study was to explore the inhibition of subcutaneously implanted human pituitary tumor cells in nude mice by LRIG1 and its mechanism. For this study, athymic nude mice were injected with either normal pituitary tumor RC-4B/C cells or LRIG1-transfected RC-4B/C cells. We then calculated the volume inhibition rate of the tumors, as well as the apoptosis index of tumor cells and the expression of Ras, Raf, AKt, and ERK mRNA in tumor cells. Tumor cell morphological and structural changes were also observed under electron microscope. Our data showed that subcutaneous tumor growth was slowed or even halted in LRIG1-transfected tumors. The tumor volumes were significantly different between the two groups of mice (χ2 = 2.14, P < 0.05). The tumor apoptosis index was found to be 8.72% in the control group and 39.7% in LRIG1-transfected mice (χ2 = 7.59, P < 0.05). The levels of Ras, Raf, and AKt mRNA in LRIG1-transfected RC-4B/C cells were significantly reduced after transfection (P < 0.01). Transfected subcutaneous tumor cells appeared to be in early or late apoptosis under an electron microscope, while only a few subcutaneous tumor cells appeared to be undergoing apoptosis in the control group. In conclusion, the LRIG1 gene is able to inhibit proliferation and promote apoptosis in subcutaneously implanted human pituitary tumors in nude mice. The mechanism of LRIG1 may involve the inhibition of the PI3K/ Akt and Ras/Raf/ERK signal transduction pathways.

Ex vivo expansion of antigen-specific CD4+CD25+ regulatory T cells from autologous naïve CD4+ T cells of multiple sclerosis patients as a potential therapeutic approach.

OBJECTIVE: CD4+CD25+ regulatory T cells (Tregs) have been found to have a decreased effector function in patients with multiple sclerosis (MS). In this study, we co-cultured naïve CD4+ T cells of MS patients with myelin basic protein (MBP)85-99 peptide as specific antigen and allogenic B cells as antigen-presenting cells, in an attempt to generate adequate antigen-specific CD4+CD25+ Tregs with normal or improved immune function. PATIENTS AND METHODS: Naïve CD4+ T cells were isolated from peripheral blood mononuclear cells (PBMCs) from patients with MS (n=5) and healthy controls (HC, n=5). Furthermore, these purified naive CD4+ T cells were co-cultured with the CD40-activated B cells and MBP85-99 peptide to induce MBP-reactive CD4+CD25highCD127low Tregs. After harvesting these Tregs via a flow sorter, real-time PCR and mixed lymphocyte reaction (MLR) assay were performed to characterize cellular immune function. Supernatant interleukin (IL)-10 and transforming growth factor (TGF)-ß1 protein levels were detected by an enzyme-linked immunosorbent assay (ELISA). RESULTS: With this method, the frequency of CD4+CD25highCD127low Tregs in CD4+ T cells was 3.5%-6%. In both MS and HC groups, there were relatively lower proliferation indices (PI) of MLR assay but higher supernatant IL-10 and TGF-ß1 levels in the presence of MBP than those in the presence of other control antigens, where no significant differences were found. CONCLUSIONS: Via the ex vivo culture, adequate MBP-reactive CD4+CD25+ Tregsderived from autologous naïve CD4+ T cells of MS patients, were obtained and returned to normal without immune defects, and even upregulated their immunosuppressive function mostly through the elevated release of IL-10 and TGF-ß1.

Association of human leukocyte antigen DRB1 polymorphism and tuberculosis: a meta-analysis.

OBJECTIVE: To determine whether the human leukocyte antigen DRB1 (HLA-DRB1) is associated with clinical tuberculosis (TB). METHODS: Pooled odds ratios (OR) with 95% confidence intervals (CIs) were used to evaluate the strength of the association between HLA-DRB1 alleles and risk of TB. The χ(2)-based Q-test and I(2) statistics were calculated to examine heterogeneity. Egger's test was performed for the assessment of publication bias. Subgroup analysis was performed based on ethnicity and genotyping methods. RESULTS: A total of 19 case-control studies with 16 alleles (HLA-DRB1*01-HLA-DRB1*16) were included in this meta-analysis. No significant publication bias was detected among these studies. The HLA-DRB1*03 (OR 0.77, 95%CI 0.64-0.93, P = 0.0057) showed a protective effect, while HLA-DRB1*04 (OR 1.24, 95%CI 1.00-1.55, P = 0.0494), HLA-DRB1*08 (OR 1.45, 95%CI 1.14-1.86, P = 0.0030) and HLA-DRB1*16 (OR 1.39, 95%CI 1.04-1.87, P = 0.0269) were significantly associated with increased TB occurrence. Subgroup analysis showed that both ethnicity and genotyping method affected the association between HLA-DRB1*03, HLA-DRB1*04 and HLA-DRB1*08 alleles and TB occurrence. CONCLUSION: These results reinforce the importance of HLA-DRB1 alleles in the development of infectious diseases.

Long-term inhibition of Rho-kinase restores the LTP impaired in chronic forebrain ischemia rats by regulating GABAA and GABAB receptors.

We previously demonstrated that inactivation of Rho-kinase by hydroxyfasudil could impact N-methyl-d-aspartate (NMDA) excitatory interneurons in the hippocampus and attenuate the spatial learning and memory dysfunction of rats caused by chronic forebrain hypoperfusion ischemia. Complementary interactions between the excitatory neurotransmitter glutamate and the inhibitory neurotransmitter GABA form the molecular basis of synaptic plasticity and cognitive performance. However, whether the GABAergic inhibitory interneurons are involved in the mechanisms underlying these processes remains unclear. Here, we further examined the role of GABAergic interneurons in the neuroprotective effect of the Rho-kinase inhibitor. Chronic forebrain ischemia was induced in Wistar rats by bilateral common carotid artery occlusion (BCAO). The general synaptic transmission and long-term potentiation (LTP) of hippocampal CA3 neurons were evaluated at 30 days after sham surgery or BCAO. Real-time PCR and Western blot analyses were conducted to determine the effect of the Rho-kinase inhibitor hydroxyfasudil on GABAergic inhibitory interneuron expression and function after ischemia. Hydroxyfasudil showed no significant effect on general synaptic transmission, but it could abolish the inhibition of LTP induced by chronic forebrain ischemia. Moreover, the mRNA and protein levels of GABAA and GABAB in three brain regions after ischemia were markedly decreased, and hydroxyfasudil could up-regulate all mRNA and protein expression levels in these areas except for GABAA mRNA in the cerebral cortex and striatum. Using phosphorylation antibodies against specific sites on the GABAA and GABAB receptors, we further demonstrated that hydroxyfasudil could inhibit GABAergic interneuron phosphorylation triggered by the theta burst stimulation. In summary, our results indicated that the inactivation of Rho-kinase could enhance GABAA and GABAB expressions by different mechanisms to guarantee the induction of hippocampal LTP, and it could decrease the phosphorylation level of GABAergic inhibitory interneurons to promote the LTP induction rate and magnitude, hence improving the cognitive deficit suffered after chronic forebrain ischemia.