Circ-Slain2 Alleviates Cartilage Degradation and Inflammation of TMJOA.

Temporomandibular joint osteoarthritis (TMJOA) is a degenerative disease with the cessation of matrix anabolism and aggravation of inflammation, which results in severe pain and impaired joint function. However, the mechanisms are not well understood. Circular RNAs (circRNAs) are reported to have various biological functions and participate in the development, diagnosis, prognosis, and treatment of different diseases. This study aimed to investigate the roles and mechanisms of circ-slain2 in TMJOA. We first established TMJOA mouse models and found circ-slain2 was lowly expressed in the cartilage of TMJOA through sequencing data. We observed that circ-slain2 is predominantly localized in the cytoplasm and downregulated in mouse condylar chondrocytes (mCCs) treated with tumor necrosis factor α (TNFα) and interferon γ (IFNγ). Micro-computed tomography and histological examination showed that intra-articular injection of circ-slain2 overexpressing adeno-associated virus could alleviate cartilage catabolism and synovial inflammation to relieve TMJOA in vivo. In addition, elevated circ-slain2 also showed anticatabolic and anti-inflammatory effects on IFNγ- and TNFα-stimulated mouse condylar chondrocytes (mCCs). Functional enrichment analysis indicated that protein processing in endoplasmic reticulum (ER) was associated with TMJOA, and further functional experiments confirmed that circ-slain2 could suppress ER stress in OA mCCs. RNA binding protein immunoprecipitation assay revealed an overt interaction between activating transcription factor 6 (ATF6) and circ-slain2. Inhibition of the expression of both ATF6 and circ-slain2 resulted in dilation of the ER and enhanced the expression of ER stress markers, whose ER stress level was higher than inhibition of ATF6 but lower than knockdown of circ-slain2 expression. Collectively, our research demonstrated that circ-slain2 could regulate ATF6 to relieve ER stress, reducing temporomandibular joint cartilage degradation and synovial inflammation. These findings provide prospects for developing novel osteoarthritis therapies based on circ-slain2 by focusing on reducing the inflammation of synovium and the imbalance between matrix synthesis and degradation.

The prognostic value of circulating tumor DNA in patients with melanoma: A systematic review and meta-analysis.

BACKGROUND: Circulating tumor DNA (ctDNA) has been investigated as a potential prognostic biomarker to evaluate the therapeutic efficacy and disease progression in melanoma patients, yet results remain inconclusive. The purpose of this study was to illustrate the prognostic value of ctDNA in melanoma. OBJECTIVES: To describe the clinical prognostic value of ctDNA for melanoma patients. METHODS: Searched for eligible articles from Pubmed, Web of Science and Embase. Pooled hazard ratios (HRs) and 95% confidence intervals (CIs) were calculated to evaluate the association between ctDNA at baseline or during treatment and overall survival (OS) and progression-free survival (PFS). RESULTS: A total of 9 articles were obtained, involving 617 melanoma patients. The pooled HRs revealed that compared with baseline undetectable ctDNA patients, detectable ctDNA was highly correlated with poor OS (HR 2.91, 95% CI: 2.22-3.82; p < 0.001) and PFS (HR 2.75, 95% CI: 1.98-3.83; p < 0.001). A meta-analysis of these adjusted HRs was performed and confirmed that ctDNA collected at baseline was associated with poorer OS/PFS (OS: HR 3.00, 95% CI 2.19-4.11, p < 0.001/PFS: HR 2.68, 95% CI 1.77-4.06, p < 0.001). During treatment, a significant association was shown between ctDNA and poorer OS/PFS (OS: HR 6.26, 95% CI 2.48-15.80, p < 0.001; PFS: HR 4.93, 95% CI 2.36-10.33, p < 0.001). CONCLUSION: Investigation and application of ctDNA will improve "liquid biopsy" and play a role in early prediction, monitoring disease progression and precise adjusting treatment strategies in melanoma patients.

A 12-week subchronic intramuscular toxicity study of risperidone-loaded microspheres in rats.

Long-acting injectable formulations of antipsychotics have been an important treatment option to increase the compliance of the patient with schizophrenia by monitoring drug administration and identifying medication noncompliance and to improve the long-term management of schizophrenia. Risperidone, a serotoninergic 5-HT2 and dopaminergic D2 receptor antagonist, was developed to be a long-acting sustained-release formulation for the treatment of schizophrenia. In this study, 12-week subchronic toxicity study of risperidone-loaded microspheres (RMs) in rats by intramuscular injection with an 8-week recovery phase was carried out to investigate the potential subchronic toxicity of a novel long-acting sustained-release formulation. The results indicated that the dosage of 10-90 mg/kg of RM for 2 weeks did not cause treatment-related mortality. The main drug-related findings were contributed to the dopamine D2 receptor and α1-adrenoceptor antagonism of risperidone such as elevation of serum and pituitary prolactin levels and ptosis and changes in reproductive system (uterus, ovary, vagina, mammary gland, testis, seminal vesicle, epididymis, and prostate). In addition, foreign body granuloma in muscle at injection sites caused by poly-lactide-co-glycolide was observed. At the end of the recovery phase, these changes mostly returned to normal. The results indicated that RM had a good safety profile in rats.

A 12-week intramuscular toxicity study of risperidone-loaded microspheres in Beagle dogs.

Long-acting formulations of antipsychotics are important treatment options to increase the compliance of schizophrenic patients. Risperidone, a 5-HT2 and dopaminergic D2 receptor antagonist, was developed as long-acting sustained-release microspheres with poly(lactide-co-glycolide) (PLGA) as a drug carrier for the treatment of schizophrenia. In the present study, the main objective is to determine the nonclinical safety profile of risperidone-loaded microspheres (RM) in Beagle dogs after intramuscular administration for 3 months, once in 2 weeks, followed by 8-week recovery phase. No animal death was found and no special toxicological findings were observed. The findings, such as hypoactivity, ptosis, increased heart rate, and elevated serum and pituitary prolactin levels, were observed and related to the pharmacological effects of risperidone. The changes in the reproductive system (uterus, ovary, vagina, cervix, and mammary gland) were considered secondary to the prolactin elevation, and the congestion of spleen was related to risperidone. The foreign body granulomas at injection sites might be caused by PLGA. At the end of recovery phase, the above changes mostly recovered to normal, and on administering 3 mg/kg dose level once in 2 weeks on Beagle dogs showed no observed adverse effect. Taken together, RM had exhibited the acceptable safety.

TRBV kinetics and its association with HLA disparity and aGVHD following allogeneic hematopoietic stem cell transplantation.

INTRODUCTION: The relative expression of T cell receptor (TCR) beta variable (TRBV) and TCR diversity was compared between recipients receiving human leukocyte antigen (HLA)-mismatched transplants and those receiving HLA-matched transplants, using granulocyte colony-stimulating factor (G-CSF)-mobilized peripheral blood stem cells and bone marrow as grafts. METHODS: The kinetics of the relative expression of TRBV family members were analyzed using real-time quantitative PCR. Additionally, the association of TRBV clonotype with acute graft-versus-host disease (aGVHD) was determined by cloning and sequence analysis of the CDR3 region. RESULTS: The TCR diversity in recipients receiving HLA-mismatched transplants was significantly lower than in those receiving HLA-matched transplants at 1 month and 2 months after hematopoietic stem cell transplantation (HSCT) (both P < 0.05). However, these differences disappeared 3 months after transplantation. The relative expression of TRBV27 (n = 7 recipients) at the onset of aGVHD was higher than in corresponding donors (P = 0.025), but no significant differences were observed between recipients lacking aGVHD and their donors at serial time points after HSCT. CONCLUSION: Our results suggest that HLA disparity may affect the relative expression of TRBV in the early phase after transplantation, and TRBV27 may be associated with the onset of aGVHD.

Identification of Piccolo as a regulator of behavioral plasticity and dopamine transporter internalization.

Dopamine transporter (DAT) internalization is a mechanism underlying the decreased dopamine reuptake caused by addictive drugs like methamphetamine (METH). We found that Piccolo, a presynaptic scaffolding protein, was overexpressed in the nucleus accumbens (NAc) of the mice repeatedly administrated with METH. Piccolo downexpression by antisense technique augmented METH-induced behavioral sensitization, conditioned reward and synaptic dopamine accumulation in NAc. Expression of Piccolo C2A domain attenuated METH-induced inhibition of dopamine uptake in PC12 cells expressing human DAT. Consistent with this, it slowed down the accelerated DAT internalization induced by METH, thus maintaining the presentation of plasmalemmal DAT. In immunostaining and structural modeling Piccolo C2A domain displays an unusual feature of sequestering membrane phosphatidylinositol 4,5-bisphosphate, which may underlie its role in modulating DAT internalization. Together, our results indicate that Piccolo upregulation induced by METH represents a homeostatic response in the NAc to excessive dopaminergic transmission. Piccolo C2A domain may act as a cytoskeletal regulator for plasmalemmal DAT internalization, which may underlie its contributions in behavioral plasticity.

[Large-capacity expanded cytoline-induced killer cells and its cytotoxic activity].

This study was conducted to establish the large-capacity culture methd of cytokine-induced killer (CIK) cells for clinical therapy and assess its effect on the fuction of cell-mediated immunity following autologous CIK cells reinfusion. Autologus CIK cells were expanded in 1000 ml culture-bag and reinfused back. The MTT method was used to test the cytotoxic activity of CIK cells before and after reinfusion. The results showed that the total amount of autologous CIK cells reinfusion exceeded 1.6 x 10(10) with the use of the culture method of large-capacity. The PBMNC from patients treated by CIK cells showed significant increase in cytotoxic activity, no side effects were observed, and therefore the large-capacity culture method of CIK cells is a simle and safe therapy for treating the minimum residue of diseases.

[Zinc modulates the Ca(2+)-ATPase activity of rat osteoblast membrane].

The effect and mechanism of zinc on Ca(2+)-ATPase and Na+, K(+)-ATPase activity of rat osteoblast membrane were studied. The activities were measured by colorimetric method. The effect of special inhibitors of protein kinase C or calmodulin on membrane Ca(2+)-ATPase and Na+, K(+)-ATPase activity which induced by zinc were also examined. The results showed that zinc could increase membrane Ca(2+)-ATPase activity, but no influence was observed on Na+, K(+)-ATPase activity. R24571, a special inhibitor of calmodulin, had no effect on membrane Ca(2+)-ATPase activity. Calmodulin also could not promote the basic Ca(2+)-ATPase activity of membrane. But trifluoperazine, a special inhibitor of protein kinase C, obviously reduced its activity. It was concluded that the modulation of zinc on membrane Ca(2+)-ATPase activity of rat osteoblast could be mediated by protein kinase C.

[Simultaneous assay of Ca(2+)-ATPase and Na+, K(+)-ATPase activities of osteoblast rat by malachite green colorimetic method].

The aim of this study was to find a method to measure simultaneously the activities of Ca(2+)-ATPase and Na+, K(+)-ATPase of rat osteoblast membrane. In this test, Ca(2+)-ATPase and Na+, K(+)-ATPase released inorganic phosphate (Pi) from ATP, and Pi was determined by colorimetric method using malachite green dye to show ATPase activities. Ca2+ concentration and osteoblast culture generation were important factors affecting ATPase activities. Ca2+ could elevate membrane ATPase activity. When Ca2+ concentration reached 40 mumol/L, ATPase activities were improved. Ca(2+)-ATPase activity decreased with the increase of osteoblast culture generation. Ca(2+)-ATPase activity was lower than Na+, K(+)-ATPase activity in osteoblast membrane. This method is simple, sensitive and reliable.

[A study on relationship between chemotherapy of acute leukemia and apoptosis in vivo].

In order to investigate whether apoptosis occurs in vivo in patients with leukemia in different stage (before, during and after chemotherapy). We detect apoptosis of peripheral blood leukemia cells in 10 patients before, during and after chemotherapy with in situ TdT fluorescence measurement and DNA electrophoresis. The results showed that apoptotic cells were few (0-2.9%) before chemotherapy, increased obviously (11.4%-72.0%) during chemotherapy in patients with complete remission (CR) or partial remission (PR), but decreased (0-2.9%) after chemotherapy (10-14 days). Apoptotic cells were few (0-5.7%) during chemotherapy in four patients without remission. Therefore, only effective drug therapy could induce apoptosis. DNA ladder cannot be detected by agarose gel electrophoresis either before, during and after chemotherapy. So we considered that in situ TdT assay is possible to apoptosis in vivo in the early course of chemotherapy for immediate modification of chemotherapy, while electrophoretic analysis is not sensitive enough to detect apoptotic cells in vivo.

[Study on the dose-effect relationship of hypolipidemic effect of deepsea fish oil].

The dose-effect relationship of hypolipidemic effect of fish oil was studied. The rats were divided into four groups: 2.5, 5, 10 g/kg and high dietary fat control group. The experimental rat model of high serum lipids was developed. The results showed that there was a linear correlation between the levels of TG, HDL-c and the dosage of fish oil. Fish oil reduced TC level and elevated HDL-c level steadily. But no linear correlation was observed between TC, LDL-c and the dosage of fish oil. The level of TC and LDL-c decreased markedly at the dosage of 2.5 g/kg of fish oil.