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.

Application of case-based learning in medical student education: a meta-analysis.

OBJECTIVE: To evaluate the effectiveness of case-based learning (CBL) in medical students' education through meta-analysis. MATERIALS AND METHODS: PubMed, Cochrane Library, Elsevier and other databases were searched to find randomized controlled trials (RCTs) of CBL teaching methods and other teaching methods published from January 1, 1995, to October 1, 2020. All included studies used the Cochrane risk bias assessment tool, and Review Manager software, version 5.3 (Copenhagen, Denmark), was used for the meta-analysis and systematic review. RESULTS: A total of 8 studies were included with a total of 939 students, including 480 in the CBL group and 459 in the control group. Compared with other teaching methods, CBL teaching can improve medical students' academic performance (p=0.03) and case analysis ability (p<0.001). CONCLUSIONS: CBL is an active teaching method that is effective for educating medical students and helps to improve their performance and case analysis ability.

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.

[Comparison of radiofrequency catheter ablation for paroxysmal supraventricular tachycardia guided by three dimensional navigation with X-ray].

Objective: To compare the therapeutic efficiency and safety of radiofrequency catheter ablation (RFCA) for paroxysmal ventricular tachycardia (PSVT) guided by three-dimensional navigation with X-ray. Methods: One hundred and seventy-six patients with PSVT hospitalized in Department of Cardiology, Zhejiang Provincial People's Hospital between January 2013 and December 2014 were enrolled in this study. RFCA was performed on 95 patients (Group A) guided by the three dimensional electric-field navigation system (NavX) and the procedures were done with the spatial localization method, unless X-ray was needed in some cases.Eighty-one patients (Group B) underwent RFCA guided by X-ray only.The success rate, complications, recurrence rate, operation time, and X-ray exposure were compared between the two groups. Results: The immediate success rate of operation was 100% in the 176 PSVT patients without complications.There were recurrences in 2 cases of Group A, while in 1 case of Group B. The average operation time in Group A was (97±20) min, while (91±26) min in Group B. The median X-ray fluoroscopy time was 3.0 min and radiation dose was 18.5 µGym2 in Group A, which were significantly reduced compared with those in Group B (34.5 min, 167.3 µGym2) (P<0.05). There was no significant difference in immediate procedure success rate, complications, recurrence, and operative time between this two groups (P>0.05). Conclusions: The RFCA for PSVT guided by three-dimensional navigation is safe and feasible in the experienced electrophysiological center.Most patients suffer less or no X-ray radiation.

Evaluation of gamma ray-induced gastrointestinal tract morphological and proliferative activity changes in rhesus monkeys.

To provide support for future pharmacology and preclinical studies, we have established a stable nonhuman primate animal model to demonstrate the histopathological changes in the gastrointestinal tract following gamma ray irradiation. In this study, 12 healthy rhesus monkeys were divided into 2 groups (control and radiation groups). Animals in the radiation group were exposed to gamma rays (cobalt 60 source) at a dose level of 6.5 Gy total body irradiation bilaterally (i.e. 3.25 Gy on each side). Control animals were sham exposed using identical procedures. After a 5-day in-life observation period, gastrointestinal tract tissues (esophagus, stomach, duodenum, jejunum, ileum, colon, and rectum) were collected and fixed in 10% neutral-buffered formalin for subsequent hematoxylin and eosin and 5-bromo-2-deoxyuridine (BrdU) immunohistochemistry processing. The results showed that the esophagus was undergoing degeneration without obvious inflammatory changes, while the stomach and duodenum exhibited both degeneration and inflammation. From the jejunum to the rectum, late-stage inflammation with glandular regeneration, as well as a high-level BrdU labeling index, was present.

High e+/e- Ratio Dense Pair Creation with 10(21)W.cm(-2) Laser Irradiating Solid Targets.

We report results of new pair creation experiments using ~100 Joule pulses of the Texas Petawatt Laser to irradiate solid gold and platinum targets, with intensities up to ~1.9 × 10(21) W.cm(-2) and pulse durations as short as ~130 fs. Positron to electron (e+/e-) ratios >15% were observed for many thick disk and rod targets, with the highest e+/e- ratio reaching ~50% for a Pt rod. The inferred pair yield was ~ few ×10(10) with emerging pair density reaching ~10(15)/cm(3) so that the pair skin depth becomes < pair jet transverse size. These results represent major milestones towards the goal of creating a significant quantity of dense pair-dominated plasmas with e+/e- approaching 100% and pair skin depth ≪ pair plasma size, which will have wide-ranging applications to astrophysics and fundamental physics.

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.

¹H NMR-based metabonomics in brain nucleus accumbens and striatum following repeated cocaine treatment in rats.

Studies have shown a few cerebral metabolites modified by cocaine in brain regions; however, endogenous metabolic profiling has been lacking. Ex vivo (1)H NMR (hydrogen-1 nuclear magnetic resonance) spectroscopy-based metabonomic approach coupled with partial least squares was applied to investigate the changes of cerebral metabolites in nucleus accumbens (NAc) and striatum of rats subjected to cocaine treatment. Our results showed that both single and repeated cocaine treatment can induce significant changes in a couple of cerebral metabolites. The increase of neurotransmitters glutamate and gamma-amino butyric acid (GABA) were observed in NAc and striatum from the rats repeatedly treated with cocaine. Creatine and taurine increased in NAc whereas taurine increased and creatine decreased in striatum after repeated cocaine treatment. Elevation of N-acetylaspartate in NAc and striatum and decrease of lactate in striatum were observed, which may reflect the mitochondria dysregulation caused by cocaine; moreover, alterations of choline, phosphocholine and glycerol in NAc and striatum could be related to membrane disruption. Moreover, groups of rats with and without conditioned place preference (CPP) apparatus are presenting difference in metabolites. Collectively, our results provide the first evidence of metabonomic profiling of NAc and striatum in response to cocaine, exhibiting a regionally-specific alteration patterns. We find that repeated cocaine administration leads to significant metabolite alterations, which are involved in neurotransmitter disturbance, oxidative stress, mitochondria dysregulation and membrane disruption in brain.

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.

Noradrenaline in the bed nucleus of the stria terminalis is critical for stress-induced reactivation of morphine-conditioned place preference in rats.

The effect of noradrenaline in the bed nucleus of the stria terminalis and locus coeruleus on maintenance and reactivation of morphine-conditioned place preference induced by footshock stress was investigated in rats. After receiving alternate injection of morphine (10 mg/kg) and saline for 6 consecutive days, the rats spent more time in the drug-paired compartment (morphine-conditioned place preference) on day 7. These animals did not show morphine-conditioned place preference on day 37 following sham-footshock once every 3 days from days 8 to 36 (28 days drug-free). However, 15 min of intermittent footshock once every 3 days could induce the maintenance of morphine-conditioned place preference on day 37 with significantly more time spent in the drug-paired compartment than on day 0. Microinjection of the alpha(2)-adrenoceptor agonist, clonidine (0.1 or 1 microg), into the locus coeruleus 30 min before footshock did not affect stress-induced maintenance of conditioned place preference. However, infusions of clonidine (1 microg) into the bed nucleus of the stria terminalis significantly attenuated the maintenance of conditioned place preference induced by footshock stress. In another experiment, after a 21-day extinction of morphine-conditioned place preference, a single footshock could reactivate the morphine place preference that was significantly blocked by pretreatment with infusion of clonidine (0.1 or 1 microg) into the bed nucleus of the stria terminalis but not the locus coeruleus. Reactivation of morphine-conditioned place preference elicited by footshock stress was significantly inhibited by 6-hydroxydopamine-induced lesions in the ventral noradrenergic bundle, most of the norepinephrine input to the bed nucleus of the stria terminalis arising from caudal brain stem noradrenergic cell groups. In contrast, chemical lesions of the dorsal noradrenergic bundle that arises from the locus coeruleus had no such effects. These findings suggest that noradrenergic neurons in locus coeruleus are not involved in stress-induced reinstatement of drug-seeking and further clearly demonstrate that noradrenaline in the bed nucleus of the stria terminalis plays a critical role in mediating this effect. Comprehension of the neurochemical events underlying the stress-induced and the bed nucleus of the stria terminalis-mediated reinstatement of drug-seeking may, therefore, throw more light on the biological bases of drug dependence and addictive behavior

[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.

[Role of metallothionein in the resistance of rat osteoblasts to zinc excess and deficiency].

This study is expected to investigate the role of metallothionein(MT) in the resistance of osteoblasts to high or low zinc conditions. Antisense MT oligonucleotide was used to reduce the content of MT in osteoblast. Cellular MT content was determined by silver/hemolysate saturation method, and the cytotoxicity was measured by the activity of LDH releasing from damaged cells. The results showed that antisense MT oligonucleotide markedly decreased the resistance of osteoblast to zinc excess and zinc deficiency. It suggested that MT played an important role in the modulation and maintenance of intracellular zinc homeostasis of osteoblasts.

[Prospects on the application of toxicogenomics in assessing the safety of chemical compounds].

Toxicogenomics has been developing with the rapid advances in Human Genome Project in recent two years. The categories and specialty of tox-gene; the application of tox-gene microarray in safety assessment, analysis of gene mutation and gene polymorphism, extrapolation of animal tests results, clarification of mechanism and safety assessment of mixtures; some problems of toxicogenomics need to be solved in spite of its great prospective application.

[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.

[Zinc deficiency inhibited the proliferation and differentiation of rat osteoblast in vitro].

The effect of zinc deficiency on proliferation and differentiation of rat osteoblast was studied in vitro. The osteoblasts were isolated from the rat calvaria, and were subcultured in DMEM medium. Zinc deficiency model in cell culture was made by chelating zinc from the medium with a special chelator TPEN. Incorporation of 3H-thymidine was measured to show the DNA synthesis at different time point, and cell cycle was analyzed by flow cytometry. A histochemistry method was used to detect changes of osteoblast cytoskeleton. Incorporation of 3H-proline was measured to show the synthesis of collagen. Alkaline phosphatase activity (AKP) and osteocalcin content were measured by enzyme dynamics method and radioimmunoassay respectively. The results showed that the incorporation of 3H-thymidine in the zinc deficient group was significantly less than that in the control group at different time points. The cells were blocked at G2/M phase of cell cycle, which is related to the impairment of cytoskeleton. Collagen synthesis, osteocalcin content and AKP activity in the zinc deficient group were reduced significantly as compared with those in the control group. But no differences in all the measurements were observed between the control group and the group of zinc deficiency supplemented with zinc. Therefore, it was concluded that zinc deficiency inhibited the proliferation and differentiation of rat osteoblast in vitro.

[Zinc promotes proliferation and differentiation of osteoblast in rats in vitro].

OBJECTIVE: Effect of zinc on proliferation and differentiation of osteoblast in rats was studied in vitro. METHODS: Osteoblasts were isolated from the rat calvaria and subcultured in DMEM media. Rats were divided into four groups in the study, with administration of 0 micromol/L (as control), 10 micromol/L, 25 micromol/L and 50 micromol/L of Zn(2+), respectively. DNA synthesis at varied time points was determined with incorporation of (3)H-thymidine, and cell cycle was analyzed by flow cytometry. Synthesis of collagen was measured with incorporation of (3)H-proline. Activity of alkaline phosphatase (AKP) and osteocalcin content were measured by enzyme dynamics method and radioimmunoassay, respectively. RESULTS: Incorporation of (3)H-thymidine in the three groups with different zinc-supplement was significantly higher than that in the control group at each time point, and supplement with 25 micromol/L and 50 micromol/L of Zn(2+) could promote the transition of cell cycle from phases G(0)/G(1) to S phase. Syntheses of collagen and osteocalcin and activity of AKP in the groups with zinc-supplement were greater, as compared with those in the control group. CONCLUSION: Zinc could promote proliferation and differentiation of the rat osteoblast in vitro.

Apoptosis induced by zinc deficiency in rat osteoblast: possible involvement of protein kinase C.

Rat osteoblasts were isolated from the 21-day fetal rat calvarias. The cells were grown in DMEM plus 10% FBS, and were treated for 24 h. with 10 mumol/L TPEN or 10 mumol/L TPEN supplemented with 10 mumol/L Zn2+. Apoptosis of osteoblasts were measured by flow cytometry, electron microscopy and DNA fragmentation analyzed by gel electrophoresis. In addition, IP3 production and PKC activity were measured in order to show whether they are involved in apoptosis in osteoblast induced by zinc deficiency. The results showed that 10 mumol/L TPEN could induce apoptosis in osteoblast in 24 h. But cells treated with 10 mumol/L TPEN supplemented with 10 mumol/L Zn2+ showed no apoptotic changes in 24 h. TPEN significantly reduced the formation of IP3 and PKC activity after 24 h incubation. No differences were observed between the cells treated with TPEN supplemented with Zn2+ simultaneously and the untreated cells. It can be inferred that apoptosis induced by zinc deficiency may be due to the decreased activity of PKC which is impaired by reduced formation of IP3.