Objective To evaluate the mitochondrial function of chondrocytes in Kashin-Beck disease (KBD) patients and its correlation with age.Methods Mitochondrial function was evaluated by analyzing the activities of respiratory chain enzyme complexes and citrate synthase (CS),intracellular adenosine triphosphate (ATP)contents, changes in mitochondrial membrane potential (ΔΨm),as well as the intracellular reactive oxygen species (ROS) contents.Correlation of mitodhondrial function and age was analyzed with linear regression.Results Activities of complexes Ⅱ (P = 0.001 ),Ⅲ (P = 0.005 )and Ⅳ (P = 0.005 )were reduced in KBD articular chondrocytes compared with cells from normal controls.However,the mitochondrial mass was increased in KBD samples. Cultured KBD chondrocytes had reduced cellular ATP level (P = 0.001 )and mitochondrial membrane potential (P =0.001),but increased citrate synthase activity (P =0.04)and intracellular ROS level (P <0.001).We found no correlation between mitochondrial function and donor age either in normal or in KBD chondrocytes. Conclusion Mitochondrial function is altered in articular chondrocytes of KBD,and no correlation was found between mitochondrial function and donor age.

Objective To compare the effects of selenium nanoparticles (Nano-Se) and sodium selenium (Na2SeO3) on apoptosis and reactive oxygen species (ROS) of articular chondrocytes from patients with Kashin-Beck Disease (KBD) in vitro,and provide a scientific basis for preventing KBD.Methods The subjects with KBD were diagnosed on National Clinical Diagnostic Criteria of KBD (WS/T207-2010),articular cartilage from 8 patients undertaken joint replacement operation were collected.In vitro,chondrocytes were treated with concentration of 0,25,50,100,200,300,400 and 500 μg/L of Nano-Se and Na2SeO3 for 5 d,respectively.Cell growth was detected by MTT assay,and the highest concentration and time corresponding to the highest survival rate of Nano-Se and Na2SeO3 were used in the following experiment.KBD chondrocytes were treated with Nano-Se and Na2SeO3,and divided into control group,Na2SeO3 group,Nano-Se group according to the randomized design.Each group had 8 cases.The cell apoptosis and ROS were detected by flow cytometry.Results The optimal intervention concentration of Nano-Se and Na2SeO3 was 100 and 400 μg/L,respectively.The optimal intervention time of NanoSe and Na2SeO3 both was 3 days.There was a significant decrease in the total and terminal apoptosis,ROS level of chondrocytes in Nano-Se group [(4.67 ± 0.89)％,(1.51 ± 0.48)％,(56.04 ± 4.81)％] and Na2SeO3 group [(7.07 ±0.25)％,(4.37 ± 0.37)％,(87.13 ± 6.60)％] compared with those of control group [(9.95 ± 0.38)％,(6.93 ± 0.42)％,(125.17 ± 16.60)％,all P ＜ 0.01].The difference of early apoptotic rate among control group,Na2SeO3 group,NanoSe group [(3.02 ± 0.41)％,(2.7 ± 0.46)％,(3.16 ± 0.56)％] was not statistically significant (F =2.11,P =0.35).Conclusion Appropriate concentration of Nano-Se can significantly decrease oxidative stress of KBD chondrocytes and inhibit apoptosis compared to Na2SeO3.

Objective To explore convenient and practical semi-quantitative MRS indicators in diagnosis of prostate cancer.Methods One hundred patients with completer MRS data and clinical data were enrolled in the study.The following parameters were assessed to evaluate their efficacy in diagnosis of prostate cancer, (Cho + Cr)/Cit of single voxel, mean (Cho + Cr)/Cit of the whole area and (Cho + Cr)/ Cit positive voxel ratio at the sextant level.Similarly, all the parameters mentioned above and mean (Cho +Cr)/Cit positive voxel ratios of the whole gland were assessed at the whole gland level.Pearson test and Kappa test were used in the research.Results Two thousand nine hundred and forty-five voxels were assessed including 1203 in cancer positive region and 1742 in cancer negative regioa The range of the (Cho + Cr) /Cit ratio in cancer positive region was from 0.22 to 8.00 (median, 1.87), and that in cancer negative region was from 0.11 to 8.00 (median, 0.53).The (Cho + Cr)/Cit ratio in cancer positive region was higher than that in cancer negative region (Z =28.48, P<0.01) with partial overlap.On the level of sextant, the sensitivity, specificity, positive predictive value, negative predictive value and accuracy of (Cho + Cr)/Cit ratio> 0.911 to diagnose prostate cancer were 81.4% (179/220),64.2% (194/302), 62.4% (179/287) 、82.6% (194/235) 、71.5% (373/522) respectively; those of mean (Cho + Cr)/Cit ratio>0.911 were 77.3% (170/220),77.2% (233/302),71.1% (170/239),82.3% (233/283),77.2% (403/522) respectively; those of positive voxel ratio>0.519 were 73.2% (161/220),80.8% (244/302)、 73.5% (161/219),80.8% (244/302), 77.6% (405/522) respectively; the consistency between mean (Cho + Cr) /Cit ratio and positive voxel ratio was high (Kappa=0.907).On the level of the whole prostate, the sensitivity, specificity, positive predictive value, negative predictive value and accuracy of (Cho + Cr)/ Cit ratio>0.911 to diagnose prostate cancer were 94.3% (50/53),40.4% (19/47),64.1% (50/78), 86.4% (19/22),69.0% (69/100) respectively; those of mean (Cho + Cr)/Cit ratio>0.911 were73.6% (39/53),83.0% (39/47), 83.0% (39/47), 73.6% (39/53), 78.0% (78/100) respectively; those of maximum positive voxel ratio> 0.519 were 88.7% (47/53),61.7% (29/47),72.3% (47/65),82.9% (29/35),76.0% (76/100) respectively; those of mean positive voxel ratio>0.519 were 62.3% (33/53), 85.1% (40/47), 82.5% (33/40), 66.7% (40/60), 73.0% (73/100) respectively; the consistency between mean (Cho + Cr)/Cit ratio and mean positive voxel ratio was fairly high (Kappa =0.818).Conclusion Single voxel criteria were suggested to diagnose clinically suspected prostate cancer.Maximum positive voxel ratio criteria were suggested to guide localization in biopsy.

Objective: To investigate the pyroptosis-inducing effects of celastrol on tumor cells and to explore the potential mechanisms involved, specifically focusing on the role of the caspase-3/gasdermin E (GSDME) signaling pathway and the impact of endoplasmic reticulum (ER) stress and autophagy. Methods: Necrostatin-1 (Nec-1), lactate dehydrogenase release (LDH) assay, and Hoechst/propidium iodide (PI) double staining were employed to validate the mode of cell death. Western blot was used to detect the cleavage of GSDME and the expression of light chain 3 (LC3) and BIP. Results: Celastrol induced cell swelling with large bubbles, which is consistent with the pyroptotic phenotype. Moreover, treatment with celastrol induced GSDME cleavage, indicating the activation of GSDME-mediated pyroptosis. GSDME knockout via CRISPR/Cas9 blocked the pyroptotic morphology of celastrol in HeLa cells. In addition, cleavage of GSDME was attenuated by a specific caspase-3 inhibitor in celastrol-treated cells, suggesting that GSDME activation was induced by caspase-3. Mechanistically, celastrol induced endoplasmic reticulum (ER) stress and autophagy in HeLa cells, and other ER stress inducers produced effects consistent with those of celastrol. Conclusion These findings suggest that celastrol triggers caspase-3/GSDME-dependent pyroptosis via activation of ER stress, which may shed light on the potential antitumor clinical applications of celastrol.

OBJECTIVETo observe the effect of nano-Se-chondroitin sulfate on the growth and apoptosis of chondrocytes from patients with Kashin-Beck disease (KBD) exposed to T-2 toxin in vitro.

METHODSSamples of the articular cartilage were obtained from 6 patients with grade II/III KBD diagnosed in line with the National Clinical Diagnostic Criteria of KBD (WS/T 207-2010) for chondrocyte separation and culture in vitro. The separated chondrocytes were treated with synthesized nano-Se-chondroitin sulfate particles and T-2 toxin, alone or in combination, and the cell growth and apoptosis were observed using MTT assay, HE staining and flow cytometry.

RESULTSThe synthesized nano-Se-chondroitin sulfate, with a selenium entrapment ratio of 10.1%, spontaneously formed nanoparticles in distilled water with sizes ranging from 30 to 200 nm. Fourier-transform infrared spectroscopy suggested a possible covalent bond that bound Nano-Se and chondroitin sulfate. Within the concentration range of 50-200 ng/ml, nano-Se-chondroitin sulfate significantly inhibited T-2 toxin-induced apoptosis of the cultured chondrocytes and reduced the early apoptosis rate to (8.64∓1.57)% (P<0.05).

CONCLUSIONNano-Se-chondroitin sulfate can inhibit T-2 toxin-induced apoptosis of cultured chondrocytes from KBD patients in vitro, and serves as a promising candidate therapeutic agent for KBD.

Apoptosis ; drug effects ; Cells, Cultured ; Chondrocytes ; drug effects ; pathology ; Chondroitin Sulfates ; administration & dosage ; pharmacology ; Humans ; Kashin-Beck Disease ; pathology ; Middle Aged ; Nanostructures ; T-2 Toxin ; toxicity

This study was aimed to observe the effects on glucolipid metabolism of aqueous extract of traditional South African herb Sutherlandia. Intraperitoneal injection of streptozotocin (STZ) combined with high fat feed method was used in the establishment of type 2 diabetes rat model. Then, rats were randomly divided into the normal group, model group, pioglitazone group, and the Sutherlandia group. Observation was made on changes of body weight, oral glucose tolerance test (OGTT), blood triglycerides (TG), total cholesterol (TC), low density lipoprotein (LDL-C), high density lipoprotein (HDL-C). Western blot method was used to detect IRS-1 expression of skeletal muscle in rats of each group. The results showed that compared with the normal group, body weight in the model group was decreased, and indicators of OGTT, TG, TC, LDL-C were significantly increased (P< 0.05). Symptoms such as increased drink-ing, eating and urine were obvious; and the IRS-1 expression was obviously decreased (P< 0.05). After treatment, compared with the model group, there was no significant body weight increase in the Sutherlandia group or the pi-oglitazone group. Indicators of blood glucose, TG, TC in the Sutherlandia group and the pioglitazone group were ob-viously decreased with no statistical difference (P< 0.05 or P< 0.01). In the Sutherlandia group and the pioglitazone group, IRS-1 expression of skeletal muscle in rats was obviously increased with no statistical difference (P< 0.05). It was concluded that type 2 diabetes rats induced by intraperitoneal injection of STZ combined with high fat feed method can induce glucolipid metabolism disorders. Traditional South African herb Sutherlandia can obviously reduce the blood sugar level, improve blood lipid metabolism, and improve the level of insulin. Sutherlandia can improve the IRS-1 expression of skeletal muscle in rats, relief the insulin resistance, and lower blood sugar. However, the effect of its exact ways required further in-depth study.

Adenosquamous carcinoma of the lung is a rare pathological type of non-small cell lung cancer. Adenosquamous carcinoma of the lung has a high degree of malignancy and poor clinical prognosis. Medical treatment plays an important role in patients who have lost the opportunity of operation or have recurrence and metastasis after operation. Platinum based chemotherapy is still the cornerstone of medical treatment. It is recommended that routine driver gene testing should be performed for patients with advanced lung adenosquamous carcinoma. For patients with sensitive driver genes, targeted therapy has a definite effect in some patients. Studies on markers related to immune checkpoint inhibitors suggest that patients with lung adenocarcinoma may benefit from immunotherapy, but there is still a lack of clinical practice data.

Objective:To investigate the clinicopathological characteristics and possible pathogenesis of immune checkpoint inhibitor (ICI) induced myocarditis, and to improve understanding of this new type of myocarditis.Methods:Two cases of ICI induced myocarditis with endomyocardial biopsy available for review were selected from the cases with immune-related adverse events treated by ICI in Peking University First Hospital, Beijing, China from 2020 to 2022. The clinical data, histomorphological characteristics, PD-L1 expression of cardiomyocytes, and classification of inflammatory cells in two cases of ICI-induced myocarditis were analyzed. Relevant literature was reviewed.Results:Case 1 was a 64-year-old male diagnosed with gastric signet ring cell carcinoma. Case 2 was a 56-year-old male ad diagnosed with lung squamous cell carcinoma. Both patients developed acute myocarditis during PD-1 inhibitor treatment, and the disease progressed rapidly. Case 2 was more serious than case 1. Endomyocardial biopsy showed definite cardiomyocytic injury and prominent inflammatory infiltration in both cases, which met the full Dallas criteria for myocarditis. The degenerated and necrotic cardiomyocytes accounted for about 10% of the tissues in case 1 and 30% in case 2, respectively. In case 1, the inflammatory cells counted in the densest area were about 150/HPF, comprised of CD20 + cells (about 5/HPF), CD3 + cells (about 60/HPF), CD8 + cells (about 50/HPF) and CD68 + cells (about 70/HPF). In case 2, the inflammatory cells counted in the densest area were about 350/HPF, comprised of CD20 + cells (0/HPF), CD3 + cells (about 100/HPF), CD8 + cells (about 90/HPF) and CD68 + cells (about 200/HPF). In both cases, PD-L1 + cardiomyocytes aggregated in the inflammatory lesions, and the percentage was about 8% and 30% in case 1 and case 2, respectively. Conclusions:ICI-induced myocarditis is frequently acute onset, severe symptoms, and rapid progression. The histological morphology meets the full Dallas criteria for myocarditis. Expression of PD-L1 in cardiomyocytes can be detected in the inflammatory lesions. The inflammatory cells are comprised of CD8 + T lymphocytes and macrophages and the number of macrophages significantly exceeds that of lymphocytes. Combined with the pathological characteristics and the history of ICI treatment, the diagnosis of ICI-induced myocarditis can be made.

Objective:To study the effect of Compound Duzhong Jiangu Granules on joint function, quality of life and inflammatory factors in patients with Kashin-Beck disease.Methods:By group design, 135 patients with Kashin-Beck disease in Chongxin County, Pingliang City, Gansu Province, were selected and divided into intervention group (100 cases treated with Compound Duzhong Jiangu Granules, 12 g/bag, 1 bag/time, 3 times/day, treatment for 1 month) and control group (35 cases treated with ibuprofen, 0.3 g/capsule, 1 capsule/time, 2 times/day, treatment for 2 weeks) according to the randomized, single-blind principle. The changes of joint dysfunction index score, joint function improvement rate, total scores of Kashin-Beck disease quality of life (KBDQOL) and scores of each dimension were analyzed before treatment, 1 month and 3 months after treatment. The serum levels of interleukin-6 (IL-6), nuclear factor κB (NFκB)-p65, inducible nitric oxide synthase (NOS2), nitric oxide (NO), and prostaglandin E2 (PGE2) were detected by enzyme-linked immunosorbent assay before treatment and 1 month after treatment.Results:The total scores of joint dysfunction index of the intervention group and the control group 1 month after treatment and 3 months after treatment were lower than those before treatment, but 3 months after treatment was higher than 1 month after treatment ( P < 0.001). One month after treatment, the total effective rates of joint function improvement in the intervention group and the control group were 68.00% (68/100) and 54.55% (18/33), respectively; 3 months after treatment, the total effective rates of the intervention group and the control group were 36.00% (36/100) and 39.39%(13/33), respectively. The total scores and scores of each dimension of KBDQOL were not significant for the main effect of the group ( P > 0.05), but significant for the main effect at the time point ( P < 0.05), and there was no interactive effect ( P > 0.05). There were significant differences in the scores of social support and mental status dimensions in the intervention group at different time points ( P < 0.001). And in the intervention group, the total score, the scores of physical function, activity limitation, economy and overall health dimensions were statistically significant between before treatment and 1 month after treatment, 1 month after treatment and 3 months after treatment( P < 0.05); however, there was no significant difference between before treatment and 3 months after treatment ( P > 0.05). One month after treatment, the serum PGE2 levels of both groups were decreased ( P < 0.05), and there was no difference in other inflammatory factors at different groups and time points ( P > 0.05). Conclusion:Compound Duzhong Jiangu Granules can effectively inhibit the inflammatory reaction of patients with Kashin-Beck disease, promote the improvement of joint function and improve the quality of life in various aspects.

Pulmonary mucinous adenocarcinoma is a subtype of lung adenocarcinoma, among which invasive mucinous adenocarcinoma (IMA) is the most common subtype and is easily misdiagnosed as pneumonia. Its etiology and pathogenesis are unclear and may be related to gene mutations and other factors. Due to its relative rarity and few related studies, guidelines do not provide advices on its treatment. KRAS mutations are common in IMA patients, and Sotorasib may be effective against KRAS G12C mutated IMA. NRG1 fusion is considered to be an important driver of IMA, and afatinib may be effective in treating IMA with NRG1 fusion/rearrangement. PD-L1 expression is very low in IMA patients, while B7-H3 expression is high, so B7-H3 may be a potential immunotherapeutic target.