Osteoarthropathy is a progressive disease that causes the degeneration of articular cartilage due to biological and mechanical factors. The destruction of the articular cartilage matrix and the reduction of articular cartilage cells are the core part of its pathological development, and subchondral bone, synovial membrane and joint capsule are also damaged. The Wnt signaling pathway not only plays an essential role in human development and the maintenance of adult tissue homeostasis, but also functions in the joint formation, articular cartilage development, and the occurrence and development of osteoarthropathy. It has become increasingly popular in joint pathology. In this paper, authors attempted to review the research progress of Wnt signaling pathway and its interaction with other signaling pathways in osteoarthropathy.

OBJECTIVE To investigate the feasibility and application of enzyme-linked bridging assay(ELBA)method to the pharmacokinetic evaluation of antisense strand siRNA drug. METHODS Antisense strand RNAs were diluted in LNCap cell lysates from 5 to 50 000 pmol·L-1 to construct the quantification curves. We transfected the intact double-strand siRNA at a final concentration 100 nmol·L-1 targeting Polo-like kinase into the LNCap cells and investigated the specificity of ELBA quantitating the siRNA antisense strand in cell supernatant,cell lysates and RNA-induced silencing complex( RlSC). Quantification curves were constructed and validated in biological matrices such as plasma (5-25 000 pmol·L-1 )and multiple tissues(liver,heart,spleen,and kidneys)(3-6250 pmol·L-1 ). The prostate specific membrane antigen aptamer siRNA delivery system with the intact siRNA concentration of 15 nmol·kg-1 was prepared. The siRNAs were delivered into the LNCap xenogrant tumor model in C57 mice by tail vein injection. The concentration of siRNA antisense strand was determined in plasma and tissues 30 min post administration by ELBA. RESULTS The quantitative range of antisense strand siRNA in cell lysates was 5-50 000 pmol·L-1 ,and ELBA method could quantify the siRNA antisense strand concentration from cell lysates and RlSC in LNCap cells transfected with double-strand siRNA. ln addition,ELBA could specifically reflect the single antisense strand concentration instead of intact siRNA double strands in plasma. The quantification range of siRNA antisense strand using ELBA in plasma was 5-25 000 pmol·L-1 and 3-3125 pmol·L-1 in tissues. About 30 min post administration of PSMA aptamer-siRNA,the antisense strand of siRNA was distributed mainly to the tumor,liver,kidneys,blood and spleen in sequence. The distribution profile might be attributed to the target delivery and siRNA pharma-codynamics. CONCLUSION The ELBA method is successfully applied to the siRNA antisense strand pharmacokinetic evaluation,which provides an alternative for pharmacokinetic studies of siRNA-based drugs.

AIM:To investigate the effect of signal transducer and activator of transcription 1 ( STAT 1 ) on proliferation and interferon-β(IFN-β) sensitivity of human non-small-cell lung cancer H1299 cells.METHODS:STAT1 or EGFP gene was transfected into H1299 cells by the lentiviral vectors system.The cell number was counted under a mi-croscope and cell proliferation was tested by MTT assay.In addition, the cells transfected with STAT1 and EGFP were trea-ted with IFN-βand cell viability was measured by MTT assay.The protein levels of p-STAT1, ICAM-1 and PCNA were de-tected by Western blot.RESULTS: Over-expression of STAT1 inhibited H1299 cell proliferation (P<0.05).H1299 cells transfected with STAT1 gene had a higher sensitivity to IFN-βthan the control cells transfected with EGFP ( P <0.05).Overexpression of STAT1 increased the protein level of p-STAT1, and reduced IACM-1 expression in H1299 cells. Moreover, STAT1 enhanced STAT1 phosphorylation and downregulated the expression of PCNA in H1299 cells treated with IFN-β.CONCLUSION:STAT1 inhibits the proliferation and enhances the IFN-βsensitivity of non-small-cell lung cancer H1299 cells.

Objective:To summarize the clinical manifestations and color Doppler ultrasonography of benign lymphoepithelial disease (BLL) in lacrimal gland, so as to improve the diagnostic accuracy of lacrimal BLL.Methods:Clinical manifestations and color Doppler flow imaging (CDFI) features of lacrimal BLL in 16 patients (2 males and 14 females, with 31 lesions) who visited the Second Hospital of Hebei Medical University from November 2014 to August 2019 were retrospectively analyzed.Results:The performance for lesions in patients with duration less than 3 months was scattered in irregular low echo, lesion blood flow signals within the rich (Adler Ⅱ level), pathologic examination results showed more lymphocytes were seen in the lacrimal matrix, and the myoepithelium of the lacrimal duct grew to form the epithelium-myocutaneous island. The ultrasonic feasures in patients with course of 3-6 months were in multiple categories such as circular low echo, "honeycomb" change, CDFI showed lesions with a lot of blood flow signals (Adler Ⅲ level), the pathological examination results indicated that there were a large number of lymphocytes and epithelial-musculocutaneous islands in the lacrimal matrix, and the lymphocytes were significantly increased compared with patients with the course of disease less than 3 months. The ultrasound results in patients with a course of more than 6 months showed lesions in multiple categories such as circular low echo, and large low echo, greater than 3 mm in diameter, CDFI showed lesions within the same large amounts of blood flow signals (Adler Ⅲ level), the pathological examination results were consistent with the course of 3 to 6 months. One patient had positive tuberculin test and 11 had higher IgG4 than normal.The number of lymphocytes increased with the prolongation of disease course.Conclusions:With the expert knowledge of color Doppler ultrasonographic characteristics of lacrimal gland BLL and with the serum IgG4 level helps to improve the accuracy of ultrasonic diagnosis.

Objective:To explore the incidence and occurrence time of gallstone disease after bariatric surgery.Methods:Retrospectively analyzed the clinical data of 187 patients with morbid obesity who underwent bariatric surgery in the Department of General Surgery, Beijing Friendship Hospital, Capital Medical University from Dec. 2017 to Aug. 2019. All patients did not receive prophylactic ursodeoxycholic acid. All patients were underwent abdominal ultrasound and MRI examination preoperatively, and at least one abdominal ultrasound, MRI examination postoperatively. The incidence and occurrence time of gallstones and biliary sludge in patients with different bariatric surgery were analyzed respectively. Measurement data conforming to the normal distribution were described as mean ± standard deviation ( Mean± SD). Measurement data consistent with skewed distribution were described as median (lower quartile, upper quartile). Counting data were described as a percentage (%). Kruskal-Wallis test was used for comparison among groups, and then Bonferroni correction was used for pairwise comparison. Results:The follow-up time was up to Dec. 31, 2020, with a median follow-up time of 27.0 (22.0, 31.0) months. Thirty-four patients (18.2%, 34/187) developed gallstones after bariatric surgery. Individually, it was 18.0%(30/167) in LSG group, 22.2%(2/9) in LRYGB group, 11.1%(1/9) in LOAGB group and 50.0%(1/2) in LOAGB revisional surgery group. Eighteen patients (9.6%, 18/187) were found biliary sludge formation, among which 8.4% (14/167), 22.2% (2/9), and 22.2% (2/9) underwent LSG, LRYGB, and LOAGB, respectively. The rates of weight loss and BMI loss in patients with postoperative gallstone-formation were 21.4 (18.7, 23.6)% and 21.4 (18.6, 23.5) %, respectively. Three patients (1.6%, 3/187) had newly developed symptomatic gallstones, and all of them underwent LSG. The mean occurrence time for biliary sludge and gallstone was 85.5 (28.8, 98.8) and 103.5 (93.0, 179.3) days, respectively. Statistical difference in occurrence time was only found between postoperative gallstone and biliary sludge formation ( P=0.009). Conclusion:Without drug intervention, the incidence of gallstone after bariatric surgery was about 18.2% (34/187), which requires close clinical attention. Abdominal ultrasonography and the T2WI sequence of upper abdominal MRI can help to diagnose gallstone and monitor its changes.

Objective:To analyze the clinical characteristics of obese adolescents undergoing metabolic and bariatric surgery for the purpose to guide clinical decision-making and practice of metabolic and bariatric surgery in adolescents.Methods:The perioperative clinical profile of 123 adolescent patients under the age of 21 years who underwent metabolic and bariatric surgery in Beijing Friendship Hospital, Capital Medical University from 2018 to 2021 were extracted, and compared their clinical data with the overall data of 6807 patients in the 2021 GC-MBD Annual Report. Analyzed the clinical characteristics of adolescent weight loss metabolic surgery patients from multiple aspects such as preoperative complications, surgical methods, surgical outcomes, and follow-up. Statistical analysis was conducted using SPSS25.0 software.Results:Among adolescent bariatric surgery patients, the proportion of female patients (74.0%) was higher than that of male patients (26.0%). Except for the incidence of polycystic ovary syndrome (31.9%), which was higher than the overall rate, the proportion of other related diseases or symptoms was low and usually mild. The changes of systolic blood pressure ( Z=-5.73, P<0.001), body weight ( Z=-5.69, P<0.001), umbilical abdominal circumference ( Z=-2.40, P=0.017), glycosylated hemoglobin ( Z=-5.23, P<0.001), fasting insulin ( Z=-2.95, P<0.003), fasting C-peptide ( Z=-4.59, P<0.001), triglyceride ( Z=-2.75, P=0.006) at 3 months after operation were statistically significant compared with those before operation. The changes of systolic blood pressure ( Z=-3.42, P=0.001), body weight ( Z=-5.14, P<0.001), umbilical abdominal circumference ( Z=-2.86, P=0.004) and glycosylated hemoglobin ( Z=-2.67, P<0.008), fasting C-peptide ( Z=-2.09, P=0.037), high-density lipoprotein ( Z=-2.08, P=0.038) at 6 months after operation were statistically significant compared with those before operation. Conclusions:The outcomes of bariatric surgery in obese adolescents are similar to those in adults. The indications and timing of bariatric surgery need to be further explored. In view of the high proportion of females, poor self-management ability and low follow-up compliance of adolescent patients, it is urgent to construct a full-cycle care model suitable for adolescent patients with bariatric surgery to improve their self-management ability, follow-up compliance and long-term clinical outcomes.

Objective To explore the public attitude towards kidney xenotransplantation in China by constructing and validating the prediction model based on xenotransplantation questionnaire. Methods A convenient sampling survey was conducted among the public in China with the platform of Wenjuanxing to analyze public acceptance of kidney xenotransplantation and influencing factors. Using random distribution method, all included questionnaires (n=2 280) were divided into the training and validation sets according to a ratio of 7:3. A prediction model was constructed and validated. Results A total of 2 280 questionnaires were included. The public acceptance rate of xenotransplantation was 71.3%. Multivariate analysis showed that gender, marital status, resident area, medical insurance coverage, religious belief, vegetarianism, awareness of kidney xenotransplantation and whether on the waiting list for kidney transplantation were the independent influencing factors for public acceptance of kidney xenotransplantation (all P<0.05). The area under the curve (AUC) of receiver operating characteristic (ROC) of the prediction model in the training set was 0.773, and 0.785 in the validation set. The calibration curves in the training and validation sets indicated that the prediction models yielded good prediction value. Decision curve analysis (DCA) suggested that the prediction efficiency of the model was high. Conclusions In China, public acceptance of kidney xenotransplantation is relatively high, whereas it remains to be significantly enhanced. The prediction model based on questionnaire survey has favorable prediction efficiency, which provides reference for subsequent research.

Objective To investigate the establishment of a six-gene-edited pig-to-non-human primate kidney xenotransplantation model. Methods The kidney of humanized genetically-edited pig (GTKO/β4GalNT2KO/CMAHKO/hCD55/hCD46/hTBM) was transplanted into a cynomolgus monkey. The survival of the recipient and kidney condition after blood perfusion were observed. The parenchymal echo, blood flow changes, and size of the kidney were monitored on a regular basis. Routine blood test, kidney function test and electrolyte assessment were carried out. Dynamic changes of urine, feces and body mass were monitored. At the end of life, the transplant kidney, heart, liver, spleen, lung, and cecum were collected for pathological examination. Results The recipient died at postoperative 7 d. After blood flow was restored, the kidney was properly perfused, the organ was soft and the color was normal. At the end of the recipient's life, a slight amount of purulent secretion was attached to the ventral side of the kidney, with evident congestion and swelling, showing the appearance of "red kidney". Postoperatively, the echo of renal parenchyma was increased, blood flow was decreased, the cortex was gradually thickened, and a slight amount of effusion surrounded the kidney and abdominal cavity over time. In the recipient, the amount of peripheral red blood cells, hemoglobin, albumin, and platelets was progressively decreased, and serum creatinine level was increased to 308 μmol/L at postoperative 7 d, whereas the K⁺ concentration did not significantly change. Light yellow urine was discharged immediately after surgery, diet and drinking water were resumed within postoperative 3 h, and light yellow and normal-shape stool was discharged. The reddish urine was gradually restored to normal color within postoperative 1 d, which were consistent with the results of the routine urine test. A large amount of brown bloody stool was discharged twice in the morning of 2 d after surgery. Omeprazole was given for acid suppression, and the stool returned to normal at postoperative 4 d. The β₂-microglobulin level was increased to 0.75 mg/L at postoperative 7 d. The body mass was increased by 1.7 kg. Autopsy pathological examination showed interstitial edema and bleeding of the transplant kidney, a large amount of infiltration of lymphocytes and macrophages, infiltration of lymphocytes in the arteriole wall and arterial cavity, accompanied by arteritis changes, lymphocyte infiltration in the cecal stroma and congestion in the spleen tissues. No significant abnormal changes were observed in other organs. Conclusions The humanized genetically-edited pig-to-non-human primate kidney xenotransplantation model is successfully established, and postoperative survival of the recipient is 1 week.

【Objective】 To analyze the changes of activation, function and metabolism of apheresis platelets during storage. 【Methods】 Five groups of apheresis platelets(n=10per group), stored up to 5 days at 20~24℃ with agitation, were sampledat day0(4 hours within collection), 1, 2, 3 and 4, respectively to assess the activation indexes as PAC-1 and CD62P and platelet function was evaluated by thromboelastogram.The platelet counts and related parameters were determined, and the biochemical indexes such as pH, glucose, lactic acid and lactate dehydrogenase reflecting metabolism were measured and statistically analyzed. 【Results】 The CD62P (%) and PAC-1 (%) of apheresis platelets with different storage duration showed significant differences(P<0.05), and CD62P (%) increased significantly on 4thday of storage (P<0.05). There were significant differences in TEG parameters as R(min), K(min), ANG (min)and MA (mm), and MA value increased significantly after1-daystorage (P<0.05). There was no significant difference in Plt and related indexes among the groups (P>0.05). Significant differences were notable in platelet biochemical indexesby groups using variance analysis(P<0.05). The pH value and glucose were the lowest in platelets with 4-day storage, while lactic acid and lactate dehydrogenase were the highest. 【Conclusion】 The storagelesion of apheresis platelets occurs as the activation, function and metabolism of platelets developed as the storage prolonged.

Objective:To screen T-cell exhaustion-related signature genes as the prognostic marker for osteosarcoma and establish a prognostic model for osteosarcoma patients based on weighted gene co-expression network analysis(WGCNA)and Least absolute shrinkage and selection operator(LASSO)-COX regression analysis. Methods:GSE21257 dataset was downloaded from Gene Expression Omnibus(GEO)database for the establishment of the prognostic model for osteosarcoma.4 T-cell exhaustion-related gene sets were downloaded from The Molecular Signatures Database(MisgDB)and their enrichment scores in GSE21257 samples were calculated by single sample gene set enrichment analysis(ssGSEA).WGCNA was carried out to screen the gene module that is highly associated with T-cell exhaustion based on ssGSEA results followed by GO(Gene Ontology)and KEGG(Kyoto Encyclopedia of Genes and Genomes)analysis of the biological processes and signaling transduction pathways that those genes are involved in.The signature genes that are highly associated with the prognosis of osteosarcoma patients were obtained through LASSO-COX regression and a prognostic model was established based on these signature genes.Osteosarcoma-related expression profile data from the GSE21257 and TAEGET datasets on XENA were downloaded from the Gene Expression Omnibus.Clinical information for the training and validation sets was obtained.T-cell exhaustion-related genes were screened using a weighted correlation network analysis.Realtime fluorescence quantitative PCR,COX regression analysis,external dataset and nomogram were used to evaluate the reliability and accuracy of the prognostic model.A immunotherapy-related dataset was used to assess the efficacy of this prognostic model for the prediction of patients'responses to immunotherapy. Results:Analysis results based on the ssGSEA scores showed that T-cell exhaustion-related genes were related to the metastasis and age of osteosarcoma patients.Many T-cell exhaustion-related genes were found to be differentially expressed in metastatic and non-metastatic osteosarcoma patients.1 256 T-cell exhaustion-related genes were identified through WGCNA and these candidate markers were mainly distributed in structures like secretory granule membranes and endocytic vesicles and were involved in T-cell activation.COX regression analysis screened 68 significant prognostic markers out of the 1 256 genes,and 12 signature genes were further confirmed with LASSO-COX regression analysis.A prognostic model was established based on the 12 signature genes.Results of real-time fluorescence quantitative PCR showed a similar trend in the expression of most of the signature genes in different osteosarcoma cell lines.COX regression analysis of the internal and external datasets verified that the risk score calculated with the prognostic model was an independent prognostic factor for osteosarcoma patients,and high-risk score was associated with poor prognosis of the patients.Receiver operating characteristic(ROC)curves demonstrated excellent prognostic efficacy of the model.Nomogram analysis verified the prognostic model is highly accurate and reliable in predicting the prognosis of osteosarcoma patients.Analysis using the immunotherapy-related dataset indicated that this prognostic model could also be used to predict patients'responses to immunotherapy. Conclusion:The 12 signature gene(CD300LB,TRO,SNX3,VENTX,PPM1M,DOT1L,CDC37,NAT9,TRMT1,PPP1R3C,CHTF18 and NSUN5)-based prognostic model can effectively predict the prognosis and responses to immune check-point inhibitors for osteosarcoma patients,which may provide evidence for the prediction of prognosis as well as the selection of immunotherapy plans in clinical practice.