Objective: To establish a method for the genotyping of fetal M blood group antigen by extracting cell-free fetal DNA (cff-DNA) from maternal plasma, so as to guide the management of M antigen-negative pregnant women with IgG anti-M antibody during pregnancy. Methods: A realtime fluorescent quantitative PCR (realtime PCR) method was established. The specificity and sensitivity of the method were validated by dilution of genomic DNA. Subsequently, a total of 12 M antigen-negative pregnant women were enrolled. The cff-DNA was extracted from maternal plasma, and fetal M antigen genotyping was performed by realtime PCR. Fetuses were classified as M-positive or M-negative according to the presence or absence of amplification curve. The accuracy of the method was validated by comparing fetal M antigen genotyping results with the serological results using the cord or peripheral blood of the neonate at birth. Results: Among the 12 M antigen-negative pregnant women, anti-M was detected in five cases, of which four cases had IgG anti-M, and one case had fetal anemia. The results of fetal M antigen genotyping showed that 9 cases were M-positive (9/12, 75%) and 3 cases were M-negative (3/12, 25%). Serological results of blood samples collected after birth from four M-positive fetuses and one M-negative fetus were consistent with the genotyping results. Conclusion: We have, for the first time, established a non-invasive prenatal genotyping method for fetal M antigen using maternal plasma cff-DNA, and preliminarily demonstrated the feasibility of this method.

Objective:To observe the effects of moxibustion at Shenshu(BL23)and Zusanli(ST36)on Toll-like receptor 4(TLR4)-myeloid differentiation factor 88(Myd88)signaling pathway-mediated inflammatory factors in the synovial tissue of ankle joints of rats with rheumatoid arthritis(RA),and to explore the molecular and biological mechanisms underlying the anti-inflammatory and analgesic effects.Methods:A total of 24 male Sprague-Dawley(SD)rats were randomly divided into a normal group,a model group,and a moxibustion group,with 8 rats in each group.The RA model was established with exposure to wind,cold,and damp environmental factors,along with Freund's complete adjuvant.After three days of modeling,mild moxibustion was applied to bilateral Shenshu(BL23)and Zusanli(ST36)in the moxibustion group using moxa sticks of 0.9 cm in diameter for 30 min each time,once a day for 14 d.Structural changes in the synovial tissue and cells were then observed using hematoxylin-eosin staining and transmission electron microscopy,while immunohistochemistry analysis was used to detect tumor necrosis factor(TNF)-α,interleukin(IL)-17,IL-1β,and IL-6 levels.Moreover,the protein expression levels of Myd88,TLR4,and transient potential receptor vanilloid type 1(TRPV1)in the synovial tissue were detected using Western blotting,while their mRNA expression levels were detected using reverse transcription-polymerase chain reaction.Finally,the levels of IL-1β,IL-2,IL-6,IL-17A,and TNF-α in rat serum were detected using enzyme-linked immunosorbent assay.Results:Compared to the normal group,the model group exhibited notable pathological synovial tissue damage,along with significantly higher IL-1β,IL-6,and TNF-α levels(P<0.01)and a slightly higher IL-17 content(P>0.05).Furthermore,the Myd88,TLR4,and TRPV1 protein and mRNA expression levels and serum IL-1β,IL-2,IL-6,IL-17A,and TNF-α levels were all significantly higher in the model group than in the normal group(P<0.01).Compared to the model group,the moxibustion group exhibited a lower degree of synovial tissue pathological damage,along with significantly lower IL-1β,IL-6,and TNF-α levels(P<0.05 or P<0.01)and a lower IL-17 content without statistical significance(P>0.05).Moreover,the Myd88,TLR4,and TRPV1 protein and mRNA expression levels,and serum IL-1β,IL-2,IL-6,IL-17A,and TNF-α levels were all significantly lower in the moxibustion group than in the model group(P<0.01 or P<0.05).Conclusion:Mild moxibustion at Shenshu(BL23)and Zusanli(ST36)can effectively inhibit TLR4-MyD88 signaling pathway-mediated inflammatory factor expression in the synovial tissue of ankle joints of RA rats.Furthermore,the effect of moxibustion on synovial tissue inflammation in RA rats may be attributed to TRPV1 channel activation.

Objective:To investigate the characteristics of elastic program of excited shear wave of plantar soft tissue of diabetic patients and its correlation with levels of glucose metabolism.Methods:A total of 92 patients with type 2 diabetes who admitted to Cangzhou Hospital of Integrated Traditional Chinese Medicine and Western Medicine during January 2022 and January 2023 were selected as diabetes group,and another 92 patients in the prediabetes stage were included in the prediabetes group.Both groups of patients used excited shear wave with different frequencies(400,450,500,550,and 600 Hz)under elastic technique of shear wave.The velocities of shear waves at the 1st metatarsal head,the 3rd metatarsal head and calcaneus of plantar soft tissue,and the thickness of soft tissue of these patients were assessed.The fasting plasma glucose(FPG),2-hour postprandial glucose(2hPG),and glycosylated hemoglobin(HbA1c)levels of sugar metabolism level of two groups were measured,and the correlations between velocity of shear wave,thickness of soft tissue,and level of glucose metabolism were further analyzed.The occurrence of diabetic foot ulcer in the diabetes group was investigated by using follow-up.The receiver operating characteristic(ROC)curve was adopted to assess the predictive value of the velocity of shear wave,and the thickness of soft tissue for diabetic foot ulcer.Results:The FPG,2hPG and HbA1c of glucose metabolism level in the diabetes group were(6.48±0.48)mmol/L,(9.64±1.31)mmol/L and(6.21±0.15)%,respectively,all of which were higher than those[(8.03±0.62)mmol/L,(12.26±1.25)mmol/L and(7.66±0.42)%]in the prediabetes group,and the differences were statistically significant(t=18.961,18.961,31.185,P<0.05).The thickness level of soft tissue at the first metatarsal head was(10.42±3.11)mm in the diabetes group,which was less than(11.76±3.24)mm in the prediabetes group,with a statistically significant difference(t=2.862,P<0.05).Spearman correlation analysis showed that the velocity of shear wave at 500 Hz frequency had a strongest correlation with FPG and HbA1c(r=0.320,0.256,P<0.05),respectively.In the follow-up for patients of diabetes group,14 cases occurred ulcers and 78 cases did not occur ulcers.The velocities of shear wave of the first metatarsal head of 14 patients with ulcers were higher than those of patients without ulcers at all tested frequencies(400,450,500,550,and 600 Hz).The area under curve(AUC)value of ROC curve,specificity and sensitivity of 500 Hz were respectively 0.775,88.46%,and 78.57%in predicting plantar ulcer of patients with diabetes.Conclusion:The velocity of shear wave of soft tissue at the first metatarsal head is significantly correlated with abnormal glucose metabolism in patients with type 2 diabetes.The velocity of shear wave appeared strongest correlation with FPG and HbA1c at500 Hz frequency,which has higher predictive value for diabetic foot ulcer.

A 75-year-old male patient with non-small cell lung cancer in the lower lobe of the left lung underwent left lobectomy, followed by 4 cycles of adjuvant chemotherapy with paclitaxel and platinum, 4 cycles of chemotherapy with docetaxel and cisplatin, and immunotherapy with sintilimab, 13 cycles of immunotherapy with sintilimab monotherapy, and 5 cycles of endostatin and sintilimab, spanning a total of 5 years. Due to disease progression, he received the monotherapy with anlotinib, 42 days later, the patient developed intestinal perforation, and laparoscopic ileal perforation repair surgery was performed. During the operation, a perforation with a diameter of about 1 cm was observed in the ileum, indicating the outflow of digestive fluid and a large amount of purulent fluid accumulation in the pelvic cavity; no signs of gastrointestinal tumor metastasis were observed. After surgery, the patient developed abdominal infection and peritonitis, and was given symptomatic and supportive treatments such as anti-infection for 7 days. However, the patient developed severe complications such as lung infection, heart failure, sepsis, and respiratory failure, and died.

Purpose To compare the HRCT findings of lung damage in patients with idiopathic inflammatory myopathies(IIMs)with positive and negative anti-Ro-52 antibody.Materials and Methods Fifty patients with IIMs admitted to Beijing Shijitan Hospital,Capital Medical University from January 2015 to July 2023 were retrospectively analyzed.The patients were divided into anti-Ro-52 antibody negative group(18 cases)and positive group(32 cases),the lung CT findings of the two groups were compared,the distribution of lung lesions and related signs were analyzed.Results The incidence of interstitial lung disease in the anti-Ro-52 antibody-negative group was 11.1%(1 case),which was lower than that in the positive group 75.6%(31 cases)(P=0.001).Among the interstitial lung disease-positive patients,the proportion of lesions in the anti-Ro-52 positive group was more symmetrical(28 cases,87.5%,P=0.022)and peripheral distribution(20 cases,62.5%,P=0.039)than that in the anti-Ro-52 negative group.Non-specific interstitial pneumonia-like lesions were the main lesions in the two groups,including 14 cases(43.8%)in the anti-Ro-52 positive group and 7 cases(38.9%)in the negative group.However,9 cases(28.1%)of the anti-Ro-52 positive group had acute interstitial pneumonia-like lesions and 1 case(5.6%)of the negative group had acute interstitial pneumonia-like lesions.Imaging accompanying signs:anti-Ro-52 positive combination and consolidation(14 cases,43.8%,P=0.009),nodular shadow(17 cases,53.1%,P=0.025),and reverse halo sign(12 cases,37.5%,P=0.021)were more than those in the anti-Ro-52 negative group.Conclusion IIMs patients with positive anti-Ro-52 antibody have a higher incidence of interstitial lung disease.HRCT findings are of great significance for diagnosis and treatment of IIMs.

Most cancers are currently incurable, partly due to abnormal post-translational modifications (PTMs). In this study, we initially used multiple myeloma (MM) as a working model and found that SUMOylation activating enzyme subunit 1 (SAE1) promotes the malignancy of MM. Through proteome microarray analysis, SAE1 was identified as a potential target for bioactive colcemid or its derivative colchicine. Elevated levels of SAE1 were associated with poor clinical survival and increased MM proliferation in vitro and in vivo. Additionally, SAE1 directly SUMOylated and upregulated the total protein expression of p27, leading to LLPS-mediated nuclear export of p27. Our study also demonstrated the involvement of SAE1 in other types of cancer cells, and provided the first monomer crystal structure of SAE1 and its key binding model with colchicine. Colchicine also showed promising results in the Patient-Derived Tumor Xenograft (PDX) model. Furthermore, a controlled clinical trial with 56 MM patients demonstrated the clinical efficacy of colchicine. Our findings reveal a novel mechanism by which tumor cells evade p27-induced cellular growth arrest through p27 SUMOylation-mediated nuclear export. SAE1 may serve as a promising therapeutic target, and colchicine may be a potential treatment option for multiple types of cancer in clinical settings.

Objective:To establish a practical patient-based internal quality control method for valproic acid drug concentration monitoring.Methods:Observational Study. A PBRTQC model based on the exponentially weighted moving average (EWMA) method was established using Python. All results of a total of 28, 757 valproic acid concentration data from February 1, 2023 to January 31, 2024 were collected and split into training set and validation set at a ratio of 80% and 20% respectively. The truncation limit (TL) was optimized by using the winsorized mean method and the trimmed mean method. Different weighting coefficients λ were set. Different TL and different λ were combined with the EWMA algorithm into different patient-based real-time quality control (PBRTQC) models. The optimized models were verified by introducing simulated constant errors (CE) and proportional errors (PE) respectively. The false positive alarm rate (FAR) was used to evaluate specificity, and the average number of patients before error detection (ANPed) was used to evaluate sensitivity. According to the daily test volume and quality target requirements, we comprehensively judged whether the performance evaluation indicators of FAR and ANPed meet the laboratory requirements. Bias detection curve was used for determination of the best model.Results:The parameters of the best PBRTQC model for valproic acid drug concentration monitoring are: trimmed mean method with 1.5 standard deviations (i.e., truncating data outside 1.5 standard deviations of the data mean), λ=0.01. The performance verification result shows that ANPed of CE and PE of this model are both less than 100. The comparison between the EQA results and the EWMA results show that the EWMA method results are comparable to the EQA results.Conclusion:A PBRTQC model for the valproic acid drug concentration monitoring project based on the EWMA method has been successfully established. It is comparable with both IQC and EQA results, which means PBRTQC may be used as a supplement to the quality control of daily quality control products.

Purpose To compare the HRCT findings of lung damage in patients with idiopathic inflammatory myopathies(IIMs)with positive and negative anti-Ro-52 antibody.Materials and Methods Fifty patients with IIMs admitted to Beijing Shijitan Hospital,Capital Medical University from January 2015 to July 2023 were retrospectively analyzed.The patients were divided into anti-Ro-52 antibody negative group(18 cases)and positive group(32 cases),the lung CT findings of the two groups were compared,the distribution of lung lesions and related signs were analyzed.Results The incidence of interstitial lung disease in the anti-Ro-52 antibody-negative group was 11.1%(1 case),which was lower than that in the positive group 75.6%(31 cases)(P=0.001).Among the interstitial lung disease-positive patients,the proportion of lesions in the anti-Ro-52 positive group was more symmetrical(28 cases,87.5%,P=0.022)and peripheral distribution(20 cases,62.5%,P=0.039)than that in the anti-Ro-52 negative group.Non-specific interstitial pneumonia-like lesions were the main lesions in the two groups,including 14 cases(43.8%)in the anti-Ro-52 positive group and 7 cases(38.9%)in the negative group.However,9 cases(28.1%)of the anti-Ro-52 positive group had acute interstitial pneumonia-like lesions and 1 case(5.6%)of the negative group had acute interstitial pneumonia-like lesions.Imaging accompanying signs:anti-Ro-52 positive combination and consolidation(14 cases,43.8%,P=0.009),nodular shadow(17 cases,53.1%,P=0.025),and reverse halo sign(12 cases,37.5%,P=0.021)were more than those in the anti-Ro-52 negative group.Conclusion IIMs patients with positive anti-Ro-52 antibody have a higher incidence of interstitial lung disease.HRCT findings are of great significance for diagnosis and treatment of IIMs.

A 75-year-old male patient with non-small cell lung cancer in the lower lobe of the left lung underwent left lobectomy, followed by 4 cycles of adjuvant chemotherapy with paclitaxel and platinum, 4 cycles of chemotherapy with docetaxel and cisplatin, and immunotherapy with sintilimab, 13 cycles of immunotherapy with sintilimab monotherapy, and 5 cycles of endostatin and sintilimab, spanning a total of 5 years. Due to disease progression, he received the monotherapy with anlotinib, 42 days later, the patient developed intestinal perforation, and laparoscopic ileal perforation repair surgery was performed. During the operation, a perforation with a diameter of about 1 cm was observed in the ileum, indicating the outflow of digestive fluid and a large amount of purulent fluid accumulation in the pelvic cavity; no signs of gastrointestinal tumor metastasis were observed. After surgery, the patient developed abdominal infection and peritonitis, and was given symptomatic and supportive treatments such as anti-infection for 7 days. However, the patient developed severe complications such as lung infection, heart failure, sepsis, and respiratory failure, and died.

Objective:To observe the effects of moxibustion at Shenshu(BL23)and Zusanli(ST36)on Toll-like receptor 4(TLR4)-myeloid differentiation factor 88(Myd88)signaling pathway-mediated inflammatory factors in the synovial tissue of ankle joints of rats with rheumatoid arthritis(RA),and to explore the molecular and biological mechanisms underlying the anti-inflammatory and analgesic effects.Methods:A total of 24 male Sprague-Dawley(SD)rats were randomly divided into a normal group,a model group,and a moxibustion group,with 8 rats in each group.The RA model was established with exposure to wind,cold,and damp environmental factors,along with Freund's complete adjuvant.After three days of modeling,mild moxibustion was applied to bilateral Shenshu(BL23)and Zusanli(ST36)in the moxibustion group using moxa sticks of 0.9 cm in diameter for 30 min each time,once a day for 14 d.Structural changes in the synovial tissue and cells were then observed using hematoxylin-eosin staining and transmission electron microscopy,while immunohistochemistry analysis was used to detect tumor necrosis factor(TNF)-α,interleukin(IL)-17,IL-1β,and IL-6 levels.Moreover,the protein expression levels of Myd88,TLR4,and transient potential receptor vanilloid type 1(TRPV1)in the synovial tissue were detected using Western blotting,while their mRNA expression levels were detected using reverse transcription-polymerase chain reaction.Finally,the levels of IL-1β,IL-2,IL-6,IL-17A,and TNF-α in rat serum were detected using enzyme-linked immunosorbent assay.Results:Compared to the normal group,the model group exhibited notable pathological synovial tissue damage,along with significantly higher IL-1β,IL-6,and TNF-α levels(P<0.01)and a slightly higher IL-17 content(P>0.05).Furthermore,the Myd88,TLR4,and TRPV1 protein and mRNA expression levels and serum IL-1β,IL-2,IL-6,IL-17A,and TNF-α levels were all significantly higher in the model group than in the normal group(P<0.01).Compared to the model group,the moxibustion group exhibited a lower degree of synovial tissue pathological damage,along with significantly lower IL-1β,IL-6,and TNF-α levels(P<0.05 or P<0.01)and a lower IL-17 content without statistical significance(P>0.05).Moreover,the Myd88,TLR4,and TRPV1 protein and mRNA expression levels,and serum IL-1β,IL-2,IL-6,IL-17A,and TNF-α levels were all significantly lower in the moxibustion group than in the model group(P<0.01 or P<0.05).Conclusion:Mild moxibustion at Shenshu(BL23)and Zusanli(ST36)can effectively inhibit TLR4-MyD88 signaling pathway-mediated inflammatory factor expression in the synovial tissue of ankle joints of RA rats.Furthermore,the effect of moxibustion on synovial tissue inflammation in RA rats may be attributed to TRPV1 channel activation.