Heterotypic cell-in-cell structures(heCICs)mediate unique non-autonomous cell death,which are widely involved in a variety of important pathological processes,such as tumorigenesis,pro-gression and clinical prognosis.Methylenetetrahydrofolata dehydrogenase 2(MTHFD2),one of the key enzymes of one-carbon metabolism,is highly expressed in a variety of tumor cells.In this study,in order to investigate the effect of MTHFD2 on the formation of heCICs,liver cancer cells and immune cells were first labeled separately by live cell dyes,and the heCIC model was established by using fluorescence mi-croscopy for cell imaging and analysis.After transiently knocking down MTHFD2 in cells by RNAi,we found that the ability of PLC/PRF/5 and Hep3B to form heCICs with immune cells was significantly in-creased(all P＜0.01).MTHFD2 recombinant expression plasmid was constructed by the homologous re-combination method,and MTHFD2 overexpression cell lines were further constructed.Then,the effect of MTHFD2 overexpression on the ability to form heCICs was detected by co-culturing the overexpression cell lines with immune cells.The results showed that the rate of heCIC formation was significantly re-duced after overexpression of MTHFD2(all P＜0.001).In conclusion,this study demonstrated that MTHFD2 is a negative regulator of heCIC formation,providing a research basis for targeting MTHFD2 to promote heCIC formation and enhance the in-cell killing of immune cells.

PURPOSE: Robot-assisted technology is a forefront of surgical innovation that improves the accuracy of total knee arthroplasty (TKA). But whether the accuracy of surgery can improve the clinical efficacy still needs further research. The purpose of this study is to perform three-dimensional (3D) analysis in the early postoperative period of patients who received robot-assisted total knee arthroplasty (RATKA), and to study the trend of changes in gait parameters after RATKA and the correlation with the early clinical efficacy. METHODS: Patients who received RATKA in the Center of Joint Surgery, the First Hospital Affiliated to Army Military Medical University from October 2020 to January 2021 were included. The imaging parameters, i.e., hip-knee-ankle angle, lateral distal femoral angle, medial proximal tibial angle, posterior condylar angle were measured 3 months post-TKA. The 3D gait analysis and clinical efficacy by Western Ontario Mac Master University Index (WOMAC) score were performed pre-TKA, 3 and 6 months post-TKA. The differences in spatiotemporal parameters of gait, kinetic parameters, and kinematic parameters of the operated limb and the contralateral limb were compared. The correlation between gait parameters and WOMAC scores was analyzed. Paired sample t-test and Wilcoxon rank-sum test were used to analyze the difference between groups, and Spearman correlation coefficient was used to analyze the correlation. RESULTS: There were 31 patients included in this study, and the imaging indexes showed that all of them returned to normal post-TKA. The WOMAC score at 3 months post-TKA was significantly lower than that pre-TKA, and there was no significant difference between at 3 and 6 months. The 3D gait analysis results showed that the double support time of the operated limb reduced at 3 and 6 months (all p < 0.05), the maximum extension and maximum external rotation of the knee joint increased at stance phase, and the maximum flexion angle, the range of motion and the maximum external rotation increased at swing phase. Compared with the preoperative data, there were significant improvements (all p < 0.05). Compared with the contralateral knee joint, the maximum external rotation of the knee joint at swing phase was smaller than that of the contralateral side, and the maximum flexion and extension moment was greater than that of the contralateral knee. The maximum external rotation moment of the joint was greater than that of the contralateral knee joint (p < 0.05). There was a negative correlation between the single support time pre-TKA and the WOMAC score at 3 months (p = 0.017), and the single support time at 3 months was negatively correlated with the WOMAC score at 6 months (p = 0.043). The cadence at 6 months was negatively correlated with the WOMAC score at 6 months (p = 0.031). The maximum knee extension at stance phase at 6 months was negatively correlated with the WOMAC score at 6 month (p = 0.048). The maximum external rotation at stance phase at 6 months was negatively correlated with the WOMAC score at 6 months (p = 0.024). CONCLUSION The 3D gait analysis of RATKA patients is more sensitive than WOMAC score in evaluating the clinical efficacy. Trend of changes in gait parameters shows that the knee joint support, flexion and extension function, range of motion, external rotation and varus deformity moment of the patient were significantly improved at 3 months after surgery, and continued to 6 months after surgery. Compared with the contralateral knee, the gait parameters of the operated limb still has significant gaps in functionality, such as the external rotation and flexion and extension. The single support time, cadence, knee extension, and knee external rotation of the operated limb have a greater correlation with the postoperative WOMAC score. Postoperative rehabilitation exercises should be emphasized, which is of great value for improving the early efficacy of RATKA.
Humans ; Arthroplasty, Replacement, Knee ; Gait Analysis ; Robotics ; Osteoarthritis, Knee/surgery* ; Knee Joint/surgery* ; Treatment Outcome ; Range of Motion, Articular ; Biomechanical Phenomena

OBJECTIVES: To explore the application of whole-genome sequencing (WGS) in the rapid clinical diagnosis of critically ill neonates. METHODS: The critically ill neonates who admitted to the neonatal intensive care unit of Children's Hospital of Fudan University and underwent WGS from August to September, 2019 were enrolled in this prospective study. The genetic testing results and clinical outcome were analyzed with reference to the sequencing data and clinical features of the neonates. RESULTS: A total of 15 neonates were tested, among whom there were 9 boys and 6 girls. The main reason for hospitalization included abnormal breathing in 7 neonates, poor response in 2 neonates, feeding difficulty in 2 neonates, fever in 1 neonate, hypothermia in 1 neonate, preterm birth in 1 neonate, and convulsion in 1 neonate. The mean turn-around time was 4.5 days for WGS. Finally a genetic diagnosis was obtained for 3 neonates, with a positive diagnostic rate of 20% (3/15). Among the 3 neonates, 2 neonates were withdrawn from the treatment due to severe conditions and 1 neonate died on the day when the sample was sent for genetic testing, whose etiology could be explained by the results of genetic testing. CONCLUSIONS WGS technique can provide a timely and effective diagnosis for critically ill neonates suspected of genetic diseases and provide genetic evidence for clinical treatment of critically ill cases.
Infant, Newborn ; Male ; Child ; Female ; Humans ; Critical Illness ; Prospective Studies ; Premature Birth ; Dyspnea ; Fever

Objective: To evaluate the safety and efficacy of transcatheter aortic valve implantation (TAVI) with the novel Prizvalve^® system in treating severe aortic stenosis. Methods: This is a single-center, prospective, single-arm, observational study. A total of 11 patients with severe aortic stenosis with high risk or inappropriate for conventional surgical aortic valve replacement (SAVR) were included, and TAVI was achieved with the Prizvalve^® system between March 2021 and May 2021 in West China Hospital. Transthoracic echocardiography (TTE) was performed immediately after prosthesis implantation to evaluate mean transaortic gradient and maximal transaortic velocity. The device success rate was calculated, which was defined as (1) the device being delivered via the access, deployed, implanted and withdrawn, (2) mean transaortic gradient<20 mmHg (1 mmHg=0.133 kPa) or a maximal transaortic velocity<3 m/s post TAVI, and without severe aortic regurgitation or paravalvular leak post TAVI. TTE was performed at 30 days after the surgery, and all-cause mortality as well as the major cardiovascular adverse events (including acute myocardial infarction, disabling hemorrhagic or ischemic stroke) up to 30 days post TAVI were analyzed. Results: The age of 11 included patients were (78.1±6.3) years, with 8 males. A total of 10 patients were with NYHA functional class Ⅲ or Ⅳ. Devices were delivered via the access, deployed, implanted and withdrawn successfully in all patients. Post-implant mean transaortic gradient was (7.55±4.08) mmHg and maximal transaortic velocity was (1.78±0.44) m/s, and both decreased significantly as compared to baseline levels (both P<0.05). No severe aortic regurgitation or paravalvular leak was observed post TAVI. Device success was achieved in all the 11 patients. No patient died or experienced major cardiovascular adverse events up to 30 days post TAVI. Mean transaortic gradient was (9.45±5.07) mmHg and maximal transaortic velocity was (2.05±0.42) m/s at 30 days post TAVI, which were similar as the values measured immediately post TAVI (both P>0.05). Conclusions: TAVI with the Prizvalve^® system is a feasible and relatively safe procedure for patients with severe aortic stenosis and at high risk or inappropriate for SAVR. Further clinical studies could be launched to obtain more clinical experience with Prizvalve^® system.
Aged ; Aged, 80 and over ; Aortic Valve ; Aortic Valve Stenosis/surgery* ; Heart Valve Prosthesis ; Heart Valve Prosthesis Implantation ; Humans ; Male ; Prospective Studies ; Transcatheter Aortic Valve Replacement/methods* ; Treatment Outcome

ObjectiveTo investigate the effect of Draconis Sanguis petroleum ether fraction （DSPEF） on the proliferation， apoptosis， migration， and autophagy of human gastric cancer HGC-27 and MGC-803 cells， and preliminarily elucidate its molecular mechanism. MethodCell counting kit-8 （CCK-8） assay was used to detect the effect of DSPEF at different concentrations （0， 20， 40， 60， 80 mg·L^-1） on the proliferation of HGC-27 and MGC-803 cells after 24， 48， 72 h. Hoechst staining and flow cytometry were used to explore the effects of DSPEF at different concentrations on the apoptosis and apoptosis rate of HGC-27 and MGC-803 cells after 48 h treatment， respectively. The wound healing assay and acridine orange staining were used to investigate the effects of DSPEF on the migration and autophagy of HGC-27 and MGC-803 cells， respectively. Western blot was used to detect the expression levels of signaling pathway-related proteins in HGC-27 and MGC-803 cells treated with DSPEF for 48 h. ResultCompared with the control group， DSPEF（30 mg·L^-1） inhibited the proliferation and migration of HGC-27 and MGC-803 cells in a concentration- and time-dependent manner （P<0.05）， and induced the apoptosis （P<0.01） and autophagy of HGC-27 and MGC-803 cells. DSPEF （60 mg·L^-1） down-regulated the protein levels of phosphorylated mammalian target of rapamycin （p-mTOR） （P<0.05， P<0.01） and down-regulated phospho-signal transducer and activator of transcription 3 （p-STAT3） in HGC-27 and MGC-803 cells （P<0.01）， suggesting that DSPEF presumedly inhibited the proliferation and migration of human gastric cancer HGC-27 and MGC-803 cells and induced their apoptosis and autophagy by inhibiting the mTOR/STAT3 signaling pathway. ConclusionThe down-regulation of the mTOR/STAT3 signaling pathway may be involved in the anti-gastric cancer effect of DSPEF. This study is expected to provide a reference for the investigation of the anti-tumor effect of Draconis Sanguis.

Objective: To explore the protective effects of anthrahydroquinone-2,6-disulfonate (AH 2 QDS) on the kidneys of paraquat (PQ) poisoned rats via the apelin-APJ pathway. Methods: Male Sprague Dawley rats were divided into four experimental groups: control, PQ, PQ+sivelestat, and PQ+AH 2 QDS. The PQ+sivelestat group served as the positive control group. The model of poisoning was established via intragastric treatment with a 20% PQ pesticide solution at 200 mg/kg. Two hours after poisoning, the PQ+sivelestat group was treated with sivelestat, while the PQ+AH 2 QDS group was given AH 2 QDS. Six rats were selected from each group on the first, third, and seventh days after poisoning and dissected after anesthesia. The PQ content of the kidneys was measured using the sodium disulfite method. Hematoxylin-eosin staining of renal tissues was performed to detect pathological changes. Apelin expression in the renal tissues was detected using immunofluorescence. Western blotting was used to detect the expression levels of the following proteins in the kidney tissues: IL-6, TNF-α, apelin-APJ (the apelin-Angiotensin receptor), NF-κB p65, caspase-1, caspase-8, glucose-regulated protein 78 (GRP78), and the C/EBP homologous protein (CHOP). In in vitro study, a PQ toxicity model was established using human tubular epithelial cells treated with standard PQ. Twenty-four hours after poisoning, sivelestat and AH 2 QDS were administered. The levels of oxidative stress in human renal tubular epithelial cells were assessed using a reactive oxygen species fluorescence probe. Results: The PQ content in the kidney tissues of the PQ group was higher than that of the PQ+AH 2 QDS group. Hematoxylin-eosin staining showed extensive hemorrhage and congestion in the renal parenchyma of the PQ group. Vacuolar degeneration of the renal tubule epithelial cells, deposition of crescent-like red staining material in renal follicles, infiltration by a few inflammatory cells, and a small number of cast formation were also observed. However, these pathological changes were less severe in the PQ+sivelestat group and the PQ+AH 2 QDS group (P<0.05). On the third day after poisoning, immunofluorescence assay showed that the level of apelin in the renal tissues was significantly higher in the PQ+AH 2 QDS group than in the PQ group. Western blotting analysis results showed that IL-6, TNF-α, NF-κB p65, caspase-1, caspase-8, GRP78, and CHOP protein levels in the PQ group were higher than in the PQ+AH 2 QDS group (P<0.05). The expression of apelin-APJ proteins in the PQ+AH 2 QDS group was higher than in the PQ+sivelestat and PQ groups (P<0.05); this difference was significant on Day 3 and Day 7. The level of oxidative stress in the renal tubular epithelial cells of the PQ+AH 2 QDS group and the PQ+sivelestat group was significantly lower than in the PQ group (P<0.05). Conclusions: This study confirms that AH 2 QDS has a protective effect on PQ-poisoned kidneys and its positive effect is superior to that of sivelestat. The mechanism of the protective effects of AH 2 QDS may be linked to reduction in cellular oxidative stress, PQ content of renal tissue, inflammatory injury, endoplasmic reticulum stress, and apoptosis. AH 2 QDS may play a role in the treatment of PQ poisoning by upregulating the expression of the apelin-APJ.

Ulcerative colitis (UC) is a chronic non-specific inflammatory bowel disease, listed as a modern refractory disease by the World Health Organization, which is difficult to recover, whereas it is easy to be attacked repeatedly. UC pathogenesis is closely related to gut microbiota dysbiosis. The gut microbiota interacts with bile acids (BAs), short-chain fatty acids (SCFAs), tryptophan, and other metabolism, immune system, intestinal barrier, etc., which regulate each other and affect the occurrence and development of UC. The active ingredients of traditional Chinese medicine (TCM), single herb and its extracts, and formulae can effectively alleviate UC symptoms by regulating the diversity, structure, composition, and metabolites of gut microbiota. In this review, the TCM based on the regulation of gut microbiota in the treatment of UC and its related mechanism for nearly three years was summarized.

Objective:We compare the consistency, similarities and differences of operating procedures, data and conclusions of air-charged catheters(ACC) and water-filled catheters(WFC), as simultaneously using ACC and WFC in pressure-flow study(PFS).Methods:This study was a prospective, synchronously controlled study, including eligible patients who underwent PFS in the Department of Urology, Beijing Chaoyang Hospital from January 2021 to March 2021. Inclusion criteria: ① Patients need PFS for lower urinary tract symptoms like frequency of urination, urgent urination, urinary incontinence and dysuria; ② Age over 18 years old. Exclusion criteria: ① Unable to complete or cooperate during the urodynamic test; ② Patients with severe urethral stricture or acute stage urinary tract infection; ③ Pregnant women. The bladder pressure was measured continuously by using a 7FDR T-DOC ?AC three-chamber bladder pressure tube, which linked to ACC sensor and improved WFC pressure conduction module. At the same time, 7FA T-DOC ?AC single-lumen rectal pressure tube and 7F Labori-CAT411 double-lumen water sac abdominal pressure tube was used to measure the ACC and WFC rectum pressure, respectively. We recorded the Pdet, Pves and Pabd measured by ACC and WFC, at the point of initial sitting position, bladder filling at 100 ml, 150 ml, 200 ml, cough, Q max, maximum Pdet and the end of urination, and compared the mean values, differences, and consistencies of our data. Result:A total of 63 patients (26 female, 37 male) were included in this study, with an average age of 59.19 years (25-86 years old). During bladder filling phase, the mean values of Pves measured by ACC and WFC were 30.78/24.67cmH 2O (initial sitting position), 29.79/25.13cmH 2O (100 ml), 30.87/25.90cmH 2O (150 ml) and 30.95/26.17cmH 2O(200 ml), respectively, the mean value of Pabd were 30.03/24.17cmH 2O (initial sitting position), 28.81/21.78cmH 2O (100ml), 28.89/21.38cmH 2O (150ml), 28.44/21.60cmH 2O (200ml), respectively, and were significantly different at each sampling point ( P<0.01). During urination period, no significant differences were found in data( P>0.05), and the data measured with ACC and WFC system have good consistency. There were significant differences in Pves(mean 57.30/49.95 cmH 2O, respectively) and Pdet(mean 54.21/43.10 cmH 2O, respectively) between ACC and WFC in cough ( P<0.01), but there was a strong linear correlation between these data between two systems(R 2=0.792 in Pves and 0.756 in Pabd). Bland-Altman analysis showed that detrusor pressure at the maximum urine flow rate maintained good consistency between ACC and WFC, which 95% CI was -13.9 cmH 2O to 15.8 cmH 2O. Conclusions:In PFS, although the ACC measurement values (Pves and Pabd) during the filling phase are higher than those WFC readings, but the absolute measurement difference is small, so there is no practical meaning in clinical practice. There was no significant difference in detrusor pressure measured during voiding phase, which indicated that the urodynamic judgment and clinical conclusions of the two systems are highly consistent in judgment of the detrusor contractility and the bladder outlet obstruction.

Objective:To summarize long-term outcome of sacral neuromodulation (SNM) for refractory interstitial cystitis/bladder pain syndrome (IC/BPS).Methods:Between January 2013 and October 2020, 28 patients with IC / BPS who received SNM in Beijing Chaoyang Hospital and Hebei Yanda Hospital were retrospectively analyzed. There were 5 males and 23 females, with median age 63.00 (47.50, 66.75) years old. The urgency score was 4 (3, 4) points, 24-hour micturition frequency was 26 (20.50, 32.50) times, nocturia was 9 (7, 12) times, single urine volume was 59.00 (41.25, 79.50) ml, VAS score was 9.0 (8.0, 9.0) points, O′Leary-Sant score was 31.00 (20.25, 33.00) points, and single maximum urine volume was 100.0 (80.0, 127.5) ml. The improvement of symptoms before operation, test period and last follow-up were compared.Results:The urgency score was 2 (1, 3), the 24-hour micturition frequency was 17.00 (15.00, 22.75), the frequency of nocturia was 5.5 (4.0, 7.0), the single urine volume was 87.50 (70.25, 110.00) ml, the VAS score was 4.0 (3.0, 6.0) and the O′Leary-Sant score was 20.00 (17.00, 23.00) in 28 patients during the test period, which were significantly improved compared with those before operation ( P < 0.05). There was no significant difference in the single maximum urine volume of 135.0 (102.5, 160.0) ml between the two groups ( P > 0.05). 28 patients received SNM Ⅱ permanent stimulator implantation. The median follow-up time was 29.5 (21.25, 61.75) months. Among the 28 patients, 3 patients underwent cystectomy and ileal conduit after removal of the complete SNM system due to the unsatisfied results. Twenty-five cases (89.3%) were still treated with SNM. Among them, 6 cases accepted combinative therapy of oral medicine (antihistamines, sodium pentose polysulfate, hormones, immunosuppressants, etc.). Two cases accepted combinative therapy of intravesical instillation, including heparin in one case and sodium hyaluronate in the other one. Three cases accepted combinative therapy of botulinum toxin injection.One case accepted combinative therapy of bladder augmentation. Two cases accepted combinative therapy of traditional Chinese medicine (TCM). One case accepted combinative therapy of oral medicine and TCM. The remaining 10 cases didn't accept any treatment. Among them, 3 cases were still treated with SNM even though they were not satisfied with the effect, including 1 case due to electrode displacement. At the last follow-up of 25 patients, the urgency score was 2 (1, 3), the 24-hour micturition frequency was 16.50 (13.00, 19.75), the frequency of nocturia was 5.5 (4.0, 9.0), the single urine volume was 105.0 (72.5, 120.0) ml, the VAS score was 4.5 (3.0, 6.0) and the O'Leary Sant score was 16.00 (14.00, 22.50), which were significantly improved compared with those before operation ( P < 0.05), but no difference with those during test period ( P>0.05). There was no significant difference in the single maximum urine volume of 125.0 (102.5, 150.0) ml at the last follow-up compared with preoperative and test period ( P>0.05). Conclusions:As a treatment option for refractory IC / BPS, SNM can improve the symptoms of most patients and maintain good long-term efficacy combined with other.