BACKGROUND:Human periodontal ligament stem cells are potential functional cells for periodontal tissue engineering.However,long-term in vitro culture may lead to reduced stemness and replicative senescence of periodontal ligament stem cells,which may impair the therapeutic effect of human periodontal ligament stem cells. OBJECTIVE:To investigate the effect of oxymatrine on the stemness maintenance and osteogenic differentiation of periodontal ligament stem cells in vitro,and to explore the potential mechanism. METHODS:Periodontal ligament stem cells were isolated from human periodontal ligament tissues by tissue explant enzyme digestion and cultured.The surface markers of mesenchymal cells were identified by flow cytometry.Periodontal ligament stem cells were incubated with 0,2.5,5,and 10 μg/mL oxymatrine.The effect of oxymatrine on the proliferation activity of periodontal ligament stem cells was detected by CCK8 assay.The appropriate drug concentration for subsequent experiments was screened.Western blot assay was used to detect the expression of stem cell non-specific proteins SOX2 and OCT4 in periodontal ligament stem cells.qRT-PCR and western blot assay were used to detect the expression levels of related osteogenic genes and proteins in periodontal ligament stem cells. RESULTS AND CONCLUSION:(1)The results of CCK8 assay showed that 2.5 μg/mL oxymatrine significantly enhanced the proliferative activity of periodontal stem cells,and the subsequent experiment selected 2.5 μg/mL oxymatrine to intervene.(2)Compared with the blank control group,the protein expression level of SOX2,a stem marker of periodontal ligament stem cells in the oxymatrine group did not change significantly(P>0.05),and the expression of OCT4 was significantly up-regulated(P<0.05).(3)Compared with the osteogenic induction group,the osteogenic genes ALP,RUNX2 mRNA expression and their osteogenic associated protein ALP protein expression of periodontal ligament stem cells were significantly down-regulated in the oxymatrine+osteogenic induction group(P<0.05).(4)The oxymatrine up-regulated the expression of stemness markers of periodontal ligament stem cells and inhibited the bone differentiation of periodontal ligament stem cells,and the results of high-throughput sequencing showed that it may be associated with WNT2,WNT16,COMP,and BMP6.

ObjectiveTo investigate the effect and mechanism of Taohong Siwutang （TSD） on myocardial ischemia reperfusion injury （MIRI） in ovariectomized （OVX） female mice. MethodAfter the OVX model of female mice was established， the estradiol （E₂） level was detected by enzyme-linked immunosorbent assay （ELISA） to verify the model. Sixty OVX mice were randomly divided into six groups： Sham operation group， model group， low， medium， and high dose groups （9， 18， 36 g·kg^-1） of TSD， and nuclear factor E₂-related factor 2 （Nrf2） inhibitor group， with 10 mice in each group. The MIRI model was verified by a laser speckle flowmeter. The pathological changes in myocardial tissue were detected by 2，3， 5-triphenyltetrazolium chloride （TTC） and hematoxylin-eosin （HE） staining. Serum creatine kinase isoenzyme （CK-MB）， cardiac troponin Ⅰ （CTnⅠ）， malondialdehyde （MDA）， superoxide dismutase （SOD）， interleukin-6 （IL-6）， and interleukin-10 （IL-10） levels were detected by ELISA. The expression of Nrf2 and heme oxygenase-1 （HO-1） were observed by immunofluorescence staining. The expressions of Nrf2， HO-1， apoptotic B-cell lymphomato-2 （Bcl-2）， Bcl-2 associated X protein （Bax）， inflammatory cytokines interleukin-18 （IL-18）， and interleukin-1β （IL-1β） were detected by Western blot. ResultCompared with the sham operation group， the serum levels of CK-MB， CTnⅠ， MDA， and IL-6 in the model group were increased （P<0.01）， and the levels of SOD and IL-10 were decreased （P<0.01）. The damage scores of TTC and HE staining in myocardial tissue were increased （P<0.01）. The expressions of Nrf2 and HO-1 in myocardial tissue by immunofluorescence were decreased （P<0.01）， and the protein expressions of Nrf2， HO-1， and Bcl-2 in myocardial tissue were decreased. The protein expressions of Bax， IL-18， and IL-1β were increased （P<0.01）. Compared with the model group， serum levels of CK-MB， CTnⅠ， MDA， and IL-6 of TSD groups were significantly decreased （P<0.05， P<0.01）， and SOD and IL-10 were significantly increased （P<0.05， P<0.01）. TTC staining and HE staining damage scores of myocardial tissue were significantly decreased （P<0.01）. The expressions of Nrf2 and HO-1 in myocardial tissue by immunofluorescence were increased （P<0.01）. The protein expressions of Nrf2， HO-1， and Bcl-2 were significantly increased （P<0.05， P<0.01）， and those of Bax， IL-18， and IL-1β were significantly decreased （P<0.05， P<0.01）. The effect of the high dose group of TSD was the most significant. The serum levels of CK-MB， CTnⅠ， MDA， and IL-6 in the Nrf2 inhibitor group were significantly increased （P<0.05， P<0.01）， and the levels of SOD and IL-10 were significantly decreased （P<0.05， P<0.01）. The damage scores of TTC and HE staining in myocardial tissue were significantly increased （P<0.01）. The expressions of Nrf2 and HO-1 in myocardial tissue by immunofluorescence were significantly decreased （P<0.01）. The protein expressions of Nrf2， HO-1， and Bcl-2 in myocardial tissue were significantly decreased， and those of Bax， IL-18， and IL-1β were significantly increased （P<0.01）. ConclusionTSD can alleviate MIRI in OVX mice， reduce oxidative stress response and the release of inflammatory factors， and inhibit apoptosis， playing a protective role in OVX mice with MIRI， which may be related to the activation of Nrf2/HO-1 signaling pathway.

Objective:To explore the role of mechanical hemodynamic support (MHS) in mapping and catheter ablation of patients with hemodynamically unstable ventricular tachycardia (VT), report single-center experience in a cohort of consecutive patients receiving VT ablation during MHS therapy, and provide evidence-based medical evidence for clinical practice.Methods:This was a retrospective cohort study. Patients with hemodynamically unstable VT who underwent catheter ablation with MHS at Beijing Anzhen Hospital, Capital Medical University between August 2021 and December 2023 were included. Patients were divided into rescue group and preventive group according to the purpose of treatment. Their demographic data, periprocedural details, and clinical outcomes were collected and analyzed.Results:A total of 15 patients with hemodynamically unstable VT were included (8 patients in the rescue group and 7 patients in the preventive group). The acute procedure was successful in all patients. One patient in the rescue group had surgical left ventricular assist device (LVAD) implantation, remaining 14 patients received extracorporeal membrane oxygenation (ECMO) for circulation support. ECMO decannulation was performed in 12 patients due to clinical and hemodynamic stability, of which 6 patients were decannulation immediately after surgery and the remaining patients were decannulation at 2.0 (2.5) d after surgery. Two patients in the rescue group died during the index admission due to refractory heart failure and cerebral hemorrhage. During a median follow-up of 30 d (1 d to 12 months), one patient with LVAD had one episode of ventricular fibrillation at 6 months after discharge, and no further episodes of ventricular fibrillation and/or VT occurred after treatment with antiarrhythmic drugs. No malignant ventricular arrhythmia occurred in the remaining 12 patients who were followed up.Conclusions:MHS contributes to the successful completion of mapping and catheter ablation in patients with hemodynamically unstable VT, providing desirable hemodynamic status for emergency and elective conditions.

Objective : To explore whether the long⁃chain noncoding RNA associated with genomic instability in colon cancer can predict clinical prognosis and therapeutic. Methods : The R package " limma" was used for differential analysis , and the prognostic risk model was constructed by univariate Cox analysis and multivariate Cox proportional risk regression analysis. The difference in prognosis was evaluated by Kaplan⁃Meier method , and the difference was significant by log⁃rank test. The efficiency of the prognostic model was evaluated using a time⁃dependent area under the subject operating characteristic curve (AUC) . The R package“ pRRophetic ”was used to predict the sensitivity of patients to anticancer drugs. R software package rms was used to build a line graph , and the consistency index of the line graph was calculated. Real⁃time quantitative PCR was used to detect the difference in the expression levels of prognostic protective factors. Results : A total of 22 LncRNAs associated with genomic instabili⁃y in patients with prognosis were obtained , 2 were protective factors for prognosis in patients with colon cancer, and 20 were risk factors for prognosis. A prognosis model composed of LncRNAs associated with genomic instability was constructed , and patients with high risk scores had lower AUCs and shorter median survival. The five⁃year survival AUC predicted by the model was 0. 823 in the training set , 0. 722 in the validation set , and 0. 759 in the overall TCGA colon cancer patient population. Patients with low risk scores had lower half inhibitory concentration (IC50 ) of cisplatin and higher sensitivity (P < 0. 000 1) . The expression of prognostic protective factors in colon cancer tissues was significantly lower than that in adjacent colon cancer tissues. Conclusion A prognostic risk model composed of 8 LncRNAs associated with genomic instability was constructed and verified. In addition , the model can also predict cisplatin drug sensitivity. A histogram was constructed combining the tumor stage and the prognosis model. The predictive ability of this graph for five⁃year survival of colon cancer patients is better than that of traditional histopathological features and prognostic models constructed by predecessors.

Objective: To investigate the effects of retinoid-related orphan receptor γ (RORγ) gene on proliferation and metastasis of human colon cancer cells． Methods: RORγ knockdown cell lines were constructed and the knockdown efficiency was detected by RT-qPCR and Western blot assays ; MTT，colony formation，Transwell and wound healing assays were used to detect cell proliferation and metastasis ; the expression of epithelial-mesenchymal transition (EMT) related proteins was detected by Western blot．The relationship between RORγ gene expression and immune cell infiltration in tumor microenvironment was analyzed using TIMER 2. 0 database. Results : The knockdown of RORγ enhanced the viability (F = 157. 40，P＜0. 01) ，clonogenesis (F = 61. 35，P＜0. 01) ，migration (F = 13. 00，P＜0. 01) ，invasion (F = 21. 26，P＜0. 01) and wound healing ability (F = 877. 2，P＜0. 01) of colon cancer cells，inhibited the expression of E-Cadherin，and promoted the expression of vimentin and N-Cadherin．TIMER 2. 0 database analysis showed that RORγ expression in colon adenocarcinoma ( COAD) tissues was associated with multiple immune cell infiltrates． Conclusion Downregulation of RORγ expression promoted the proliferation and metastasis of colon cancer cells.

Objective:To investigate the predictive value of serum albumin (Alb) levels in the early postoperative period (within 1 hour) for urosepsis after minimally invasive percutaneous nephrolithotomy (mPCNL).Methods:The clinical data of 160 patients treated by single channel holmium laser mPCNL in urolithiasis treatment center of the second hospital of Tianjin Medical University from January 2019 to January 2020 were retrospectively analyzed. The patients were divided into the sepsis group and the non-sepsis group according to whether the patients developed urosepsis after the operation. There were 110 cases of male, and 50 cases of female, with average age of(51.8±11.9), including 68 cases with hypertension, 26 cases with diabetes, 12 patients with history of preoperative fever, 16 cases with history of endoscopic lithotomy surgery, 6 patients with preoperative catheter, 24 patients with positive preoperative urine culture, 12 patients with positive preoperative nitrite volume, 32 cases with preoperative routine urine leucocyte(+ + + ). The operative time and infusion volume were recorded during the operation. Blood samples were collected from all patients within 1 hour after surgery to detect blood routine and serum albumin (Alb), and the change of haemorrhagic white blood cells and serum albumin were calculated. Univariate and multivariate analyses were used to statistically analyze the relevant clinical parameters to identify the independent risk factors for urosepsis after mPCNL surgery. The ROC curve was established and the area under the comparison curve was calculated to analyze the predictive value of independent risk factors for postoperative urosepsis.Results:There were 13 patients (8.1%) in the sepsis group, including 7 males and 6 females, with an average age of (53.1±8.2) years. There were 147 patients (91.9%) in the non-sepsis group, including 103 males and 44 females, with an average age of (51.7±12.1)years. Single factor analysis showed the infectious stones (38.5% and 8.8%), surgery duration [(94.6±26.2)min and(63.6±24.5)min], a positive urine culture (46.2% and 12.2%), positive urinary nitrite (30.8% and 5.4%), strong positive urinary leukocyte (61.5% and 11.3%), postoperative albumin levels [(34.2±2.7)g/L and(40.7±6.2)g/L]and changes of serum albumin levels within one hour after surgery [(18.3±4.6)% and(3.8±14.3)%] between the sepsis group and the non-sepsis group had significant differences ( P<0.05). Multiariable logistic regression analysis showed the strong positive urine leucocyte ( OR=57.704, 95% CI 2.407-1383.427, P=0.012), postoperative blood leucocyte level within one hour after surgery ( OR=2.406, 95% CI 1.154-3.627, P=0.014) and the change of serum albumin levels after surgery ( OR=1.373, 95% CI 1.083-1.740, P=0.009) were independent risk factors for sepsis. ROC curve analysis indicated that the AUC of the change amplitude of serum albumin level was 0.926, which was significantly higher than the AUC of the qualitative results of blood leukocyte and the AUC of urinary leukocyte( P<0.01). Conclusions:The positive urinary leucocyte, the change of serum albumin levels after surgery within one hour and the leucocyte level are independent risk factors for postoperative urosepsis after mPCNL. The change of serum albumin level in the early postoperative period has a strong predictive value for the early diagnosis of postoperative sepsis.

Objective: To investigate the clinical and molecular genetic features of neonatal congenital lipoid adrenal hyperplasia (CLAH) caused by mutations in steroidogenic acute regulatory protein (StAR) encoding gene. Methods: This study retrospectively analyzed the clinical data of a CLAH neonate admitted to Fujian Provincial Maternity and Children's Hospital, Affiliated Hospital of Fujian Medical University in April 2017. StAR gene was analyzed using high-throughput sequencing and Sanger sequencing. Relevant literature retrieved from databases including China National Knowledge Infrastructure (CNKI), Wanfang and PubMed were reviewed, and the reported cases with relatively complete clinical data and results of serum hormone test and StAR gene mutation analysis were collected. Results: The index patient presented with hyperpigmentation and growth retardation soon after birth. Laboratory tests revealed hyponatremia, hyperkalemia, increased serum adrenocorticotrophic hormone (263.4 pmol/L) and decreased 17-hydroxyprogesterone (0.16 ng/ml), dehydroepiandrosterone (<0.95 μmol/L), androstenedione (<1.0 nmol/L), testosterone (<0.025 ng/ml), progesterone (0.02 ng/ml) and cortisol (1.6 μg/ml). High-throughput sequencing showed that the patient carried a compound heterozygous mutation of p.Thr240fs in exon 6 and p.Gln258X in exon 7, inherited from the father and mother, respectively. Sanger sequencing confirmed the diagnosis of CLAH caused by StAR gene mutation. After steroid replacement therapy, the patient's symptoms resolved and the concentrations of electrolytes returned to normal. The neonate was followed up to two years of age and no abnormality was found in physical or neurological development. Two Chinese and 11 English publications were retrieved and altogether 96 cases of neonatal CLAH, including the index one, were reviewed and 42 of them had detailed clinical data. The most common clinical manifestations were skin pigmentation (85.7%, 36/42). Other manifestations included vomiting (35.7%, 15/42) and growth retardation (14.3%, 6/42). All patients with physical examination records had female external genitalia (100.0%, 35/35). The common laboratory abnormalities included hyponatremia (95.2%, 40/42), hyperkalemia (88.1%, 37/42), elevated serum adrenocorticotrophic hormone (100.0%, 37/37) and decreased 17-hydroxyprogesterone (90.5%, 19/21), cortisol (86.2%, 25/29), testosterone (9/10) and dehydroepiandrosterone (14/14). p.Gln258X was the most common StAR gene mutation in neonates in Eastern Asia, including China. Most cases had a good prognosis after appropriate steroid replacement. Conclusions CLAH should be considered for neonates with adrenocortical hypofunction, especially with female phenotypes and low 17-hydroxyprogesterone. Karyotyping and StAR gene analysis may be helpful in diagnosis. Timely and appropriate treatment could improve the prognosis.

Objective:To compare the effectivity and safety of ureteral stenting and percutaneous nephrostomy for patients with upper urinary calculi and sepsis.Methods:From June 2013 to June 2019, 429 patients with upper urinary calculi and urosepsis were accepted in the second hospital of Tianjin Medical University. According to surgical decompression, patients were divided into two groups, ureteral stenting (US, n=304) and percutaneous nephrostomy (PCN, n=125).121 patients were accompanied with septic shock in US group, 56 in PCN group. The effectivity of decompression was analyzed separately in patients with or without shock. For decompression, data included the success rate of decompression, the time of infection related parameters (temperature, blood WBC and CRP) returning to normal and the complications (progress of infection within 30min after decompression, perforation of ureter or pelvis). When urosepsis was cured, ureteroscopic lithotripsy was followed for all patients. The operation time, the stone free rate, the rate of using RIRS and the complications were compared.Results:For patients without septic shock, the success rate of decompression in PCN was higher (68/69)than that of US(165/183)( P =0.025); there was no significant difference in hospital stay for infection control and the time of infection related parameters (temperature, blood WBC and CRP) returning to normal ( P>0.05). The rate of infection progress within 30min in US(25/183) was higher than PCN(3/69)( P=0.036). When ureteroscopic lithotripsy was mentioned, the operation time in US(38.5±6.8 min) was longer than PCN(32.8±4.5 min)( P=0.000), the stone free rate and the rate of using RIRS were lower in PCN( P=0.044, P=0.0002). For patients with septic shock, the success rate of decompression in PCN was higher (55/56)than that of US(106/121)( P=0.022). The rate of infection progress within 30min after decompression was still higher in US ( P=0.048), the time of infection related parameters (temperature, blood WBC and CRP) returning to normal was shorter in PCN ( P=0.000, P=0.003, P=0.000). For lithotripsy, the operation time was longer in US ( P=0.017), the stone free rate and the rate of using RIRS were lower in PCN ( P=0.024, P=0.005). Conclusions:For patients with upper urinary calculi and urosepsis, both ureteral stenting and percutaneous nephrostomy can drainage the pelvis effectively. PCN provides quick recovery, especially when septic shock is involved. For the following ureteroscopic lithotripsy, PCN contributes to less operation time and higher stone free rate, reduces the use of flexible ureteroscope.

Objective:To investigate the clinical characteristics, pathogen distribution and risk factors of late-onset sepsis (LOS) in very low birth weight infants (VLBWI).Methods:In this retrospective case-control study, 107 VLBWIs diagnosed with LOS and hospitalized in Fujian Provincial Maternity and Children's Hospital from January 1, 2010 to December 31, 2015 were enrolled as LOS group. Another 107 VLBWIs without infection were assigned as control group with an allocation ratio of 1 to 1. The clinical data between groups were compared using two-independent sample t-test, Chi-square test, sum-rank test and univariate analysis of variance, and multivariate logistic regression was used to analyze the risk factors of LOS. Results:The incidence of LOS in VLBWI was 8.6% (107/1 239). Among the 107 cases with LOS, 87 recovered with a cure rate of 81.3%. Various clinical presentations were observed, and the most common included lethargy (83/107, 77.6%), abdominal distention (77/107, 72.0%) and dyspnea (76/107, 71.0%). Increased C-reactive protein (CRP) level was the most common laboratory markers (82/107, 76.6%). The blood cultures were positive in 45 (42.1%) cases and the dominant pathogen was Gram-negative bacteria (32/45, 71.1%), especially Klebsiella. The logistic regression analysis showed that mechanical ventilation ( OR=21.181, 95% CI: 1.542-290.948, P=0.022), feeding intolerance ( OR=12.480, 95% CI: 2.602-59.856, P=0.002), combined application of antibiotics before LOS occurs ( OR=22.457, 95% CI: 3.933-128.237, P<0.001), duration of antibiotic treatment before LOS occurs ( OR=1.388, 95% CI: 1.158-1.663, P<0.001) were the independent risk factors of LOS for VLBWI. Conclusions:The clinical presentation of LOS in VLBWI are diverse and non-specific. Increased CRP level is a sensitive laboratory marker. The main pathogen is Gram-negative bacteria. LOS are more prone to occur in VLBWI with mechanical ventilation, feeding intolerance, combined application of antibiotics or long duration of antibiotic treatment.

Objective To summarize the clinical features of neonatal lupus erythematosus (NLE) and to have a better clinical understanding. Methods We retrospectively analyzed the clinical data of 13 patients with NLE who were hospitalized in Fujian Provincial Maternity and Children's Hospital from September 2010 to September 2017. The pathogenesis, clinical manifestations, laboratory examinition, management and long term outcomes of these babies were summarized, and the relevant literatures were also reviewed. Descriptive statistical analysis was performed. ResuLts The 13 NLE cases included eight boys and five girls. Among them, skin lesions, cardiac impairment, hematological problems, hepatobiliary system damage, central nervous system involvement and renal function damage occurred in eight, seven (six cases were atrioventricular block), seven, three, one and one case, respectively. Antinuclear antibodies and anti-Sjogren's syndrome antigen A antibodies were positive in all neonates, anti-Sjogren's syndrome antigen B antibodies were positive in 11, and anti-double-stranded DNA antibodies were positive in two cases. Among the 13 mothers, three were diagnosed with Sjogren's syndrome and two had systemic lupus erythematosus(SLE) before pregnancy, two were diagnosed with Sjogren's syndrome and one developed SLE during pregnancy. Eight babies with skin lesions were asked to avoid light and the skin rash all gradually receded within 1-6 months after birth. Five cases with thrombocytopenia were treated with intravenous immunoglobulin and one anemic baby received erythrocyte transfusion. Within 2 to 3 months, the impaired blood system in these babies were back to normal. For the three babies with abnormal liver functions, hepatic protectants and jaundice relieving agents were given, and 2 to 6 months later they recovered. The 13 patients were followed up for five months to seven years, among which, seven improved with normal growth and development; five still had grade Ⅲ atrioventricular block; one installed an atrial pacemaker at 11 months. Three mothers who were asymptomatic during pregnancy were found to have autoimmune diseases after their babies were diagnosed with NLE, including one case of Sjogren's syndrome and two of SLE. ConcLusions NLE is mainly characterized by skin lesions and congenital heart block, while liver, blood system, central nervous system and other organs may also be involved. For high-risk mothers and babies, timely autoantibody screening and relevant examinations are suggested for early diagnosis and interventions. In addition, long term follow-up is required for affected cases.