OBJECTIVES: To explore the expression profile of circular RNAs (circRNAs) and their potential roles in prognosis and progression of Wilms' tumor (WT). METHODS: Four pairs of WT and adjacent tissues were collected for high-throughput circRNA sequencing to identify the differentially expressed circular RNAs. RT-qPCR was used to verify the expression levels of the top 6 candidate circRNAs in the clinical samples. hsa_circ_0001900 was selected for analysis of its correlation with clinicopathological features and prognosis in 34 patients with WT. Sanger sequencing and RNase R digestion experiments were used to verify the cycling site and structural stability of hsa_circ_0001900 molecule. RESULTS: A total of 23 978 circular RNA molecules were identified in WT tissues by high-throughput circular RNA sequencing, and among them 614 were differentially expressed in WT. hsa_circ_0001900 showed the highest expression level among the differentially expressed circRNAs, which was consistent with the findings in clinical tumor samples and the sequencing results. Correlation analysis showed that hsa_circ_0001900 expression level was positively correlated with WT volume, and the children with high hsa_circ_0001900 expression had a lowered recurrence-free survival rate. The results of Sanger sequencing verified the circular splice site sequence of the molecule, and Rnase R digestion assay confirmed its stable covalent structure. CONCLUSIONS This study presents a comprehensive expression profile of circular RNAs in WT, and the expression level of hsa_circ_0001900 is related to the size of WT and the patients' prognosis, suggesting its possible role as a key driving gene in WT progression.
Humans ; RNA, Circular ; Wilms Tumor/pathology* ; Prognosis ; High-Throughput Nucleotide Sequencing ; Kidney Neoplasms/genetics* ; Sequence Analysis, RNA ; Male ; Female

O-acetyl-L-homoserine (OAH) is a promising platform compound for the production of L-methionine and other valuable compounds, while its low yield and low conversion rate limit the industrial application. To solve these problems, we constructed a strain for high OAH production with the previously constructed L-homoserine producer Escherichia coli HS33 as the chassis by systematic metabolic engineering. Firstly, PEP accumulation, pyruvate utilization, and OAH synthesis pathway (overexpressing aspB, aspA, and thrA^C1034T) were enhanced to obtain an initial strain accumulating 13.37 g/L OAH. Subsequently, the co-factor synthesis genes were integrated to supply reducing power and energy, which increased the yield to 15.79 g/L. The OAH yield of the engineered strain OAH28 was further increased to 17.49 g/L by strengthening the acetic acid reuse pathway, improving the supply of acetyl-CoA, and regulating the expression of MetX from different sources. Finally, in a 5 L fermenter, OAH28 achieved an OAH titer of 47.12 g/L, with a glucose conversion rate of 32% and productivity of 0.59 g/(L·h). The results lay a foundation for increasing the OAH production by metabolic engineering and give insights into the industrial production of OAH.
Metabolic Engineering/methods* ; Escherichia coli/genetics* ; Homoserine/biosynthesis* ; Fermentation

ObjectiveTo evaluate the clinical efficacy of Huayu Tongluo Formula （化瘀通络方， HTF） in patients with mid-stage diabetic kidney disease of blood stasis syndrome and explore its potential mechanisms. MethodsA multi-center， randomized， double-blind， placebo-controlled clinical trial was conducted. Ninety patients of mid-stage diabetic kidney disease of blood stasis syndrome were divided into a control group of 46 cases and a treatment group of 44 cases. Both groups received conventional western medicine treatment， the treatment group additionally taking HTF， while the control group taking a placebo of the formula. The treatment was administered once daily for 24 weeks. The primary outcomes included 24-hour urine total protein （24 h-UTP）， serum albumin （Alb）， glycated hemoglobin （HbA1c）， and serum creatinine （Scr）.The secondary outcomes included changes in levels of endothelin-1 （ET-1）， nitric oxide （NO）， vascular endothelial growth factor （VEGF）， and traditional Chinese medicine （TCM） syndrome scores before and after treatment. Clinical efficacy was evaluated based on TCM syndrome scores and overall disease outcomes. Adverse reactions and endpoint events were recorded. ResultsIn the treatment group after treatment， 24 h-UTP， ET-1， and VEGF levels significantly decreased （P＜0.05）， Alb and NO levels significantly increased （P＜0.05）； while the TCM syndrome scores for edema， lumbar pain， numbness of limbs， dark purple lips， dark purple tongue or purpura， and thin， rough pulse all significantly decreased （P＜0.05）. In the control group， no significant changes were observed in any of the indicators after treatment （P＞0.05）.Compared with the control group， the treatment group showed significant reductions in 24 h-UTP， ET-1， and VEGF levels， and increases in Alb and NO levels （P＜0.05）. The TCM syndrome scores for edema， lumbar pain， dark purple tongue or purpura， and thin， rough pulse were all lower in the treatment group than in the control group （P＜0.05）. The total effective rate of TCM syndrome in the treatment group was 59.09% （26/44）， and the overall clinical effective rate was 45.45% （20/44）. In the control group， these rates were 15.22% （7/46） and 8.7% （4/46）， respectively， with the treatment group showing significantly better outcomes （P＜0.05）. A total of 7 adverse events occurred across both groups， with no significant difference （P＞0.05）. No endpoint events occurred during the study. ConclusionOn the basis of conventional treatment of Western medicine， HTF can further reduce urinary protein levels and improve clinical symptoms in patients with mid-stage diabetic kidney disease of blood stasis syndrome. The mechanism may be related to its effects on endothelial function.

Objective:To evaluate the clinical equivalence between a domestic calcipotriol/betamethasone dipropionate ointment and the originator product in the treatment of stable plaque psoriasis.Methods:A multicenter, randomized, double-blind, three-arm, parallel-group, active- and placebo-controlled study was conducted, and 449 patients aged 18 - 65 years with stable plaque psoriasis were enrolled from 25 hospitals (such as the First Affiliated Hospital of China Medical University). Eligible patients had a baseline physician's global assessment (PGA) score of ≥ 3 points, baseline body surface area (BSA) involvement of 5% - 30%, and a target lesion psoriasis area and severity index (TL-PASI) for plaque elevation of ≥ 3 points. Participants were randomly assigned in a 2:2:1 ratio to the test group ( n = 179), reference group ( n = 180), and placebo group ( n = 90), and applied the domestic calcipotriol/betamethasone dipropionate ointment, originator product, and ointment base respectively, once daily in the evening for 4 weeks. Efficacy and safety were assessed at weeks 1, 2, and 4. The primary efficacy endpoints were the treatment success rates and clinical success rates in each group at week 4. The per-protocol set (PPS) was used for the primary efficacy analysis, and the intention-to-treat (ITT) set for supplementary efficacy analysis. Equivalence between the test and reference preparations was tested using the Cochran-Mantel-Haenszel method adjusted for randomization strata. Superiority of the test and reference preparations over the placebo was also tested. Measurement data were compared among the 3 groups using analysis of variance or non-parametric tests, while treatment success rates, clinical success rates, and incidence rates of adverse reactions were compared using the chi-square test. Results:The ITT, PPS, and safety sets included 447, 420, and 448 patients, respectively. In the ITT set, patients were aged 43.6 ± 12.8 years, including 320 (71.6%) males and 127 (28.4%) females, and the disease duration was 11.21 ± 9.05 years; 316 (70.7%) had a PGA score of 3 points and 131 (29.3%) had a PGA score of 4 - 5 points. No significant differences in the baseline characteristics (including age, sex, disease duration and disease severity) were observed among the 3 groups (all P > 0.05). Based on the PPS analysis, the treatment success rates were 57.9% (99/171) in the test group, 50.3% (86/171) in the reference group, and 7.7% (6/78) in the placebo group, and the clinical success rates were 57.9% (99/171), 50.3% (86/171), and 10.3% (8/78), respectively; both the test and reference groups were superior to the placebo group in both treatment and clinical success rates (all P < 0.001) ; the rate differences for treatment success (90% confidence interval [ CI]: -1.3% - 16.4%) and clinical success (90% CI: -1.3% - 16.3%) between the test and reference groups were entirely within the pre-defined equivalence margin (-20% - 20%). Subgroup analyses by baseline PGA scores: for patients with a baseline PGA score of 3 points, the treatment success rates in the test, reference, and placebo groups were 60.8% (73/120), 52.1% (62/119), and 11.1% (6/54), respectively, and the corresponding clinical success rates were 61.7% (74/120), 53.8% (64/119), and 13% (7/54), respectively; the test and reference groups did not differ significantly in treatment or clinical success rates (both P > 0.05), but both showed higher success rates than the placebo group (all P < 0.001) ; the results of statistical comparisons among the 3 groups in patients with a baseline PGA score of 4 - 5 points were consistent with those observed in patients with a baseline PGA score of 3 points. The percentage reductions in PGA and TL-PASI scores from baseline to weeks 1, 2, and 4 showed significant differences among the 3 groups, which were significantly higher in the test and reference groups than in the placebo group (all P < 0.001), but did not differ between the test and reference groups (all P > 0.05). The primary adverse reactions were local skin reactions, such as pruritus, pain, and erythema. The incidence rates of adverse reactions were 8.9% (16/179) in the test group, 7.3% (13/179) in the reference group, and 7.8% (7/90) in the placebo group, with no significant difference among the 3 groups ( P > 0.05) . Conclusions:The domestic calcipotriol/betamethasone dipropionate ointment demonstrated clinical equivalence to the originator product in the treatment of stable plaque psoriasis, and the two agents exhibited comparable efficacy for patients with varying degrees of disease severity, and were comparable in the speed and degree of clinical improvement, with similar favorable safety profiles.

Objective:To investigate the reliability and validity of the China Nanjing consciousness recovery scale(CNCRS)in assessing patients with acute disorder of consciousness(DOC).Method:A total of 306 patients with acute DOC admitted to Nanjing Zijin Hospital between August 2023 and August 2024 were enrolled.Each patient was assessed using both the Glasgow Coma Scale(GCS)and CNCRS.The correlation between GCS and CNCRS scores were analyzed,and the reliability and validity of these two scales were compared.Result:The DOC degree of enrolled patients was severe,with a tracheotomy rate of 61.8%.The median GCS score was 7(5,10)for males and 6(5,9)for females,while the median CNCRS score was 3(1,8)for males and 3(1,7)for females.Scores from both scales were statistically significant correlated across gender groups(P<0.001).The Cron-bach's α coefficient of CNCRS(male 0.918,female 0.889)was higher than that of GCS(male 0.687,female 0.658),indicating superior internal consistency.The validity index Kaiser-Meyer-Olkin(KMO)value(male 0.891,female 0.857)was also higher than the GCS scale(male 0.600,female 0.602),demonstrating better construct validity.Conclusion:The CNCRS demonstrates good reliability and validity and can be effectively applied for the evalu-ation of patients with acute disturbance of consciousness.

Objective To explore the neuroprotective effects and mechanisms of asiaticoside(AS)in rats with transient cerebral ischemia.Methods One hundred male Sprague-Dawley(SD)rats were randomly divided into five groups:sham-operated(Sham)group,transient middle cerebral artery occlusion(tMCAO)group,and low-,medium-,and high-dose AS(AS-L,AS-M,AS-H)groups,with 20 rats in each group.Except for the Sham group,rats in the other four groups under-went tMCAO surgery.Rats in the AS-L,AS-M,and AS-H groups received intragastric administration of 20,40 and 80 mg/kg AS respectively,once daily for 7 days starting 1 hour post-surgery.Rats in the Sham and tMCAO groups received equivalent volumes of saline.Neurological deficit score,brain water content,and TTC staining were used to evaluate neurological impairment,cerebral edema,and infarct volume.HE staining and Nissl staining wereused to assess histopathological changes and neu-ronal damage.Autophagy was detected via transmission electron microscopy.Immunofluorescence staining was usedto analyze the expression and localization of microtubule-associated protein light chain 3B(LC3B).TUNEL staining was used to evaluate apoptosis,and Western blot was used to measure protein expression.Results Compared with the Sham group,rats in the tMCAO group ex-hibited significantly increased neurological deficit score,brain water content,infarct volume,and histopathological damage,as well as significantly decreased Nissl body counts(P＜0.05).AS dose-dependently reduced neurological deficits,brain water content,infarct volume,and histopatho-logical damage while increased Nissl body numbers.The tMCAO group showed significantly higher numbers of autophagosomes,lysosomes,and LC3B-positive cells,along with significantly elevated LC3-Ⅱ/LC3-Ⅰ,Beclin-1,Bax,cleaved caspase-3 compared to the Sham group;in contrast,p-PI3K,p-Akt,and p-mTOR protein levels,intact mitochondria count and p62 and Bcl-2 protein levels were significantly lower inthe tMCAO group(P＜0.05).Compared with the tMCAO group,AS treatment dose-dependently significantly decreased autophagosomes,lysosomes,LC3B-positive cells,and the expression of LC3-Ⅱ/LC3-Ⅰ,Beclin-1,Bax,cleaved caspase-3 while significantly increased p-PI3K,p-Akt,and p-mTOR protein,intact mitochondria and p62 and Bcl-2 levels(P＜0.05).Conclusion AS exerts neuroprotective effects by inhibiting excessive autophagy and apopto-sis in rats with transient cerebral ischemia via activation of the PI3K/Akt/mTOR signaling pathway.

Objective To clarify the biomechanical properties of oblique lumbar interbody fusion(OLIF)combined with lateral plate(LP)fixation under different bone conditions by means of finite element(FE)analysis,so as to provide mechanical basis for clinical practice.Methods The three-dimensional nonlinear L3_5 FE model of the spine was constructed.Different material properties were assigned to each part of the model to establish a model of normal bone(NB),osteopenia(OS)and osteoporosis(OP)of the lumbar spine.OLIF combined with the following three internal fixation models were established.For NB,there were OLIF alone(M0),OLIF+LP fixation(M1),and OLIF+bilateral pedicle screw(BPS)fixation(M2);for OS,there were OLIF alone(N0),OLIF+LP fixation(N1),and OLIF+BPS fixation(N2);for OP,there were OLIF alone(P0),OLIF+LP fixation(P1),and OLIF+BPS fixation(P2).A 500 N load was applied on the upper surface of L3 to represent the weight of the upper body,and a 7.5 N·m moment was used to simulate the vertebral motion under six different conditions:flexion(FL),extension(EX),left bending(LB),right bending(RB),left rotation(LR)and right rotation(RR).The range of motion(ROM)of the fixed segment of NB,OS and OP models was calculated,and the overall stress of the vertebral body,the stress of the cage and the internal fixation device were recorded.Results Compared with the complete model,the stability of each surgical model increased.Compared with M0,ROM of M1 decreased,especially in the LB and RB,which was roughly equivalent to the stability of BPS but weaker than BPS in the control FL and EX direction.The situation in OS and OP model was similar to that in NB.Under the same internal fixation,L4_5 ROM gradually increased with bone deterioration.The overall intervertebral ROM of N1 in the FL direction increased by 16.1％compared with M1,P1 in the FL direction increased by 32.1％compared with M1,and P1 in the FL direction increased by 19.0％compared with N1.With the increase of posterior internal fixation,the overall stress of vertebral body,Cage and internal fixation stress showed a downward trend.Under the same internal fixation,with the loss of bone mass,the overall stress of the model gradually increased.Compared with M0,P0 increased the most in LR,reaching 56.5％.In the NB and OS models,the peak stress of the LP fixation under FL exceeded the minimum yield strength of the material by 22.7％and 33.8％,respectively,and was less than the minimum fatigue strength and yield strength of the material under the rest of the motion.In the OP,the peak stress of the internal fixation exceeded the minimum fatigue strength of the material at FL and EX,and reached 53.3％(＞50％)at FL.Conclusions Under NB and OS,OLIF combined with LP fixation can significantly improve the stability of the surgical segment,especially in LB and RB directions,and the overall stability is weaker than that of BPS fixation.Under OP,FL and EX may increase the risk of internal fixation failure.Combination with BPS fixation should be considered to improve the safety of fixation.

Objective:To analyze the clinical characteristics and cytokines expression characteristics in infants with mild and severe respiratory syncytial virus (RSV) infection.Methods:From May 2023 to December 2023, plasma samples and clinical information were collected from 16 infants with RSV infection and 14 control infants. Cytek Aurora flow cytometry (Cytek, America) and Enzyme linked immunosorbent assay (ELISA) were used to detect the expression levels of 25 cytokines after mild and severe RSV infection.Results:Cough and nasal obstruction were the main clinical manifestations in infants with mild RSV infection, accompanied by polypnea, wheezing and other symptoms. The main symptoms of severe RSV infection were cough and rales, accompanied by fever and polypnea. In comparison with the control group, the expression levels of IL-2, IL-4, IL-5, IL-6, IL-9, IL-13, IL-22, TNF-α, IFN-α, IFN-β, MIP-1β, I-TAC, ENA-78, GROα, Eotaxin, and MCP-1 in the RSV infection group all exhibited an upregulation trend. Both IP-10 and MIP-3α demonstrated a downward trend in the RSV infection group; however, there was no statistically significant difference ( P>0.05). The levels of IL-10, IFN-γ, MIP-1α, and IL-8 in the RSV infection group were significantly higher than those in the control group, whereas the levels of MIG, TARC, and RANTES in the RSV infection group were significantly lower than those in the control group ( P<0.05). The levels of IL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-13, IL-22, IFN-β, IFN-γ, TNF-α, IL-8, I-TAC, MIP-1β, Eotaxin, and MCP-1 in the mild RSV infection group were significantly higher than those in the severe RSV infection group ( P>0.05). Among these, the levels of MIG, RANTES, TARC, MIP-3α, and ENA-78 in the mild infection group were all lower than those in the severe infection group. The expressions of ENA-78 and MIP-1α in the severe infection group were significantly higher than those in the mild infection group and also higher than those in the control group. There was no significant difference in IP-10 and GROα between the mild and severe RSV infection groups ( P>0.05). Conclusions:The differences in clinical features and cytokines between infants with mild and severe RSV infection provide important data support for the prevention and treatment of RSV infection in infants.

Objective To explore the efficacy of vacuum-assisted minimally invasive gyrotomy for benign lesions in the lower quadrant of the breast.Methods From August 2022 to August 2023,90 patients with benign lesions in the lower quadrant of the breast were treated.According to the different surgical methods,they were divided into two groups:the traditional group included 45 cases,which underwent surgery through the trans-areolar incision;the minimally invasive group included 45 cases,which underwent surgery through the trans-mammary fold incision with vacuum-assisted minimally invasive technique.The surgical-related indicators,inflammatory response indicators,patient satisfaction,and safety were compared between the two groups.Results The operation time[(20.45±6.18)min],intraoperative bleeding[(7.78±2.23)ml],hospital stay[(4.37±1.05)d],scar length[(2.32±0.42)cm]and healing time[(3.46±1.08)d]in the minimally invasive group were significantly better than those in the traditional group[(34.52±9.46)min,(23.16±6.44)ml,(8.72±2.73)d,(19.14±4.18)cm and(7.37±2.16)d](P＜0.05).The levels of IL-6[(12.14±2.86)ng/L],IL-10[(14.33±3.74)pg/ml],CRP[(13.85±3.11)mg/L],Cor[(131.27±6.43)nmol/L]and NE[(82.55±8.44)in the minimally invasive group at 3 days after surgery ng/ml were increased and VEGF was decreased[(59.72±7.44)mg/L]than the traditional group[(17.23±3.38)ng/L,(19.62±4.88)pg/ml,(28.36±4.67)mg/L,(196.52±8.84)nmol/L,(117.62±7.14)ng/ml and(88.46±7.89)mg/L](P＜0.05).The satisfaction of patients in minimally invasive group(97.78％,44/45)was significantly higher than that in traditional group(82.22％,37/45)(P＜0.05).The incidence of adverse events in minimally invasive group(4.44％,2/45)was significantly lower than that in traditional group(22.22％,10/45)(P＜0.05).Conclusion Vacuum-assisted minimally invasive rotatory excision can optimize surgical indicators,improve serological indicators,enhance patient satisfaction and treatment safety compared to traditional areola incision surgery.