Objective:To investigate the effectiveness and safety of low-dose oral misoprostol solution for cervical ripening in late gestation.Methods:This was a prospective cohort study including 396 primiparas with singleton pregnancy who received low-dose oral misoprostol solution for cervical ripening (oral group) in Peking University Third Hospital from March to October 2022. They were further allocated to receive oral misoprostol alone (OA group, n=167) or oral misoprostol in combination with oxytocin/amniotomy (OC group, n=229). Moreover, 218 cases who received vaginal misoprostol for cervical ripening (vaginal group) during the same period in 2021 were reviewed (a retrospective cohort). Among them, 77 were given vaginal misoprostol alone (VA group) and 141 received vaginal misoprostol in combination with oxytocin/amniotomy (VC group). The OA group and VA group (72 and 73 cases) as well as the OC group and VC group (108 and 103 cases) were matched using propensity scores. Basic clinical information, hospital stay, duration of labor induction, uterine hyperstimulation, rate of labor initiation, vaginal delivery rate, rate of delivery within 24 h, duration of labor, neonatal condition, adverse pregnancy outcomes, and other information were compared between different groups. All data were statistically analyzed using independent sample t test, analysis of variance, nonparametric test, Chi-square test, or Fisher's exact probability test. Logistic regression model was used to analyze the factors affecting the labor initiation and the failure of labor induction. Results:The average hospital stay, the duration from medication to labor initiation and the duration from medication to vaginal delivery were significantly shorter in the oral group than those in the vaginal group [(5.4±2.4) vs. (6.5±2.6) d, (34.2±24.1) vs. (38.9±25.7) h, (45.8±25.8) vs. (53.4±27.8) h; t=5.24, 2.10 and 3.39; all P<0.05]. The total labor initiation rate and vaginal delivery rate in the oral group were significantly higher than those in the vaginal group [92.9% (368/396) vs. 83.5% (182/218), 72.2% (286/396) vs. 60.1% (131/218); χ 2=13.43 and 9.50; both P<0.05]. The incidence of failed induction of labor, uterine hyperstimulation, fetal distress, and intrauterine infection in the oral group were lower than those in the vaginal group [2.0% (8/396) vs. 6.9% (15/218), 4.3% (17/396) vs. 17.9% (39/218), 8.8% (35/396) vs. 14.7% (32/218), 1.3% (5/396) vs. 3.7% (8/218); χ 2=9.21, 31.36, 4.93 and 3.93; all P<0.05]. The duration from medication to labor initiation and to vaginal delivery in the OA group were higher than those in the VA group [(25.8±17.0) vs. (17.4±10.8) h, (37.2±18.8) vs. (29.7±13.5) h; t=3.49 and 2.74; both P<0.05]. There were no significant differences in the labor initiation rate, vaginal delivery rate, rate of delivery within 24 h or the incidence of failed induction of labor between the OA and VA groups (all P>0.05). Women in the VA group were more likely to develop uterine hyperstimulation than those in the OA group [19.2% (14/73) vs. 4.2% (3/72), χ2=7.89, P=0.005]. There were no significant differences in the duration from medication to labor initiation or to vaginal delivery between the VC and OC groups (both P>0.05), but the duration were significantly longer than those in the corresponding medication alone group (VC vs. VA groups: (49.7±24.6) vs. (17.4±10.8) h and (61.6±25.7) vs. (29.7±13.5) h, t=5.31 and 5.13, both P<0.05; OC vs. OA groups: (45.3±26.6) vs. (25.8±17.0) h and (56.1±27.2) vs. (37.2±18.8) h, t=10.35 and 9.78, both P<0.05]. The labor initiation rate, vaginal delivery rate and rate of delivery within 24 h in the OC group were higher than those in the VC group [88.9% (96/108) vs. 77% (87/113), 63.0% (68/108) vs. 47.8% (54/113), 10.3% (7/108) vs. 0.0% (0/113); χ 2=5.49, 5.14 and 7.56; all P<0.05]. The incidence of uterine hyperstimulation in the OC group was 4.6% (5/108), which was lower than that in the VC group [18.6% (21/113), χ 2=10.37, P=0.001]. Logistic regression analysis showed that oral misoprostol and gestational age were positively correlated with labor initiation [ OR (95% CI): 2.18 (1.24-3.90) and 1.43 (1.14-1.79)], while maternal age was negatively correlated with labor initiation [ OR (95% CI): 0.90 (0.82-0.98)]. Moreover, failed induction of labor was negatively correlated with oral misoprostol [ OR (95% CI): 0.37 (0.14-0.91)], but positively correlated with maternal age [ OR (95% CI): 1.21 (1.05-1.40)]. Conclusions:Oral administration of low-dose misoprostol solution is as effective as vaginal misoprostol in promoting cervical ripening. Besides, it can shorten the average hospital stay and reduce the incidence of uterine hyperstimulation, suggesting that low-dose oral misoprostol solution is relatively safer and can be used to promote cervical ripening in late gestation.

BACKGROUND:It is known that N6-methyladenosine(m6A)plays a role in the pathogenesis of various diseases and studies have suggested its involvement in the pathologic changes of steroid-induced femoral head necrosis(SNFH).However,research on m6A methylation modifications in steroid-induced femoral head necrosis is limited. OBJECTIVE:Using bioinformatics methods to identify the differential expression of m6A-related genes in steroid-induced femoral head necrosis and to predict miRNAs associated with these genes to further elucidate the role and mechanism of m6A methylation in steroid-induced femoral head necrosis. METHODS:Differential gene expression between steroid-induced femoral head necrosis and control groups was analyzed using GSE123568 gene expression data and identified using the"limma"package in R.Functional enrichment analysis was performed on the differentially expressed genes.Differential analysis of the related genes was carried out using the"ggstatsplot"package in R.The differential genes were cross-validated using the GSE74089 dataset.An mRNA-miRNA regulatory network was constructed,and co-expression analysis was performed on the module genes followed by enrichment analysis.Differences in immune cell infiltration between steroid-induced femoral head necrosis and control groups were quantified using the ssGSEA method. RESULTS AND CONCLUSION:Correlation analysis revealed 13 m6A-related genes,and further analysis through the protein-protein interaction network identification and receiver operating characteristic curve analysis showed that YTHDF2 was expected to be a core differential gene as a potential early biomarker.Enrichment analysis indicated that differentially expressed genes were mainly involved in inflammation and immune response and were closely related to osteoclasts.Cross-validation analysis showed that differential gene expression results between the two datasets were consistent.mRNA-miRNA regulatory network analysis revealed that YTHDF2 was negatively correlated with miRNA-27a.Immune infiltration analysis revealed an increase in immune cell infiltration in steroid-induced femoral head necrosis,and YTHDF2 was positively correlated with the infiltration of CD4+T cells.To conclude,m6A-related gene YTHDF2 can serve as a potential biomarker of steroid-induced femoral head necrosis and is valuable for the early clinical diagnosis and treatment of steroid-induced femoral head necrosis.The negative correlation between YTHDF2 and mir-27a and the positive correlation between YTHDF2 and CD4+T cell infiltration provide new insights into the early diagnosis and treatment of steroid-induced femoral head necrosis and shed light on the mechanism of m6A in steroid-induced femoral head necrosis.

Objective:To compare the effects of leukocyte-poor platelet-rich plasma (Lp-PRP) and leukocyte-rich platelet-rich plasma (Lr-PRP) on dorsal wound healing in rats.Methods:Thirty-six male Sprague-Dawley rats were randomly divided into Lp-PRP group, Lr-PRP group and control group, each containing twelve rats. Venous blood was drawn and the Lp-PRP and Lr-PRP were prepared separately using a centrifugal method. Circular full-thickness skin defect wounds (15 mm in diameter) were created on the backs of the rats in the three groups. The wounds were then treated with 100 μl Lp-PRP, Lr-PRP and saline, respectively. At 7 and 14 days post-operation, the wounds were photographed, and Image J software was used to calculate the wound area rate (postoperative wound area/wound area at modeling time × 100%). At 14 days post-operation, the total neo-epithelium length and collagen deposition rate of the wounds were evaluated using HE and Masson staining, respectively. At 7 days post-operation, the relative expression of vascular endothelial growth factor (VEGF) in the wounds was detected by Western blotting, and the number of CD31 positive microvessels in the wounds was examined by immunohistochemistry. Statistical analysis was performed using SPSS 28.0. One-way analysis of variance (ANOVA) was used to compare the three groups, and Tukey’s test was used for pairwise comparisons. A significance level of P<0.05 was considered statistically significant. Results:Blood analysis revealed that the platelet concentrations in the prepared Lp-PRP and Lr-PRP were 4.1 times and 4.5 times that of whole blood, respectively ( P<0.01), with no significant difference between the two PRPs ( P>0.05). The leukocyte concentration in Lp-PRP was undetectable, while in Lr-PRP, it was 3.5 times that of whole blood ( P<0.01). The wound area rate at 7 and 14 days post-operation, the total neo-epithelium length and collagen deposition rate at 14 days post-operation, as well as the relative expression of VEGF and the number of CD31-positive microvessels at 7 days post-operation in the Lp-PRP and Lr-PRP groups were superior to those in the control group (all P<0.01). However, there was no significant difference between the two PRP groups (all P>0.05). Conclusion:Both Lp-PRP and Lr-PRP promote dorsal wound healing in rats by enhancing re-epithelialization, collagen deposition, and angiogenesis. The impacts of Lp-PRP and Lr-PRP on promoting wound healing are comparable and not influenced by the presence of leukocytes in PRPs.

Objective:To compare the effects of leukocyte-poor platelet-rich plasma (Lp-PRP) and leukocyte-rich platelet-rich plasma (Lr-PRP) on dorsal wound healing in rats.Methods:Thirty-six male Sprague-Dawley rats were randomly divided into Lp-PRP group, Lr-PRP group and control group, each containing twelve rats. Venous blood was drawn and the Lp-PRP and Lr-PRP were prepared separately using a centrifugal method. Circular full-thickness skin defect wounds (15 mm in diameter) were created on the backs of the rats in the three groups. The wounds were then treated with 100 μl Lp-PRP, Lr-PRP and saline, respectively. At 7 and 14 days post-operation, the wounds were photographed, and Image J software was used to calculate the wound area rate (postoperative wound area/wound area at modeling time × 100%). At 14 days post-operation, the total neo-epithelium length and collagen deposition rate of the wounds were evaluated using HE and Masson staining, respectively. At 7 days post-operation, the relative expression of vascular endothelial growth factor (VEGF) in the wounds was detected by Western blotting, and the number of CD31 positive microvessels in the wounds was examined by immunohistochemistry. Statistical analysis was performed using SPSS 28.0. One-way analysis of variance (ANOVA) was used to compare the three groups, and Tukey’s test was used for pairwise comparisons. A significance level of P<0.05 was considered statistically significant. Results:Blood analysis revealed that the platelet concentrations in the prepared Lp-PRP and Lr-PRP were 4.1 times and 4.5 times that of whole blood, respectively ( P<0.01), with no significant difference between the two PRPs ( P>0.05). The leukocyte concentration in Lp-PRP was undetectable, while in Lr-PRP, it was 3.5 times that of whole blood ( P<0.01). The wound area rate at 7 and 14 days post-operation, the total neo-epithelium length and collagen deposition rate at 14 days post-operation, as well as the relative expression of VEGF and the number of CD31-positive microvessels at 7 days post-operation in the Lp-PRP and Lr-PRP groups were superior to those in the control group (all P<0.01). However, there was no significant difference between the two PRP groups (all P>0.05). Conclusion:Both Lp-PRP and Lr-PRP promote dorsal wound healing in rats by enhancing re-epithelialization, collagen deposition, and angiogenesis. The impacts of Lp-PRP and Lr-PRP on promoting wound healing are comparable and not influenced by the presence of leukocytes in PRPs.

Objective:To investigate the clinical efficacy of over-the-top reconstruction combined with the modified Lemaire technique in the treatment of anterior cruciate ligament (ACL) injuries with pivot-shift positive.Methods:From March 2020 to October 2021, a total of 46 patients with ACL injury and pivot-shift test grade II or above were admitted to Xinhua Hospital Affiliated to Dalian University. There were 28 males and 18 females, aged 28.0±10.5 years (range, 15-45 years). All cases were unilateral, including 17 cases of left knee and 29 cases of right knee. The pivot-shift test showed that 30 cases were grade II and 16 cases were grade III, and the cause of injury was sports injury. The semitendinosus muscle and gracilis muscle were harvested, and the ACL was reconstructed with the over-the-top combined modified Lemaire technique. The International Knee Documentation Committee (IKDC) score, Lysholm score and KT-2000 side-to-side difference before and after operation were compared.Results:All patients successfully completed the operation and were followed up for 26.6±2.3 months (range, 24-28 months). The Lysholm scores of the patients at 3 months and 24 months after operation were 73.6±4.3 and 91.6±2.8, which were higher than those before operation (58.5±4.6), and the difference was statistically significant ( F=18.351, P<0.001). The IKDC scores of patients at 3 months and 24 months after operation were 59.0±2.0 and 91.8±3.2, respectively, which were higher than those before operation 50.3±2.8, and the difference was statistically significant ( F=17.290, P<0.001). The side-to-side difference of KT-2000 was 1.7±0.8 mm and 1.5±0.4 mm at 3 and 24 months after operation, respectively, which was lower than that before operation (5.9±1.1 mm), and the difference was statistically significant ( F=14.192, P<0.001). At 24 months after operation, 3 patients had pivot shift test grade I and 4 patients had Lachman test grade I, but they complained of good knee stability and did not receive further treatment. At the last follow-up, there were no complications such as incision and intra-articular infection, deep vein thrombosis, knee stiffness, quadriceps musculus ossificans myositis, and reconstruction ligament rupture. All patients returned to sports with an average time of 15.7±2.6 months (range, 12-24 months). Conclusion:Over-the-top reconstruction combined with the modified Lemaire technique for the treatment of ACL injury with positive pivot shift test effectively improves knee function and promotes the patient's return to sports, with a low incidence of surgical complications.

Objective:To evaluate the safety and efficacy of enhanced recovery after surgery(ERAS) in the perioperative period of congenital choledochal cysts in children.Methods:This is a multicenter prospective randomized controlled study. The clinical data of 273 pediatric congenital choledochal cysts(CCC) patients who underwent surgery at 14 medical centers with complete follow-up data were collected through the medical data analysis platform. Among them, 123 cases in ERAS group were managed perioperatively in strict accordance with ERAS mode, and 150 cases in conventional group were managed according to traditional mode. The length of hospital stay,time to first farting, time to complete feeding, the incidence of complications, cost and readmission rate within 30 days,stress indexes and liver function were compared between the two groups.Results:Compared with the conventional group, median time to start farting (2.0 d vs. 3.0 d, P<0.001), median time to complete feeding (5.0 d vs. 7.0 d, P<0.001), median postoperative hospitalization time (6.0 d vs. 9.0 d, P<0.001),the median total length of stay(13.0 d vs. 15.0 d, P<0.001) were shorter,the median hospitalization cost (37,000 yuan vs.43,000 yuan P<0.001) was lower, and stress indexes recovered quickly. The incidence of postoperative hospital stay and readimission rate within 30 d were not statistically different between the two groups. Conclusion:It is safe and feasible to implement ERAS for children with CCC in the perioperative period, which can reduce stress response, speed up recovery,and save medical costs.

Marsdenia tenacissima injection, a standard Marsdenia tenacissima extract (MTE), has been approved as an adjuvant therapeutic agent for various cancers. Our previous study showed that MTE inhibited the proliferation and metastasis of prostate cancer (PCa) cells. However, the underlying mechanisms and active ingredients of MTE against PCa were not completely understood. This study revealed that MTE induced significant decreases in cell viability and clonal growth in PCa cells. In addition, MTE induced the apoptosis of DU145 cells by reducing the mitochondrial membrane potential and increasing the expression of Cleaved Caspase 3/7, Cyt c, and Bax. In vivo, DU145 xenografted NOD-SCID mice treated with MTE showed significantly decreased tumor size. TUNEL staining and Western blot confirmed the pro-apoptotic effects of MTE. Network pharmacology analysis collected 196 ingredients of MTE linked to 655 potential targets, and 709 PCa-associated targets were retrieved, from which 149 overlapped targets were screened out. Pathway enrichment analysis showed that the HIF-1, PI3K-AKT, and ErbB signaling pathways were closely related to tumor apoptosis. Western blot results confirmed that MTE increased the expression of p-AKT^Ser473 and p-GSK3β^Ser9, and decreased the expression of p-STAT3^Tyr705in vitro and in vivo. A total of 13 compounds in MTE were identified by HPLC-CAD-QTOF-MS/MS and UPLC-QTOF-MS/MS. Molecular docking analysis indicated that six compounds may interact with AKT, GSK3β, and STAT3. In conclusion, MTE induces the endogenous mitochondrial apoptosis of PCa by regulating the AKT/GSK3β/STAT3 signaling axis, resulting in inhibition of PCa growth in vitro and in vivo.
Mice ; Animals ; Male ; Humans ; Mice, Inbred NOD ; Mice, SCID ; Marsdenia ; Proto-Oncogene Proteins c-akt ; Glycogen Synthase Kinase 3 beta ; Molecular Docking Simulation ; Phosphatidylinositol 3-Kinases ; Tandem Mass Spectrometry ; Prostatic Neoplasms ; Apoptosis ; STAT3 Transcription Factor

Background: The emergence of the severe acute respiratory syndrome coronavirus 2 omicron variant has been triggering the new wave of coronavirus disease 2019 (COVID-19) globally. However, the risk factors and outcomes for radiological abnormalities in the early convalescent stage (1 month after diagnosis) of omicron infected patients are still unknown. Methods: Patients were retrospectively enrolled if they were admitted to the hospital due to COVID-19. The chest computed tomography (CT) images and clinical data obtained at baseline (at the time of the first CT image that showed abnormalities after diagnosis) and 1 month after diagnosis were longitudinally analyzed. Uni-/multi-variable logistic regression tests were performed to explore independent risk factors for radiological abnormalities at baseline and residual pulmonary abnormalities after 1 month. Results: We assessed 316 COVID-19 patients, including 47% with radiological abnormalities at baseline and 23% with residual pulmonary abnormalities at 1-month follow-up. In a multivariate regression analysis, age ≥ 50 years, body mass index ≥ 23.87, days after vaccination ≥ 81 days, lymphocyte count ≤ 1.21 × 10 -9 /L, interleukin-6 (IL-6) ≥ 10.05 pg/mL and IgG ≤ 14.140 S/CO were independent risk factors for CT abnormalities at baseline. The age ≥ 47 years, presence of interlobular septal thickening and IL-6 ≥ 5.85 pg/mL were the independent risk factors for residual pulmonary abnormalities at 1-month follow-up. For residual abnormalities group, the patients with less consolidations and more parenchymal bands at baseline could progress on CT score after 1 month. There were no significant changes in the number of involved lung lobes and total CT score during the early convalescent stage. Conclusion The higher IL-6 level was a common independent risk factor for CT abnormalities at baseline and residual pulmonary abnormalities at 1-month follow-up. There were no obvious radiographic changes during the early convalescent stage in patients with residual pulmonary abnormalities.

Objective:To explore the protective effect of methyl eugenol (Me) on islet ischemia/reperfusion (I/R) injury and elucidate its underlying mechanism.Methods:The islets were isolated and purified from 6-8 week male BALB/c mice and divided into four groups of normal control (normal culture without any treatment), hypoxia/reoxygenation (H/R treatment), H/R+ dimethyl sulfoxide (DMSO dosing plus H/R treatment) and H/R+ Me (Me dosing plus H/R). Viability of islet cells in each group was detected by acridine orange (AO)/propidium iodide (PI) double stain.Function of islet cells (insulin secretion) was measured by enzyme-linked immunosorbent assay (ELISA). Murine islet β Min6 cells were selected for detecting the effect of Me on the proliferative activity of normal cultured and H/R treated islet cells under different concentration gradients by CCK8.Then Min6 cells were divided into four groups of normal, H/R, H/R+ DMSO and H/R+ Me.The definition of group was the same as that of primary murine islets.Flow cytometry and Hoechst 33342 nuclear stain were utilized for detecting cell apoptotic rate in each group.The protein expressions of p-JNK, p-p38, JNK, p38, Bcl-2 and Bax were detected by Western blot.And the data were processed by one-way ANOVA or t test.Results:The proportion of dead islet cells in H/R group was (29.47±2.65)% and it was significantly lower than that in normal group (7.63±1.53)%.And the inter-group differences were statistically significant ( P<0.001). The proportion of dead islet cells was (20.63±3.07)% in H/R+ Me group.It was higher than that in H/R group (29.47±2.65)% and in H/R+ DMSO group (30.13±1.50)% and inter-group difference was statistically significant ( P<0.05 & P<0.01). Under the stimulation of high glucose, the insulin secretion level of islet in H/R+ Me group was (1.76+ 0.08) mg/L, which was higher than that in H/R group and H/R+ DMSD group(1.24±0.14)mg/L and(1.27±0.05)mg/L, and the difference was statistically significant[(1.76±0.08) vs. (1.24±0.14) mg/L; (1.76±0.08) vs.(1.27±0.05) mg/L, P<0.01]. There was no significant effect on cell viability after Me dosing within a certain concentration range (0-40 μmol/L). After Me dosing (5 μmol/L), cell viability of H/R-treated Min6 cells was significantly higher than that without Me.And the difference was statistically significant[(1.19±0.03) vs.(1.00±0), P<0.01]. As compared with H/R and H/R+ DMSO groups, overall apoptotic rate declined in H/R+ Me group (Hoechst 33342 stain: 14.50%±1.05% vs. 23.30%±1.18%, 14.50%±1.05% vs. 22.77%±1.75%, P<0.001; Flow cytometry: 4.36%±0.54% vs. 21.44%±1.02%, 4.36%±0.54% vs. 21.68%±3.06%, P<0.01). The expressions of p-JNK and p-p38 were down-regulated (p-JNK: 0.77±0.06 vs. 1.03±0.05, 0.77±0.06 vs.0.93±0.04, P<0.001; p-p38: 0.80±0.05 vs. 1.01±0.08; 0.80±0.05 vs. 1.00±0.05, P<0.05) while Bcl-2/Bax ratio rose (1.62±0.13 vs. 0.72±0.10, 1.62±0.13 vs. 0.74±0.13, P<0.01). Conclusions:Me can improve the viability and function of islets and suppress the apoptosis of Min6 cells after H/R.The mechanism is correlated with JNK and p38 MAPK signaling pathways.

Geobacillus thermoglucosidasius is a kind of Gram-positive facultative anaerobic bacteria. The fast growth rate under high temperature and less susceptibility to microbial contamination enable G. thermoglucosidasius to be a desirable producer of biofuels and high-value-added chemicals for the next-generation industrial biotechnology. However, compared with the classical model strain Escherichia coli, the applications of G. thermoglucosidasius are hampered by its low transformation efficiency. This study aimed at obtaining competent cells with high transformation efficiency through inactivating restriction enzymes, adding cell membrane inhibitors and cell wall weakening agents. The results showed that the electro-transformation efficiency achieved 1.2×10⁴ CFU/(μg DNA) by knocking out four genes encoding restriction enzymes. Adding a certain amount of tween 80, dl-threonine and glycine further increased the competent efficiency about 22.5, 44, and 334 times, respectively. The electro-transformation efficiency was enhanced to 4.6×10⁶ CFU/(μg DNA) under the optimized conditions, laying a foundation for genetic manipulation and metabolic engineering of G. thermoglucosidasius.
Electroporation ; Electroporation Therapies ; Bacillaceae ; Cell Membrane ; Escherichia coli/genetics*