ObjectiveTo investigate the incidence rate of post-endoscopic retrograde cholangiopancreatography （ERCP） pancreatitis （PEP） in patients with difficult common bile duct intubation undergoing pancreatic duct stenting during surgery， as well as the effect of pancreatic duct stenting in the prevention and treatment of PEP， and to provide a basis for clinical treatment. MethodsA retrospective analysis was performed for the clinical data of 186 patients with biliary tract disease who underwent initial ERCP and had difficult common bile duct intubation in The First Affiliated Hospital of Zhengzhou University from January 2016 to December 2024， and according to the condition of pancreatic duct stenting， the patients were divided into control group with 73 patients （without pancreatic duct stenting）， 5Fr-5 cm stent group with 67 patients， and 7Fr-5 cm stent group with 46 patients. The three groups were compared in terms of baseline data， intraoperative procedures， and postoperative outcomes. A one-way analysis of variance was used for comparison of normally distributed continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups； the Kruskal-Wallis H rank sum test was used for comparison of non-normally distributed continuous data between multiple groups， and the Dunn method was used for further comparison between two groups； the chi-square test or the Fisher’s exact test was used for comparison of categorical data between groups. The Logistic regression analysis was used to investigate the influencing factors for PEP in patients with difficult intubation during ERCP. ResultsThe overall incidence rate of PEP was 12.37% （23/186）. Compared with the 5Fr-5 cm stent group and the 7Fr-5 cm stent group， the control group had a significantly higher incidence rate of PEP， a significantly higher score of postoperative abdominal pain， and a significantly longer length of postoperative hospital stay （all P0.01）， and 55.56% of the patients in the control group had moderate-to-severe PEP. The univariate Logistic regression analysis showed that intradiverticular papilla， double guide wire intubation， needle knife precut， the application of basket and balloon for removal of common bile duct stones， intraoperative biopsy， pancreatic duct stenting， intubation time≤10 minutes， frequency of intubation≤5 times， preoperative CRP≤5 mg/L were influencing factors for PEP （all P0.05）， and the multivariate Logistic regression analysis showed that intraoperative pancreatic duct stenting， needle knife precut， and intraoperative biopsy were independent influencing factors for the onset of PEP （all P0.05）. ConclusionPancreatic duct stenting during ERCP can effectively reduce the risk of PEP in patients with difficult intubation， while needle knife precut and intraoperative biopsy can increase the risk of PEP in patients with difficult intubation.

Bone morphogenetic proteins (BMPs) are multifunctional growth factors of the transforming growth factor β (TGF-β) superfamily. They regulate steroid secretion from mammalian granulosa cells, promote granulosa cell survival and proliferation, and inhibit follicular atresia, luteinization, and granulosa cell apoptosis, thereby promoting the development and maturation of mammalian follicles. At the same time, BMPs play an important role in embryonic morphogenesis, induction of uterine receptivity, and blastocyst attachment. This paper describes the effects of BMPs on mammalian follicular and embryonic development and the roles of BMPs in female reproduction, focusing on the process in which BMPs promote follicular maturation by regulating steroid secretion from granulosa cells during mammalian oocyte maturation. This review aims to provide a reference for further research on mammalian oocyte culture and improvement of reproductive efficiency in female animals.
Animals ; Embryonic Development/drug effects* ; Female ; Bone Morphogenetic Proteins/pharmacology* ; Reproduction/physiology* ; Humans ; Granulosa Cells/cytology* ; Oocytes

Objective To apply bedside ultrasound for real-time monitoring of intracranial conditions in patients with severe traumatic brain injury experiencing acute encephalocele during large craniotomy， and to investigate the clinical value of intraoperative bedside ultrasound in the diagnosis and prognostic evaluation of acute encephalocele. Methods A retrospective analysis was performed for 32 adult patients with severe traumatic brain injury， and according to whether intraoperative ultrasound was performed， they were divided into ultrasound group with 17 patients and CT group with 15 patients. The two groups were compared in terms of time of operation， accuracy， mortality rate， and postoperative Glasgow Outcome Scale （GOS） score. The chi-square test was used for comparison of categorical data between groups， and the independent samples t-test was used for comparison of continuous data between groups. Results Both ultrasound and CT could provide an accurate basis for diagnosis， with no significant difference in diagnostic accuracy； however， compared with the CT group， the ultrasound group had significantly shorter time of operation and diagnostic time. Based on GOS score and grading results at 6 months after surgery， the patients undergoing ultrasound examination had a significantly better prognosis than those undergoing CT examination. Conclusion Intraoperative ultrasound for patients with severe traumatic brain injury enables rapid and accurate identification of etiology， facilitates dynamic intracranial monitoring， and shortens the time for rescue， showing an important clinical significance in improving prognosis and reducing mortality rate. Therefore， it holds promise for clinical application.

Objective:To study the in vitro construction of functional and self-renewing cartilage organoids based on cartilage acellular extracellular matrix (ECM) microcarriers.Methods:Fresh porcine articular cartilage was taken. The merely crushed cartilage particles were set as natural cartilage group and ECM microcarriers of appropriate particle size, which were prepared by the acellular method of combining physical centrifugation and chemical extraction, were set as microcarrier group. Cartilage organoids were constructed by loading human umbilical cord mesenchymal stem cells (hUCMSCs) and human chondrocytes (hCho) with a ratio of 3∶1 with microcarriers through a rotating bioreactor. The organoids with different induction times were divided into 0-, 7-, 14-, and 21-day induction groups. The cell residues of the microcarrier group and natural cartilage group were evaluated by 4′, 6-diaminidine 2-phenylindole (DAPI) fluorescence staining and DNA quantitative analysis. The retention of microcarrier components was observed by Safranin O and toluidine blue stainnings, and the collagen and glycosaminoglycan (GAGs) levels in the microcarrier group and the natural cartilage group were determined by colorimetric method and dimethyl-methylene blue (DMMB) method. The microcarriers were further characterized by scanning electron microscopy and energy dispersive spectroscopy. The hUCMSCs cultured with Dulbecco′s Modified Eagle′s Medium (DMEM) supplemented with fetal bovine serum (FBS) in a volume fraction of 10% was used as the control group and the hUCMSCs cultured with the microcarrier extract was used as the experimental group. Subgroups of hUCMSCs cultured at 3 time points: 1, 3 and 5 days were set up in the two groups separately. Cell Counting Kit 8 (CCK-8) was used to detect the biocompatibility of the two groups. The cellular activity of the organoids of the 0-, 7-, 14-, and 21-day induction groups was detected by live/dead staining and the self-renewal ability of the cartilage organoids of the 14-day induced group was identified by Ki67 fluorescence staining. The organoids of the 7-, 14-, and 21-day induction groups were detected by RT-PCR in terms of the expression levels of chondrogenesis-related marker aggrecan (ACAN), type II collagen (COL2A1), SRY-related high mobility group-box gene-9 (SOX9), cartilage hypertrophy-and mineralization-related marker type I collagen (COL1A1), Runt-related transcription factor-2 (RUNX2), and osteocalcin (OCN). Colorimetric and DMMB assays were performed to determine the ability of organoids in the 0-, 7-, 14-, and 21-day induction groups to secrete collagen and GAGs.Results:The results of DAPI fluorescent staining showed that the natural cartilage group had a large number of nuclei while the microcarrier group hardly had any nuclei. The DNA content of the microcarrier group was (7.8±1.8)ng/mg, which was significantly lower than that of the natural cartilage group [(526.7±14.7)ng/mg] ( P<0.01). Saffranin O and toluidine blue staining showed that the microcarrier was dark- and uniform-colored and it kept a lot of cartilage ECM components. The collagen and GAGs contents of the microcarrier group were (252.9±1.4)μg/mg and (173.4±0.8)μg/mg, which were significantly lower than those of the natural cartilage group [(311.9±2.2)μg/mg and (241.3±0.7)μg/mg] ( P<0.01). Scanning electron microscopy showed that the surface of the microcarriers had uneven and interleaved collagen fiber network. The results of energy spectrum analysis showed that elements C, O and N were evenly distributed in the microcarriers, indicating that the composition of the microcarrier was uniform. The microcarrier had good biocompatibility and there was no statistical significance in the results of CCK-8 test between the control group and the experimental group after 1 and 3 days of culture ( P>0.05). After 5 days of culture, the A value of the experimental group was 0.53±0.02, which was better than that of the control group (0.44±0.03) ( P<0.05). In the 0-, 7-, 14-, and 21-day induction groups, hUCMSCs and hCho were attached to the surface of the microcarriers, with good cellular activity, and the live/death rates were (70.6±1.1)%, (80.5±0.6)%, (94.5±0.9)%, and (90.8±0.5)% respectively ( P<0.01). There were a large number of Ki67 positive cells in cartilage organoids. RT-PCR showed that the expression levels of ACAN, COL2A1, SOX9, COL1A1, RUNX2 and OCN were 1.00±0.09, 1.00±0.24, 1.00±0.18, 1.00±0.03, 1.00±0.06 and 1.00±0.13 respectively in the 7-day induction group; 4.16±0.28, 5.09±1.25, 5.65±1.05, 0.47±0.01, 1.68±0.02 and 0.21±0.06 respectively in the 14-day induction group; 13.42±0.92, 3.07±0.21, 1.84±1.08, 2.72±0.17, 2.91±0.18 and 3.32±1.20 respectively in the 21-day induction group. Compared with the 7-day induction group, the expression levels of ACAN, COL2A1, SOX9 and RUNX2 in the 14-day group were increased ( P<0.05), but COL1A1 expression level was decreased ( P<0.05), with no significant difference in OCN expression level ( P>0.05). Compared with the 7-day induction group, the expression levels of ACAN, COL1A1 and RUNX2 in the 21-day induction group were significantly increased ( P<0.01), with no significant differences in the expression levels of COL2A1, SOX9 and OCN ( P>0.05). Compared with the 14-day induction group, the expression levels of ACAN, COL1A1, RUNX2 and OCN in the 21-day group were increased ( P<0.05 or 0.01), with no significant difference in the expression level of COL2A1 ( P>0.05), but the expression level of SOX9 was decreased ( P<0.05). The contents of collagen in 0-, 7-, 14-and 21-day induction groups were (219.15±0.48)μg/mg, (264.07±1.58)μg/mg, (270.83±0.84)μg/mg and (280.01±0.48)μg/mg respectively. The GAGs contents were (171.18±1.09)μg/mg, (184.06±1.37)μg/mg, (241.08±0.84)μg/mg and (201.14±0.17)μg/mg respectively. Compared with the 0-day induction group, the contents of collagen and GAGs in 7-, 14-, and 21-day induction groups were significantly increased ( P<0.01), among which the content of collagen was the lowest in 7-day induction group ( P<0.01) but the highest in the 21-day induced group ( P<0.01); the content of GAGs was the lowest in the 7-day induced group ( P<0.01) but the highest in the 14-day induction group ( P<0.01). Conclusions:The microcarriers prepared by combining physical and chemical methods are decellularized successfully, with more matrix retention, uniform composition and on cytotoxicity. By loading microcarriers with hUCMSCs and hCho, cartilage organoids are successfully constructed in vitro, which are characterized by good cell activity, self-renewal ability, strong expression of genes related to chondrogenesis and secretion of collagen and GAGs. The cartilage organoids constructed at 14 days of induction have the best chondrogenic activity.

BACKGROUND:Compared with traditional two-dimensional culture,three-dimensional microtissue culture can show greater advantages.However,more favorable cultivation methods in three-dimensional culture still need to be further explored. OBJECTIVE:To evaluate the cell behavior of microtissue and its ability to promote cartilage formation under two three-dimensional culture methods. METHODS:Cartilage-derived microcarriers were prepared by chemical decellularization and tissue crushing.DNA quantification and nuclear staining were used to verify the success of decellularization,and histological staining was used to observe the matrix retention before and after decellularization.The microcarriers were characterized by scanning electron microscopy and CCK-8 assay.Cartilage-derived microtissues were constructed by combining cartilage-derived microcarriers with human adipose mesenchymal stem cells through three-dimensional static culture and three-dimensional dynamic culture methods.The cell viability and chondrogenic ability of the two groups of microtissues were detected by scanning electron microscopy,live and dead staining,and RT-qPCR. RESULTS AND CONCLUSION:(1)Cartilage-derived microcarriers were successfully prepared.Compared with before decellularization,the DNA content significantly decreased after decellularization(P<0.001).Scanning electron microscope observation showed that the surface of the microcarrier was surrounded by collagen,maintaining the characteristics of the natural extracellular matrix of cartilage cells.CCK-8 assay indicated that microcarriers had no cytotoxicity and could promote cell proliferation.(2)Scanning electron microscopy and live and dead staining results showed that compared with the three-dimensional static group,the three-dimensional dynamic group had a more extended morphology of microtissue cells,and extensive connections between cells and cells,between cells and matrix,and between matrix.(3)The results of RT-qPCR showed that the expressions of SOX9,proteoglycan,and type Ⅱ collagen in microtissues of both groups were increased at 7 or 14 days.The relative expression levels of each gene in the three-dimensional dynamic group were significantly higher than those in the three-dimensional static group at 14 days(P<0.05).At 21 days,the three-dimensional static group had significantly higher gene expression compared with the three-diomensional dynamic group(P<0.001).(4)The results showed that compared with three-dimensional static culture microtissue,three-dimensional dynamic culture microtissue could achieve higher expression of chondrogen-related genes in a shorter time,showing better cell viability and chondrogenic ability.

MiRNAs are a class of small non-coding RNAs,which regulate gene expression post-transcriptionally by partial complementary base pairing.Aberrant miRNA expressions have been reported in tumor tissues and peripheral blood of cancer patients.In recent years,artificial intelligence algorithms such as machine learning and deep learning have been widely used in bioinformatic research.Compared to traditional bioinformatic tools,miRNA target prediction tools based on artificial intelligence algorithms have higher accuracy,and can successfully predict subcellular localization and redistribution of miRNAs to deepen our understanding.Additionally,the construction of clinical models based on artificial intelligence algorithms could significantly improve the mining efficiency of miRNA used as biomarkers.In this article,we summarize recent development of bioinformatic miRNA tools based on artificial intelligence algorithms,focusing on the potential of machine learning and deep learning in cancer-related miRNA research.

Objective:To compare the accuracy of dynamic navigation system,fully-guided and Pilot-drill guided implant surgery in anterior dental aesthetic area.Methods:45 patients underwent single tooth implantation in anterior dental aesthetic area were random-ly divided into 3 groups(n=15).Dynamic navigation system imlanting(A),3D printing fully-guided template with ASTRA EV assis-ted orthopedic implantation(B)and Pilot-drill guided implantation(C)were respectively used for the patients in group A,B and C.The deviation between the preoperative design and actual results were measured by CBCT and/or 3D reconstruction with navigation de-sign software or 3SHAPE software.SPSS 25.0 software package was used and(x)±s was used for descriptive analysis,results were pair-wise compared among the 3 groups using ANOVA.Results:Data were normally distributed and the variance of the data was homoge-neous.In group A,the distance deviation of implant top center was(0.571±0.196)mm,the horizontal deviation was(0.405±0.222)mm,the distance deviation of implant apical center was(0.449±0.267)mm,the vertical deviation was(0.312±0.223)mm,the de-viation of the connection angle between the two centers was 2.257°±0.989°.In group B,those were(0.520±0.242)mm,(0.219±0.201)mm,(0.643±0.284)mm,(0.464±0.292)mm and 1.272°±0.951° respectively.In group C those were(1.179±0.365)mm,(0.667±0.276)mm,(1.518±0.566)mm,(0.967±0.444)mm and 2.568°±0.632°.The deviation of all the measurments of group A and B was lower than those of group C(P＜0.05).The deviation of the connection angle between the 2 centers of group B was less than that of group A(P＜0.05),but no statistical significance(P＞0.05)was found in other 4 measured items.Conslusion:The accuracy of the dynamic navigation system and the fully-guided implantation is significantly higher than that of the Pilot-drill guided implantation,but is not statistically significant between dynamic navigation and the fully-guided system.

Objective:To investigate the effects of Sennoside A (SA) on the proliferation, migration, invasion, glycolysis and other malignant biological behaviors of gallbladder carcinoma cells, and to analyze the related mechanisms.Methods:Human gallbladder carcinoma cell lines, NOZ and SGC-996, were cultured in vitro and divided into control group, SA low dose group (L-SA, 25 μmol/L), SA medium dose group (M-SA, 50 μmol/L) and SA high dose group (H-SA, 100 μmol/L), and H-SA+ phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathway activator 740Y-P group, respectively. The proliferation, migration, invasion, apoptosis and glycolysis of gallbladder cancer cells in each group were detected by cell counting assay, Transwell, flow cytometry and glycolysis kit. The protein levels of PI3K, p-PI3K, AKT and p-AKT were detected by Western blot assay. NOZ cells were used to construct tumor model of nude mice, and the mice were divided into saline treatment group and 10 mg/kg SA treatment group. The tumor formation ability of the two groups of mice was compared, and the expression level of Ki-67 in tumor of the two groups was detected by immunohistochemical assay.Results:Compared with control group, the proliferation, migration, invasion, glycolysis ability, the expression levels of p-PI3K and p-AKT were significantly decreased in SA treatment groups, while the apoptosis level was significantly up-regulated, all differences were statistically significant ( P<0.05). Compared with H-SA group, the proliferation, migration, invasion and glycolysis of H-SA+ 740Y-P group cells were up-regulated, while the apoptosis level was significantly decreased, all differences were statistically significant ( P<0.05). In vivo tumorigenesis experiments showed that, compared with the control group, the tumor volume of the SA-treated mice was reduced at day 28 [(1 051.32±130.29) mm 3 vs (575.07±170.54) mm 3, P=0.0003), the tumor weight was reduced [(1.04±0.24) g vs (0.58±0.13) g, P=0.0019], and the average optical density of Ki-67 expression was reduced [(77.00±7.00) vs (33.33±7.51), P=0.0018]. Conclusion:SA can inhibit the proliferation, migration, invasion and glycolysis of gallbladder carcinoma cells by regulating PI3K/AKT pathway.

Objective To systematically review qualitative studies on the experience of virtual reality(VR)rehabilitation in stroke patients,so as to provide references for the clinical practice of stroke rehabilitation.Methods The relevant qualitative studies in PubMed,Web of Science,Scopus,Embase,Cochrane Library,CINAHL,CNKI,WanFang Data,SinoMed and VIP Database were retrieved.The retrieval time was from the establishment of the database to April 2023.The quality of included studies was evaluated according to JBI Critical Appraisal Tool(2020)for qualitative studies.Meta-synthesis was used to integrate results.Finally,the quality of evidence was evaluated by the Confidence in the Qualitative Evidence(ConQual)approach.Results A total of 14 studies were included to extract 45 research results,and 3 synthesized findings were grouped from 10 categories according to their similarities,including the perceived benefits of VR rehabilitation;the perceived barriers of VR rehabilitation;the expectations and needs for VR rehabilitation in stroke patients.The ConQual score showed a moderate quality of the synthesized findings.Conclusion Stroke patients are positive about VR rehabilitation.But in the future,we should pay attention to optimizing the VR rehabilitation support system for stroke,improving the feedback mechanism of VR rehabilitation technology,innovating VR multi-functional rehabilitation form,promoting VR rehabilitation extend to the community and home,providing a reference for the application of VR rehabilitation in stroke patients.