Objective:To explore the effects of different polymers on in vitro biomimetic mineralization of small intestinal submucosa(SIS)scaffolds,and to evaluate the physicochemical properties and bio-compatibility of the SIS scaffolds.Methods:The SIS scaffolds prepared by freeze-drying method were im-mersed in simulated body fluid(SBF),mineralized liquid containing polyacrylic acid(PAA)and mine-ralized liquid containing PAA and polyaspartic acid(PASP).After two weeks in the mineralized solu-tion,the liquid was changed every other day.SBF@SIS,PAA@SIS,PAA/PASP@SIS scaffolds were ob-tained.The SIS scaffolds were used as control group to evaluate their physicochemical properties and bio-compatibility.We observed the bulk morphology of the scaffolds in each group,analyzed the microscopic morphology by environment scanning electron microscopy and determined the porosity and pore size.We also analyzed the surface elements by energy dispersive X-ray spectroscopy(EDX),analyzed the struc-ture of functional groups by Flourier transformed infrared spectroscopy(FTIR),detected the water ab-sorption rate by using specific gravity method,and evaluated the compression strength by universal me-chanical testing machine.The pro-cell proliferation effect of each group of scaffolds were evaluated by CCK-8 cell proliferation method.Results:Under scanning electron microscopy,the scaffolds of each group showed a three-dimensional porous structure with suitable pore size and porosity,and crystal was observed in all the mineralized scaffolds of each group,in which the crystal deposition of PAA/PASP@SIS scaffolds was more regular.At the same time,the collagen fibers could be seen to thicken.EDX analysis showed that the characteristic peaks of Ca and P were found in the three groups of mineralized scaffolds,and the highest peaks were found in the PAA/PASP@SIS scaffolds.FTIR analysis proved that all the three groups of mineralized scaffolds were able to combine hydroxyapatite with SIS.All the scaf-folds had good hydrophilicity.The compressive strength of the mineralized scaffold in the three groups was higher than that in the control group,and the best compressive strength was found in PAA/PASP@SIS scaffold.The scaffolds of all the groups could effectively adsorb proteins,and PAA/PASP@SIS group had the best adsorption capacity.In the CCK-8 cell proliferation experiment,the PAA/PASP@SIS scaffold showed the best ability to promote cell proliferation with the largest number of living cells observed.Con-clusion:Compared with other mineralized scaffolds,PAA/PASP@SIS scaffolds prepared by mineralized solution containing both PAA and PASP have better physicochemical properties and biocompatibility and have potential applications in bone tissue engineering.

OBJECTIVE To investigate the in vitro inhibitory mechanism of berberine on the proliferation of tumor stem cells and evaluate its in vivo safety. METHODS Flow cytometry was used to select tumor stem cells from mouse skin melanoma B16F10 cells； CD44， CD133， Nanog homologous box protein （NANOG） and octamer-binding transcription factor 4 （OCT4） were used as indicators to characterize tumor stem cells. Tumor stem cells were divided into control group， all-trans retinoic acid （ATRA） group， and berberine group， and the CCK-8 method was used to detect the effects of berberine on the viability of tumor stem cells； flow cytometry was adopted to detect cell apoptotic rate， the proportion of CD44+/CD133+ and the positive cell rate of sex determining region Y box protein 2 （SOX2）； the morphological changes of tumor balls were recorded after treatment with berberine； the morphology of cell pyroptosis in each group was recorded， and the release rate of lactate dehydrogenase （LDH） was detected； Western blot assay was adopted to detect the expressions of pyroptosis-related protein gasdermin E （GSDME）， GSDME- N， caspase-3 and cleaved caspase-3. Preliminary evaluation of in vivo safety of berberine was conducted by using zebrafish embryo toxicity experiments. RESULTS Compared with B16F10 cells， the proportion of CD44+/CD133+ cells in tumor stem cells and the fluorescence intensity of NANOG and OCT4 were significantly increased （P＜0.000 1）. The half-inhibitory concentration of berberine to tumor stem cells was 50.98 μmol/L. Compared with the control group， the apoptotic rate of cells in the berberine group was significantly increased， while the proportion of CD44+/CD133+ cells and the rate of SOX2 positive cells were reduced significantly （P＜0.000 1）； tumor stem cell spheroids were atrophied， with partial cell death. After treatment with berberine， tumor stem cells exhibited swelling in their outermost layer， the release rate of LDH of cells was significantly increased and the release rate of LDH increased with increasing dose； the protein expressions of GSDME-N and cleaved-caspase-3 of cells in berberine 20， 40 μmol/L groups were significantly increased， and the protein expressions of GSDME and caspase-3 were significantly reduced （except for berberine 20 μmol/L group， P＜0.05）. The embryonic development of zebrafish treated with berberine was almost unaffected， and the survival rate of embryo reached 100%， with no obvious abnormalities observed. CONCLUSIONS Berberine has good activity against the proliferation of tumor stem cells， and its mechanism of action may be related to activating GSDME and promoting cell pyroptosis； berberine has good in vivo safety.

Objective:To investigate the diagnostic efficacy of targeted biopsy combined with regional systematic biopsy in prostate cancer(PCa)in patients with prostate imaging reporting and data system v2.1(PI-RADS v2.1)4-5.Methods:From January 2023 to October 2023,patients who underwent prostate biopsy for the first time with total prostate specific antigen(tPSA)≤20 ng/mL and had a multi-parametric magnetic resonance imaging(mpMRI)PI-RADS of 4-5 in Peking University First Hospital were prospectively collected.All the patients underwent transrectal ultrasound-guided cognitive fusion tar-geted biopsy(3 cores)followed by systematic biopsy(12 cores).Various hypothetical biopsy schemes were defined based on different biopsy sites.The detection effectiveness of targeted biopsy combined with regional systematic biopsy and other biopsy schemes for prostate cancer were compared using Cochran's Q and McNemar tests.Results:A total of 255 patients were enrolled,of whom 204(80.0％)were de-tected with prostate adenocarcinoma and 187(73.3％)were clinically significant with prostate cancer(csPCa).The detection rate of PCa with targeted biopsy was significantly lower than that of targeted biopsy combined with 12-core system biopsy(77.3％vs.80.0％,P=0.016),and 71.4％(5/7)of the missed patients was csPCa.There was no significant difference in the detection rate between targeted biopsy combined with 4-core regional system biopsy and 12-core system biopsy(P＞0.999),and 1 case of csPCa and clinically insignificant prostate cancer(cisPCa)were missed.There was no significant difference in the detection rate of PCa between targeted combined regional system biopsy and targeted combined lateral or traditional 6-core system biopsy and the number of cores were reduced.Missed diag-nosis of targeted biopsy was correlated with the maximum diameter of the lesion(OR=0.086,95％CI:0.013-0.562,P=0.010).For the patients with PI-RADS 5,only 1 case of PCa was missed in 122 cases by targeted biopsy alone.For patients with PI-RADS 4,6 PCa cases were missed among the 133 patients with targeted biopsy alone,and 1 case of csPCa and cisPCa were missed by targeted biopsy com-bined with regional system biopsy.The statistics of positive core counts for different biopsy schemes indi-cated that targeted combined regional systematic biopsy had a higher proportion of positive cores second only to targeted biopsy alone.Conclusion:Targeted biopsy combined with regional systematic biopsy has high diagnostic efficacy in patients with PI-RADS 4-5 and can be considered as one of the improved schemes for combined biopsy.Targeted biopsy alone is also a feasible option for patients for patients with a PI-RADS score of 5.

BACKGROUND: Glutamine synthetase (GS) and arginase 1 (Arg1) are widely used pathological markers that discriminate hepatocellular carcinoma (HCC) from intrahepatic cholangiocarcinoma; however, their clinical significance in HCC remains unclear. METHODS: We retrospectively analyzed 431 HCC patients: 251 received hepatectomy alone, and the other 180 received sorafenib as adjuvant treatment after hepatectomy. Expression of GS and Arg1 in tumor specimens was evaluated using immunostaining. mRNA sequencing and immunostaining to detect progenitor markers (cytokeratin 19 [CK19] and epithelial cell adhesion molecule [EpCAM]) and mutant TP53 were also conducted. RESULTS: Up to 72.4% (312/431) of HCC tumors were GS positive (GS+). Of the patients receiving hepatectomy alone, GS negative (GS-) patients had significantly better overall survival (OS) and recurrence-free survival (RFS) than GS+ patients; negative expression of Arg1, which is exclusively expressed in GS- hepatocytes in the healthy liver, had a negative effect on prognosis. Of the patients with a high risk of recurrence who received additional sorafenib treatment, GS- patients tended to have better RFS than GS+ patients, regardless of the expression status of Arg1. GS+ HCC tumors exhibit many features of the established proliferation molecular stratification subtype, including poor differentiation, high alpha-fetoprotein levels, increased progenitor tumor cells, TP53 mutation, and upregulation of multiple tumor-related signaling pathways. CONCLUSIONS GS- HCC patients have a better prognosis and are more likely to benefit from sorafenib treatment after hepatectomy. Immunostaining of GS may provide a simple and applicable approach for HCC molecular stratification to predict prognosis and guide targeted therapy.
Humans ; Carcinoma, Hepatocellular/metabolism* ; Sorafenib/therapeutic use* ; Liver Neoplasms/metabolism* ; Glutamate-Ammonia Ligase/metabolism* ; Hepatectomy ; Retrospective Studies ; Prognosis ; Neoplasm Recurrence, Local/surgery*

【Objective】 We combined the concept of traditional medicine with magnetic induction technology, originally brought up the research concept of magnetic hyperthermia to cure KOA, explored the mechanism and constructed a new treatment of KOA with modern medical features. 【Methods】 Through establishing a primary KOA model in rats and constructing ferrimagnetic vortex domain iron oxide nanorings (FVIOs) as a platform for highly efficient magnetic hyperthermia agent, the lesions of KOA were heated accurately under the low-intensity magnetic field. We confirmed the curative effect through the results of pain perception, histopathology, knee joint morphology and microscopic bone structure and the content of serum inflammatory factor, to study the therapeutic mechanism of magnetic hyperthermia for KOA. 【Results】 Compared with the model group, the recovery of mechanical pain threshold after magnetic hyperthermia improved by approximately 48.9%; the degree of hyperemia and edema of joint capsule and synovial tissue and the wear degree of joint cartilage surface, were significantly reduced; the Mankin and OARSI scores decreased by about 33% and 20%, respectively; the MicroCT results indicated that the degree of hardening of the subchondral bone also improved; the expression of inflammatory factors in the serum was reduced. 【Conclusion】 In this study, we utilized the FVIOs as a high-efficiency magnetic hyperthermia platform for the treatment of KOA. The efficacy of magnetic hyperthermia on KOA is clarified, and the mechanism is related to the inhibition of inflammatory factors.

OBJECTIVE: To analyze the clinical phenotype and genetic variant of a child with Snijders Blok-Campeau syndrome (SBCS). METHODS: A child who was diagnosed with SBCS in June 2017 at Henan Children's Hospital was selected as the study subject. Clinical data of the child was collected. Peripheral blood samples of the child and his parents were collected and the extraction of genomic DNA, which was subjected to trio-whole exome sequencing (trio-WES) and genome copy number variation (CNV) analysis. Candidate variant was verified by Sanger sequencing of his pedigree members. RESULTS: The main clinical manifestations of the child have included language delay, intellectual impairment and motor development delay, which were accompanied with facial dysmorphisms (broad forehead, inverted triangular face, sparse eyebrows, widely spaced eyes, narrow palpebral fissures, broad nose bridge, midface hypoplasia, thin upper lip, pointed jaw, low-set ears and posteriorly rotated ears). Trio-WES and Sanger sequencing revealed that the child has harbored a heterozygous splicing variant of the CHD3 gene, namely c.4073-2A>G, for which both of his parents were of wild-type. No pathogenic variant was identified by CNV testing. CONCLUSION The c.4073-2A>G splicing variant of the CHD3 gene probably underlay the SBCS in this patient.
DNA Copy Number Variations ; Heterozygote ; Pedigree ; Phenotype ; RNA Splicing ; Mutation

Objective:To explore the clinical value of modified upper urinary tract video urodynamics in evaluating the surgical effect and guiding the follow-up treatment after ureteral reconstruction.Methods:From December 2018 to November 2020, sixty-nine patients underwent upper urinary tract reconstruction and received modified video urodynamics at the time of nephrostomy removal 3 months after the surgery in the RECUTTER database (29 cases in Peking University First Hospital, 22 cases in Emergency General Hospital, and 18 cases in Beijing Jiangong Hospital). There were 39 males and 30 females, with an average age of (40.4±12.7)years. The stricture was located in left in 34 patients, right side in 27 patients, and bilateral sides in 8 patients. The upper, middle, and lower thirds of the ureter were affected in 26, 10, and 33 cases, respectively. The preoperative creatinine was (92.3±26.9)μmol/L, and the estimated glomerular filtration rate (eGFR) was (85.1±23.2)ml/(min·1.73m 2). The upper urinary tract reconstruction included ileal replacement of ureter in 25 cases (36.2%), pyeloplasty in 8 cases (11.6%), ureteroneocystostomy in 9 cases (13.0%), boari flap in 6 cases (8.7%), lingual mucosal graft ureteroplasty in 9 cases (13.0%), appendiceal onlay ureteroplasty in 3 cases (4.3%), ureteroureterostomy in 3 cases (4.3%), and balloon dilation in 6 cases (8.7%). Based on the pressure and imaging, the results could be divided into three types, type Ⅰ, the pressure difference remained stable near baseline, and the renal pelvis pressure was below 22 cmH 2O(1 cmH 2O=0.098 kPa), and the reconstructed ureter is well visualized during the whole perfusion process; type Ⅱ, the pressure difference increases with the perfusion, but it can decrease to a normal level with the ureteral peristalsis; type Ⅲ, the pressure difference exceeds 15 cmH 2O, and the ureteral peristalsis is weak or disappears at the same time. The management strategies and treatment effects of different subtypes were analyzed. Successful treatment was defined as no further treatment required, the absence of hydronephrosis-related symptoms, and the improved or stabilized degree of hydronephrosis. Results:All 69 patients successfully completed upper urinary tract video urodynamics. The pressure difference was higher than 15 cmH 2O in 8 patients, and the median pressure difference was 37(19-54)cmH 2O. The renal pelvis pressure exceeded 22 cmH 2O in 10 patients, and the median pressure was 63.5 (24-155) cmH 2O. Video urodynamic results of upper urinary tract were classified as type Ⅰ in 60 cases, type Ⅱ in 5 cases, and type Ⅲ in 4 cases. Patients in type Ⅰ do not require other treatment after nephrostomy tube removal. Patients in type Ⅱ should avoid holding urine after the removal of nephrostomy and D-J tubes. All patients in type Ⅲ received further treatment, of which 2 patients replaced D-J tube regularly, 1 patient underwent long-term metal ureteral stent replacement, and 1 patient underwent ureteroscopic balloon dilation. The median follow-up time was 24 (18-42) months. All patients in type Ⅰ met the criteria for surgical success, The pre-and postoperative creatinine in type Ⅰ patients were (88.71±23.09)μmol/L and (88.75±23.64)μmol/L ( P=0.984), and eGFR were (88.06±22.66)ml/(min· 1.73m 2)and (87.97±23.01)ml/(min·1.73m 2), respectively( P=0.969). For type Ⅱ patients, ultrasound showed that the degree of hydronephrosis improved in 3 cases and remained stable in 2 cases. The pre-and postoperative creatinine were (105.97±7.75)μmol/L and (97.63±7.56)μmol/L ( P=0.216), and eGFR were (69.08±14.74)ml/(min·1.73m 2)and (75.95±14.02)ml/(min·1.73m 2)( P=0.243), respectively. For type Ⅲ patients, ultrasound showed that the degree of hydronephrosis remained stable. The pre-and postoperative creatinine were (105.14±44.34)μmol/L and (101.49±57.02)μmol/L ( P=0.684), and eGFR were (65.32±19.85)ml/(min·1.73m 2) and (73.42±27.88) ml/(min·1.73m 2), respectively( P=0.316). Conclusions:The pressure and imaging results of modified upper urinary tract video urodynamics can assist in evaluating the surgical effect of ureteral reconstruction, and the classification has certain guiding significance for further treatment.

BACKGROUND: Anaplastic lymphoma kinase (ALK) rearrangement is a common driver gene of non-small cell lung cancer (NSCLC). Ceritinib is a second-generation ALK inhibitor, which can bring survival benefits to ALK-positive metastatic NSCLC. However, few studies focus on the safety and efficacy of ceritinib in China. Therefore, this study intends to investigate the safety and preliminary efficacy of ceritinib 450 mg with meals in Chinese patients with ALK-positive NSCLC through a real world study. METHODS: From October 2018 to December 2019, patients with ALK-positive NSCLC from 8 medical centers in Sichuan province were recruited in this study. All of these participants received ceritinib 450 mg/d with food. The basic characteristics, adverse effects (AEs) and responses were collected and analyzed in order to evaluate the safety and efficacy of ceritinib. RESULTS: A total of 109 patients were included in this study. Data cutoff was January 23, 2020. The median duration of treatment exposure was 5.87 mon (range: 0.4 mon-15.7 mon). Total AEs were reported in 98 (89.9%) of 109 patients and grade 3 or 4 AEs were reported in 22.9% of patients. Most common AEs (mainly grade 1 or 2) were diarrhea (60.6%), elevated alanine aminotransferase (ALT)(38.5%) and aspartate aminotransferase (AST)(37.6%). As of data cutoff, 45 patients discontinued ceritinib. The overall response rate (ORR) was 37.6% (95%CI: 28.5%-47.4%) and disease control rate (DCR) was 86.2% (95%CI: 78.3%-92.1%). CONCLUSIONS The treatment of ceritinib 450 mg with food for Chinese ALK-positive NSCLC patients had a good safety profile and favorable DCR in real-world setting. However, this conclusion needs to be further verified by large sample, prospective trials.

Objective To investigate the effect ofbiglycan (BGN) on neural apoptosis in mice with early brain injury (EBI) after subarachnoid hemorrhage (SAH).Methods SAH models were induced by endovascular perforation in young male C57BL/6J mice.(1) Totally,48 mice were randomly divided into sham-operated group,SAH 6 h group,SAH 12 h group,SAH 24 h group,SAH 48 h group,and SAH 72 h group (n=8);the BGN protein and mRNA expressions were detected by Western blotting and real-time quantitative PCR (qRT-PCR).(2) Totally,16 mice were randomly divided into sham-operated group and SAH 48 h group (n=8);immunofluorescent double staining was conducted to explore the BGN expression in the neurons of brain tissues.(3) Totally,24 mice were randomly divided into sham-operated group,sham+control lentivirus group,and sham+BGN lentivirus group (n=8);BGN lentiviral vector and control lentivirus were administered intracerebroventricularly 7 d before sham-operation;qRT-PCR was performed to explore the BGN mRNA expression.(4) Totally,48 mice were randomly divided into sham-operated group,SAH+control lentivirus group,and SAH+BGN lentivirus group (n=16);BGN lentiviral vector and control lentivirus were administered intracerebroventricularly 7 d before SAH;neurological scores were detected by modified Garcia scale and beam balance tests;TUNEL was used to detect the neuronal apoptosis,and Western blotting was performed to explore the expressions of nuclear transcription factor kappa B (NF-κB) and phosphorylated-(p-) NF-κB.Results (1) Mice in the SAH 48 h group had the highest BGN protein and mRNA expressions,which showed statistical differences as compared with the sham-operated group (P＜0.05).(2) A majority of BGN expressions were detected in the neurons 48 h after SAH.(3) The sham+BGN lentivirus group had statistically lower BGN mRNA expression than the sham+control lentivirus group (P＜0.05).(4) As compared with those in the SAH+control lentivirus group,both scores of modified Garcia scale and beam balance tests were significantly higher in SAH+BGN lentivirus group (6.125±1.246 vs.13.000±1.309;1.125±1.126 vs.2.875±0.835),and neural apoptosis ratio and ratio of p-NF-κB/NF-κB were significantly lower in the SAH+BGN lentivirus group (51.950％±11.166％ vs.31.938％±7.705％;1.161±0.156 vs.0.886±0.142,P＜0.05).Conclusion Inhibition of BGN can effectively reduce neuronal apoptosis in mice with EBI after SAH,and attenuate neurological deficits.

Objective To investigate the effect of long non-coding RNA F19 (lncRNA F19) on secondary brain injury following traumatic brain injury (TBI) in mice. Methods (1) A total of 96 C57BL/6J male wild-type mice were divided into sham group, sham+control lentivirus group, sham+F19 lentivirus group, TBI group, TBI+control lentivirus group and TBI+F19 lentivirus group according to the random number table. Each group consisted of two subgroups of 1 day and 3 days after TBI, with eight mice per subgroup. The expression and silence efficiency of lncRNA F19 were detected. ( 2 ) A total of 96 C57BL/6J male wild-type mice were divided into sham group, TBI+control lentivirus group and TBI + F19 lentivirus group according to the random number table. Each group consisted of two subgroups of 1 day and 3 days after TBI, with 16 mice per subgroup. The effect of lncRNA F19 on neuronal apoptosis after TBI was recorded. The mice TBI model was established using the controlled cortical damage method (CCI). The lncRNA F19 lentivirus or control lentivirus were administrated by intracerebroventricular injection 5 days before injury. The expressions of lncRNA F19 ( 2 -ΔΔct ) were detected by real-time quantitative PCR ( qRT-PCR ) at 1 day and 3 days after injury. The Toll-like receptor 4 (TLR4), B lymphocyte tumor-2 (Bcl-2) and Bcl-2 related protein (Bax) expressions were detected by Western blot. The TUNEL was used to detect apoptosis around the traumatic lesions. Results From the first day after injury, both in the sham operation and TBI groups, the control lentivirus had no effect on the level of lncRAN F19 (P >0. 05). One day after injury, compared with sham +control lentivirus group, the levels of lncRNA F19 in sham + F19 lentivirus group were significantly decreased (0. 07 ± 0. 07:0. 93 ± 0. 17);compared with TBI+control lentivirus group, levels of lncRNA F19 in TBI+F19 lentivirus group were significantly decreased (2. 91 ± 1. 18:0. 52 ± 0. 32) (P<0. 05). There were significantly lower protein levels of TLR4 (0. 51 ± 0. 13:0. 66 ± 0. 15), Bax (0. 45 ± 0. 06:0. 67 ± 0. 16), lower TUNEL-positive neurons ratio [(23. 55 ± 6. 85)％ : (31. 58 ± 7. 52)％], but higher protein levels of Bcl-2 (0. 76 ± 0. 16:0. 47 ± 0. 12) in TBI+F19 lentivirus group compared with the TBI+ control lentivirus group (P <0.05). Three days after injury, compared with sham + control lentivirus group, levels of lncRNA F19 in sham+F19 lentivirus group were significantly decreased (0. 11 ± 0. 09:0. 96 ± 0. 09); compared with TBI+control lentivirus group, levels of lncRNA F19 in TBI+F19 lentivirus group were significantly decreased (0. 54 ± 0. 24:3. 39 ± 0. 90) (P <0. 05). There were significantly lower protein levels of TLR4 (0. 60 ± 0. 20):(0. 85 ± 0. 09)], lower Bax (0. 60 ± 0. 12:0. 88 ±0. 21), lower TUNEL-positive neurons ratio [(29. 10 ± 7. 37)％ :(39. 22 ± 10. 64)％], but higher protein levels of Bcl-2 (0. 66 ± 0. 12:0. 35 ± 0. 16) in TBI+F19 lentivirus group compared with the TBI+control lentivirus group (P<0. 05). Conclusion Inhibition of lncRNA F19 can significantly reduce the TLR4-induced neuronal apoptosis in cortex after TBI in mice and alleviate reduce the secondary brain injury.