Objective To explore the protective effect of Lycium barbanun glycopeptide(LbGP)against osteogenic inhibition induced by exosomes derived from human gingival fibroblasts(HGFs)exposed to radiation.Methods Cultured HGFs with or without LbGP pretreatment were exposed to 8 Gy X-ray radiation,and the changes in cell apoptosis,senescence and α-SMA level were detected using RT-qPCR,Western blotting and β-galactosidase staining.The exosomes secreted by the treated cells were extracted,and after identification by electron microscopy,particle size analysis and Western blotting,the exosomes were added into primary cultured bone mesenchymal stem cells(BMSCs),and osteoclast activity and osteogenesis in the cell cultures were detected by Trap staining,Alizarin red staining,ALP staining,RT-qPCR and Western blotting.Results In cultured HGFs,X-ray radiation significantly increased the percentage of senescent cells,which was obviously lowered by LbGP treatment.X-ray radiation significantly reduced Bcl-2/Bax ratio and increased α-SMA expression in HGFs,and these changes were significantly suppressed by LbGP pretreatment.In rat BMSCs,incubation with the exosomes derived from HGFs with radiation exposure caused a significant increase of osteoclasts,reduced calcium nodules and lowered alkaline phosphatase expression in the cells;The opposite changes were observed in the cells treated with exosomes from LbGP-pretreated HGFs,which also significantly increased the cellular expressions of the osteogenic genes(BMP2,ALP,and RUNX2)and proteins(ALP and RUNX2)as compared with the exosomes from irradiated HGFs.Conclusion LbGP can effectively inhibit osteoclast activity and promote osteogenesis by acting on exosomes secreted by irradiated HGFs,suggesting its potential value for treatment of osteoradionecrosis of the jaw.

Objective:To explore the related factors of vascular complications after liver transplantation in children.Methods:This is a retrospective case series research. The clinical data of 89 pediatric liver transplant patients admitted to the Organ Transplantation Center, the Affiliated Hospital of Qingdao University from January 2016 to March 2024 were collected retrospectively. This study included 44 males and 45 females,aged from 4 months to 17 years. The ratio of graft to recipient weight was 0.6% to 7.7%. The primary diseases included 48 cases of biliary atresia and 41 cases of non-biliary atresia. The Wilcoxon rank sum test, χ2 test, and Fisher′s exact probability method were used for data analysis. Multivariate Logistic regression was used to analyze the related factors of vascular complications. Results:All 89 children with liver transplantation completed surgery successfully. There were 8 cases of arterial complications after surgery, including 6 cases of hepatic artery thrombosis and 2 cases of hepatic artery stenosis. There were 16 cases of portal vein complications after surgery, including 9 cases of portal vein stenosis and 7 cases of portal vein thrombosis. The results of univariate analysis showed that the age of the recipient ≤1 year was the relevant factor for hepatic arterial complications( χ 2=4.772, P=0.029). The age of the recipient ≤1 year, the age of the donor, the hepatic phase, and the time of cold ischemia were the relevant factors for the occurrence of portal vein complications( χ 2=7.270, Z=388.500, Z=838.000, Z=894.500;all P<0.05). The results of multivariate analysis showed that age(≤1 year vs. >1 year) and duration of cold ischemia(every additional 1 hour) were independent related factors for portal vein complications after liver transplantation in children(both P<0.05). Conclusion:Children aged ≤1 year and with prolonged cold ischemia are more likely to develop portal vein complications after liver transplantation.

Small cell lung cancer(SCLC)is a type of lung cancer with high malignant degree,rapid transformation,rapid invasion and metastasis,which is prone to early metastasis and poor prognosis.Bone metastases of SCLC occur in three stages:cancer cells proliferate at the primary site,break through local tissues,enter the blood circulation to form circulating tumor cells(CTCs),reach bone tissue through blood circulation,and take root and germinate to form new tumor sites with the support of the bone microenvironment.However,traditional imaging and pathology examinations have disadvantages such as low sensitivity,high cost and difficulty in implementation.Exploratory studies based on blood marker detection as screening and efficacy evaluation of SCLC bone metastases have been reported in recent years.By reviewing the molecular biological mechanism of SCLC bone metastasis formation,this paper found that conventional diagnostic methods such as imaging and pathological biopsy were inadequate in SCLC bone metastasis.The changes in hyaluronic acid,protein biomarkers,non-coding RNA,and biomarkers in liquid biopsy were earlier than the changes in imaging,which had the advantages of simple operation and good repeatability.It provides a new idea and method for the early diagnosis of SCLC bone metastasis,which is worthy of clinical application.

Objective:To explore the related factors of vascular complications after liver transplantation in children.Methods:This is a retrospective case series research. The clinical data of 89 pediatric liver transplant patients admitted to the Organ Transplantation Center, the Affiliated Hospital of Qingdao University from January 2016 to March 2024 were collected retrospectively. This study included 44 males and 45 females,aged from 4 months to 17 years. The ratio of graft to recipient weight was 0.6% to 7.7%. The primary diseases included 48 cases of biliary atresia and 41 cases of non-biliary atresia. The Wilcoxon rank sum test, χ2 test, and Fisher′s exact probability method were used for data analysis. Multivariate Logistic regression was used to analyze the related factors of vascular complications. Results:All 89 children with liver transplantation completed surgery successfully. There were 8 cases of arterial complications after surgery, including 6 cases of hepatic artery thrombosis and 2 cases of hepatic artery stenosis. There were 16 cases of portal vein complications after surgery, including 9 cases of portal vein stenosis and 7 cases of portal vein thrombosis. The results of univariate analysis showed that the age of the recipient ≤1 year was the relevant factor for hepatic arterial complications( χ 2=4.772, P=0.029). The age of the recipient ≤1 year, the age of the donor, the hepatic phase, and the time of cold ischemia were the relevant factors for the occurrence of portal vein complications( χ 2=7.270, Z=388.500, Z=838.000, Z=894.500;all P<0.05). The results of multivariate analysis showed that age(≤1 year vs. >1 year) and duration of cold ischemia(every additional 1 hour) were independent related factors for portal vein complications after liver transplantation in children(both P<0.05). Conclusion:Children aged ≤1 year and with prolonged cold ischemia are more likely to develop portal vein complications after liver transplantation.

Objective To explore the protective effect of Lycium barbanun glycopeptide(LbGP)against osteogenic inhibition induced by exosomes derived from human gingival fibroblasts(HGFs)exposed to radiation.Methods Cultured HGFs with or without LbGP pretreatment were exposed to 8 Gy X-ray radiation,and the changes in cell apoptosis,senescence and α-SMA level were detected using RT-qPCR,Western blotting and β-galactosidase staining.The exosomes secreted by the treated cells were extracted,and after identification by electron microscopy,particle size analysis and Western blotting,the exosomes were added into primary cultured bone mesenchymal stem cells(BMSCs),and osteoclast activity and osteogenesis in the cell cultures were detected by Trap staining,Alizarin red staining,ALP staining,RT-qPCR and Western blotting.Results In cultured HGFs,X-ray radiation significantly increased the percentage of senescent cells,which was obviously lowered by LbGP treatment.X-ray radiation significantly reduced Bcl-2/Bax ratio and increased α-SMA expression in HGFs,and these changes were significantly suppressed by LbGP pretreatment.In rat BMSCs,incubation with the exosomes derived from HGFs with radiation exposure caused a significant increase of osteoclasts,reduced calcium nodules and lowered alkaline phosphatase expression in the cells;The opposite changes were observed in the cells treated with exosomes from LbGP-pretreated HGFs,which also significantly increased the cellular expressions of the osteogenic genes(BMP2,ALP,and RUNX2)and proteins(ALP and RUNX2)as compared with the exosomes from irradiated HGFs.Conclusion LbGP can effectively inhibit osteoclast activity and promote osteogenesis by acting on exosomes secreted by irradiated HGFs,suggesting its potential value for treatment of osteoradionecrosis of the jaw.

Robotics ; Robotic Surgical Procedures ; Kidney Transplantation

Objective To explore the diagnostic value of radial ultrasound (EBUS) combined with virtual navigation (VBN) in peripheral pulmonary nodules.Methods Two hundreds and forty cases of peripheral pulmonary nodules(0.8 cm≤diameter≤3 cm) in the respiratory department of our hospital from July 2014 to July 2015 were included and according to the different guide de vices,which were divided into the radial ultrasound combined with virtual navigation group (EBUS+ VBN group),radial ultrasound group (EBUS group),virtual navigation group (VBN group) and control group.The diagnostic rates were compared among the four groups and among different sizes of lesion.The time of lesion location and operating time were also compared between the EBUS+ VBN group and EBUS group.Results Among 240 cases,the diagnostic rate in the EBUS+VBN group was highest(81.67％),and the diagnostic rates had statistically significant difference among the four groups (x2=19.344,P=0.00);the diagnostic rates of lesions less than 2 cm in diameter were lower than that of lesions ＞2 cm in the EBUS+-VBN group and EBUS group,but without statistically significant difference (x2 =2.04,3.40,P =0.15,0.07);the locating lesions time and operating times in the EBUS+ VBN group were shorter than those in the EBUS group,but the difference between them was not statistically significant (P=0.03,0.04).Conclusion EBUS combined with VBN could improve the diagnostic rate of peripheral pulmonary nodules and shorten the time of lesion location and operating time.

Objective To investigate the influence of S100A6 gene RNA interference on the biological behaviors of A549 lung adenocarcinoma cells.Methods The S100A6 gene RNA interference vector was transfected in A549 lung adenocarcinoma by lentivirus.The experiment was divided into three groups:pLenR-GPH group (the vector without S100A6 RNAi gene was transfected),negative control group (no vectors was transfected),and RNAi group (the vector with S100A6 RNAi gene was transfected).S100A6 mRNA and protein were detected using real-time PCR and Western blotting.The biological behavior including cell proliferation,invasion,cell cycle and cell apoptosis were detected by 3-(4,5-dimethyl-2-thiazoly)-2,5-diphenyl-2H-tetrazolium bromide,transwell,and flow cytometer,respectively.Results The expression of S100A6 mRNA of A549 lung adenocarcinoma cell line in RNAi group (0.009 ± 0.001) was significantly decreased than those in negative control group (0.049 ± 0.005) and pLenR-GPH group (0.030 ± 0.006),with statistically significant differences (t =57.56,P =0.000;t =48.21,P =0.000).The expression of S100A6 protein of A549 lung adenocarcinoma cell line in RNAi group (0.107 ± 0.002) was significantly decreased than those in negative control group (0.341 ± 0.005) and pLenR-GPH group (0.311 ± 0.006),with statistically significant differences (t =37.34,P =0.000;t =27.51,P =0.001).The ability of cell proliferation at 48 hours in RNAi group (0.230 ± 0.008) was significantly declined than those in negative control group (0.292 ± 0.038) and pLenR-GPH group (0.307 ± 0.013),with statistically significant differences (t =25.31,P =0.003;t =29.42,P =0.001).The number of transmembrane cells in RNAi group (11.40 ± 1.36) was significantly declined than those in negative control group (26.80 ± 1.83) and pLenR-GPH group (25.80 ± 1.93),with statistically significant differences (t =29.44,P =0.001;t =23.17,P =0.005).The cell proportion of S phase in RNAi group (28.26％ ± 0.38％) was significantly lower than those in pLenR-GPH group (44.73％±0.66％) and negative control group (45.15％ ± 1.69％),with statistically significant (t =63.69,P=0.000;t =71.55,P =0.000).Cell propotion of G2-M phase in RNAi group (26.99％ ± 0.29％) was signi-ficantly higher than those in negative control group (13.26％ ±0.49％) and pLenR-GPH group (12.41％ ± 0.46％),with statistically significant (t =56.31,P =0.000;t =51.39,P =0.000).The cell apoptosis proportion in RNAi group (8.90％ ±0.48％) was significantly higher than those in negative control group (5.84％ ±0.21％) and pLenR-GPH group (5.99％ ±0.37％),with statistically significant (t=51.34,P =0.000;t =47.27,P =0.000).Conclusion S100A6 gene involves the proliferation,invasion,cell cycle and apoptosis of tumor cells,which has close correlation with occurrence,development and metastasis of lung adenocarcinoma.S100A6 gene is hopeful to become a molecular target for the diagnosis and treatment of lung adenocarcinoma.

Objective To explore the learning and memory impairment and pathology in hippocampus in rats after spinal cord contu-sion. Methods 36 adult female Sprague-Dawley rats were randomly divided into sham group (n=18) and spinal cord injury group (n=18). Spinal cord injury model at T10 was established with modified Allen's technique (10 g × 25 mm). The hindlimbs behavior of rats was rated with Basso-Beattie-Bresnahan (BBB) scores once a week for 5 weeks. They were tested with motor evoked potentials (MEP) and Morris wa-ter maze 5 weeks after injury. The pathology of hippocampus was detected with HE staining 1 week, 3 weeks and 5 weeks after injury, 4 rats in a group, repectively. Results The BBB scores were significantly lower in the spinal cord injury group than in the sham group at each time point (P<0.05). The latencies of both N1 and P1 wave of MEP were significantly longer in the spinal cord injury group than in the sham group (P<0.001), while the amplitudes were significantly less (P<0.001). For the Morris water maze, the latency of arrival platform were sig-nificantly longer in the spinal cord injury group than in the sham group (P<0.001), and the time in target was significantly less (P<0.001), with more systematic positioning or annular positioning, while the sham group with more space-based positioning. Morphologically abnor-mal cells in hippocampus gradually increased since the first week after injury, with the decrease of cells survival, while it was normal in the sham group. Conclusion Spinal cord contusion can cause learning and memory impairment in rats, which may be related to injury in hippo-campus.

Objective To study the differentiation and proliferation ability of the spinal neural stem cells (NSCs) at different gestational ages in fetal rats. Methods Sprague-Dawley fetal rats were divided into group A (12 days of pregnancy), group B (14 days of pregnancy) and group C (16 days of pregnancy). NSCs were separated with enzyme-assisted microdissection. The diameter and numbers of NSCs balls were measured at different time. The cell growth curve was drawn with CCK8 colorimeter. NSCs were identified with BrdU/Nestin immunohistochemical staining. They were induced with 10% fetal bovine serum for 10 days, and the expression of β-tubulinⅢ and glial fibrillary acidic protein was detected with immunocytochemistry. Results There were cells expressed BrdU, Nestin, β-tubulinⅢ and GFAP in all the group. The most cells (22.74±0.79%) expressed β-tubulinⅢ in the group B, but no significant difference between group B and group C. The cell vitality on the 5th day of third-generation neural stem cells was the most in group B. Conclusion For enzyme-assisted microdissection, it may obtain more neurons to isolate the neural stem cells from 14 days of pregnancy pregnant rats.