Objective To investigate the effects of nerve growth factor (NGF) on the expression of 67-kDa laminin receptor (67LR) in human bile duct carcinoma QBC939 cells, and study the possible mechanism of perineural invasion and metastasis of bile duct carcinoma. Methods ( 1 ) The expression of a high-affinity receptor for NGF, TrkA, was detected by immunofluorescence staining. ( 2 ) QBC393 cells were pretreated by β-NGF at different concentrations ( 1, 10, 100,200 μg/L), and then the mRNA and protein expressions of 67LR were examined by Real-Time PCR and Western blot assay. QBC939 cells were divided into control group and β-NGF (1, 10, 100,200 μg/L) groups. (3) The ideal concentration of β-NGF was selected according to the results of previous tests, and then the mRNA and protein expressions of 67LR were re-examined by adding specific TrkA inhibitor K252a at different concentrations ( 100,200,300 nmol/L). QBC939 cells were divided into control group, β-NGF 100 μg/L group and K252a ( 100,200,300 nmol/L) groups. All data were analyzed by one-way analysis of variance or LSD-test. Results (1) A strong expression of TrkA was detected in the membrane of QBC939 cells. (2) The mRNA and protein expressions of 67LR in QBC939 cells were 0.35 ± 0.06 and 0. 32 ± 0.05 in the control group, 0.38 ±0.14 and 0.50 ±0.09 in the β-NGF 1 μg/L group, 0.62 ±0.14 and 0. 69 ±0. 13 in β-NGF 10 μg/L group, 0.90 ± 0.08 and 0.93 ± 0.07 in the β-NGF 100 μg/L group, and 0. 70 ± 0. 10 and 0. 76 ±0.07 in the β-NGF 200 μg/L group, there were significant differences among the five groups (F = 22. 4, 14. 6,P ＜0.05). The mRNA and protein expressions of 67LR in the β-NGF 100 μg/L group were significantly higher than those in the control group ( t = 19. 0, 21.0, P ＜ 0. 05 ). (3) The mRNA and protein expressions of 67LR in the QBC939 cells were 0.35 ±0.10 and 0.41 ±0.10 in the control group, 0. 88 ±0. 14 and 0.84 ±0.10 in the β-NGF 100 μg/L group, 0.80±0.08 and 0.76 ±0.04 in the K252a 100 nmol/L group, 0.67 ±0.12 and 0.61 ± 0.09 in the K252a 200 nmol/L group, and 0. 43 ± 0.07 and 0. 50 ± 0. 12 in the K252a 300 nmol/L group, there were significant differences among the five groups ( F = 14. 1, 8. 9, P ＜ 0.05 ). There were no significant differences in the mRNA and protein expressions of 67LR between the K252a 300 nmol/L group and the control group (t =1.02, 0. 85, P＞0.05). Conclusion In bile duct carcinoma cells, NGF enhances the expression of 67LR by combining with TrkA, which might be the mechanism of NGF mediating perineural invasion of bile duct carcinoma.

Objective To investigate the correlation between α1-adrenergic receptor and the pathological behavior of cholangiocarcinoma, and the effects of norepinephrine (NE) on the proliferation of cholangiocarcinoma cell line QBC939. Methods Thirty-six samples of cholangiocarcinoma were resected in Southwest Hospital from August 2002 to March 2008. The expression of α1-adrenergic receptor in the 36 samples of cholangiocarcinoma tissue and 4 samples of normal bile duct tissue were detected by SABC technique. The proliferation of cholangio-carcinoma cell line QBC939 was detected after processing the cells with NE, phentolamine and prazosin. All the data were analyzed by chi-square test. Results The high positive expression rate of α1-adrenergic receptor was 68% (17/25) in patients with lymph node metastasis, which was significantly higher than 9% (1/11) in patients without lymph node metastasis (χ2=10.604, P<0.05). The high positive expression rate of α1-adrenergic receptor was 85% (11/13) in patients with middle and low positioned cholangiocarcinoma, which was significantly higher than 30% (7/23) in patients with hilar cholangiocarcinoma (χ2=9.753, P<0.05). NE promoted the proliferation of cholangiocarcinoma cell line QBC939 by stimulating the expression of α1-adrenergic receptor, and in a concentration-dependent manner. The proliferative effect was weakened as time passed by, and it was eliminated by phentolamine and prazosin. Conclusions The expression of α1-adrenergic receptor is diverse due to lymph node metastasis and the location of the tumor, α1-adrenergic receptor with high expression may play an important role in the proliferation and metastasis of cholangiocarcinoma.

Objective To investigate the clinical significance of multiple tumor markers in the early diagnosis of cholangiocarcinoma.Methods 165 patients with histopathologically confirmed cholangiocarcinoma and 25 patients with gallstone canceration were enrolled in this study.36 patients with gallstones and 46 patients with bile duct stenosis were enrolled as controls.The determinations of multiple tumor markers,including carcinoembryonic antigen (CEA),CA125,CA153,CA242,CA19-9,alpha-fIetoprotein (AFP) and Ferritin,of all subjects and the imageological determinations of all the patients with cholangiocarcinoma and with gallstone canceration were analysed retrospectively.Comparisons between the groups were performed using the likelihood ratio test of the Chi-square test.Results CA19-9 had the highest sensitivity and specificity (83.6％ and 96.3％ respectively) for cholangiocarcinoma,and CA242 had a high sensitivity for gallstone canceration though it was not so sensitive in cholangiocarcinoma.Multiple tumor markers increased the sensitivity and specificity in the early diagnosis of cholangiocarcinoma.Compared with imageological determinations,multiple tumor markers had a higher sensitivity for the early diagnosis of cholangiocarcinoma with a tumor mass ＜1 cm.Conclusion Multiple tumor markers have important clinical values in the early diagnosis of cholangiocarcinoma.

Objective To assess the local treatment efficacy of percutaneous radiofrequency ablation(PRFA) in combination with percutaneous ethanol injection(PEI) therapy protocol for liver metastases in patients with colorectal cancer.Methods From February 2000 to February 2004, total of 61 liver metastatic lesions(1.5～7.8 cm) from 37 colorectal cancer patients were ablated by using RF-2000 generator and 10-needle LeVeen electrode percutaneously guided by B-ultrasound.Tumors larger than 2.5 cm in diameter were treated by multiple overlapping radiofrequency ablations encompassing the entire lesion as well as a rim of normal liver tissue(approximately 1.0 cm).PEI was performed additionally.The Chiba needle and Quadra-Fuse multi-pronged injection needle were used for tumors ranged 1～2 cm and for those larger than 3 cm respectively.Results The procedures were performed successfully in all patients.The post-PRFA/PEI complication rate was 5.4%.The follow-up rate was 97.3%(36/37).Thirteen patients achieved a complete response(CR),and 21 achieved a partial response(PR).The effective rate(CR+PR) was 94.4%.The patients' survival rates were 97.2% at 1 year,75.0% at 2 years,and 52.8% at 3 years respectively.Conclusion The results of this study suggest that the treatment protocol of PRFA in combination with PEI was an effective mini-invasive local ablation therapy,which was part of the comprehensive management of the colorectal liver metastases.

Diabetic retinopathy(DR)represents the primary cause of blindness among the global working-age population, and the disruption of the blood-retinal barrier is a crucial factor. Research in recent years has elucidated that DR transcends the scope of a mere microvascular disorder into a complex interplay of retinal glial cells and neurodegeneration microvascular pathology. Neuronal damage may precede vascular endothelial changes in the retinal neurovascular unit(RNVU)in the early stage of DR, and glial cell activation further exacerbates vascular barrier dysfunction. Retinal microglia are immune cells that reside in the retina and are involved in chronic inflammatory responses induced by long-term exposure to high glucose levels. Microglia secrete various inflammatory factors in response to high glucose levels, which can lead to the destruction of the blood-retinal barrier structure, increased neuronal apoptosis, and altered gliosis of Muller cells, thus affecting the retina's homeostatic balance. The RNVU has received increasing attention in recent years as a unitary structural study, and the mechanism of microglia in the RNVU and the progress of the study are reviewed.

Objective To analyze the clinicopathological features and prognosis of polyomavirus nephropathy (PyVN) after kidney transplantation. Methods Clinical data of 44 patients who were diagnosed with PyVN after kidney transplantation were retrospectively analyzed. The causes of puncture and the time of pathological diagnosis were analyzed. Histological grading was carried out according to Banff 2018 classification. Clinical data and pathological characteristics of patients at all grades were statistically compared. BK viral DNA loads in the blood and urine were measured and renal allograft function were assessed. Clinical prognosis of all patients was compared among different groups and the risk factors affecting clinical prognosis were also analyzed. Results The time interval between pathological diagnosis of PyVN and kidney transplantation was 16(8, 29) months, and the increase of serum creatinine level was the main cause for puncture. Among 44 patients, 19 cases were classified as grade ⅠPyVN, 21 cases of grade Ⅱ PyVN and 4 cases of grade Ⅲ PyVN, respectively. Under optical microscope, there was no significant difference in the positive rate of virus inclusion bodies among different groups (P=0.148). Inflammatory cell infiltration, interstitial fibrosis and polyomavirus load in grade Ⅱ PyVN patients were all more or higher than those in grade Ⅰ counterparts. Inflammatory cell infiltration and polyomavirus load in grade Ⅲ patients were more or higher than those in grade Ⅰ counterparts. Polyomavirus load in grade Ⅲ patients was more or higher than that in grade Ⅱ counterparts. The differences were statistically significant (all P < 0.05/3). Upon diagnosis, BK viral DNA load was detected in the blood and urine of 39 patients. Among them, 38 patients were positive for BK virus in the urine and 30 patients were positive for BK virus in the blood. The serum creatinine level upon diagnosis was higher compared with that at postoperative 1 month. The serum creatinine level at the final follow-up was significantly higher than that upon diagnosis. The differences were statistically significant (P < 0.001, P=0.049). There was no significant difference in the serum creatinine level among patients with different grades of PyVN at postoperative 1 month (P=0.554). The serum creatinine level of patients with grade Ⅱ PyVN upon diagnosis was significantly higher than that of those with grade Ⅰ PyVN (P=0.007). The 1-, 3- and 5-year cumulative survival rates of renal allografts were 95%, 69% and 62%, respectively. The survival rates of renal allografts significantly differed among patients with different grades of PyVN. The higher the grade, the lower the survival rate (P=0.014). Univariate and multivariate Cox's regression analyses prompted that intrarenal polyomavirus load and serum creatinine level upon diagnosis were the independent risk factors for renal allograft dysfunction (all P < 0.05). Conclusions PyVN mainly occurs within 2 years after kidney transplantation. Clinical manifestations mainly consist of increased serum creatinine level, BK viremia and BK viruria. Postoperative routine monitoring of BK virus contributes to early diagnosis and protection of renal allografts. Banff 2018 classification may effectively predict the prognosis of renal allografts.

BACKGROUND:Due to the mechanical properties,unstable drug release,single function and other problems of pure hydrogel materials,in recent years,researchers have prepared a variety of metal organic frameworks-based hydrogel materials by introducing metal organic frameworks into hydrogel,and showed great potential in the field of soft and hard tissue regeneration. OBJECTIVE:To classify the metal organic frameworks-based hydrogel materials based on how metal organic frameworks enhance the properties of hydrogel and further summarize its recent research in the field of soft and hard tissue regeneration,in order to provide ideas and theoretical supports for the subsequent in-depth research on synthesis mechanism and clinical application of the composite material. METHODS:Using"metal organic frameworks,hydrogels,tissue engineering,tissue,bone regeneration,bone,wound"as English and Chinese search terms,we searched Web of Science,PubMed,CNKI,and Wanfang databases.The search period ranged from January 2000 to August 2023.By reading the titles and abstracts,the repetitive studies and unrelated literature of Chinese and English literature were excluded.After the literature quality evaluation,73 articles were included for review. RESULTS AND CONCLUSION:(1)Metal organic frameworks-based hydrogel materials effectively solve the problems of poor mechanical properties,unstable drug release and single function of pure hydrogel.(2)Metal organic frameworks enhance the capacity of repair and regeneration by strengthening the cross-linking of hydrogel,the drug delivery capacity of hydrogel and the multifunction of hydrogel.(3)In terms of hard tissue repair,it has shown good repair effects in animal models of diseases such as bone defects,osteoarthritis,and cartilage defects,suggesting potential application prospects in clinical repair.(4)In terms of soft tissue regeneration,it has the capacities of hemostasis,antibacterial,inflammatory state regulation,oxidative stress state regulation,promoting angiogenesis and other functions,effectively improving the microenvironment of various complex wounds and promoting soft tissue regeneration.(5)Although metal organic frameworks-based hydrogels have many excellent properties,they are still in the initial stage and there are some urgent problems to be solved in the process of clinical transformation,such as the cytotoxicity of metal organic frameworks and large-scale synthesis of metal organic frameworks.(6)With further research,metal organic frameworks-based hydrogels have broad application prospects in the field of soft and hard tissue repair.

This review summarizes the clinical data of one pediatric liver transplant recipient and two adult kidney transplant recipients with posterior reversible encephalopathy syndrome(PRES)at Tongji Hospital of Huazhong University of Science & Technology.The relevant clinical characteristics of recipients are discussed for providing reference for clinical diagnoses and treatments.