Protective Mechanism of Paeoniflorin on Mice with Ulcerative Colitis Based on AMPK/mTOR Autophagy Pathway / 中国实验方剂学杂志

ObjectiveTo explore the protective mechanism of paeoniflorin on mice with ulcerative colitis （UC） through the adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR） autophagy pathway. MethodUC mouse model was established by allowing mice freely drink 4% DSS， and 56 BALB/c male mice were randomly divided into model group， AMPK inhibitor group （20 mg·kg-1）， paeoniflorin （50 mg·kg-1） + inhibitor （20 mg·kg-1） group， and high dose （50 mg·kg-1）， medium dose （25 mg·kg-1）， and low dose （12.5 mg·kg-1） paeoniflorin groups. After seven days of drug intervention， the protective effect of paeoniflorin on mice with UC was determined by comparing the body weight， disease activity index （DAI） changes， and Hematoxylin-eosin （HE） staining results. Enzyme linked immunosorbent assay （ELISA） was used to detect the levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in the serum of mice in each group， and immunofluorescence was utilized to detect microtubule-associated protein 1 light chain 3 （LC3） content in the colon， AMPK， mTOR proteins， and their phosphorylated proteins including p-AMPK and p-mTOR in the colon tissue were detected by Western blot， and the mRNA expression levels of AMPK， mTOR， Beclin1， LC3， and p62 were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. ResultCompared with the blank group， the model group showed a decrease in body mass， an increase in DAI score， and severe pathological damage to the colon. The levels of inflammatory factors including TNF-α and IL-6 increased in serum （P<0.01）， while the protein levels of LC3 and p-AMPK/AMPK were down-regulated in colon tissue， and those of p-mTOR/mTOR were up-regulated （P<0.01）. The mRNA expression levels of AMPK and LC3 were down-regulated， while the mRNA expression levels of mTOR and p62 were up-regulated （P<0.01）. Compared with the model group and the paeoniflorin + inhibitor group， the mice treated with paeoniflorin showed an increase in body mass， a decrease in DAI score， a reduction in pathological damage to colon tissue， and a reduction in the levels of inflammatory factors of TNF-α and IL-6 in serum （P<0.05）. The protein levels of LC3 and p-AMPK/AMPK in colon tissue were up-regulated， while the protein levels of p-mTOR/mTOR were down-regulated （P<0.01）. The mRNA expression levels of AMPK， Beclin1， and LC3 were up-regulated， while the mRNA expression of mTOR and p62 were down-regulated （P<0.01）. The colon tissue of the inhibitor group was severely damaged， and the trend of various indicators was completely opposite to that of the high dose paeoniflorin group. ConclusionPaeoniflorin can enhance autophagy and reduce inflammatory damage in mice with UC by activating the AMPK/mTOR signaling pathway and thus play a protective role.

Application of 18F-FDG PET in the diagnosis and evaluation of SARS-CoV-2 infection and related events / 中华核医学与分子影像杂志

Coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has rapidly ravaged the world and infected hundreds of millions of people since its outbreak. Early diagnosis of COVID-19 is conducive to the control of virus transmission and the timely treatment of patients. Among the imaging techniques, chest CT is an important basis for the diagnosis and evaluation of COVID-19. 18F-FDG PET is not generally recommended as a routine diagnostic tool for COVID-19, but it plays an important role in the assessment of SARS-CoV-2-related events based on the characteristic of whole-body multi-systemic scan and functional imaging diagnosis. In this paper, the application of 18F-FDG PET in COVID-19 diagnosis, prognostic evaluation and long-term sequelae evaluation, and clinical performance of 18F-FDG PET after COVID-19 vaccine are summarized on the basis of literature research and clinical reports analysis. Furthermore, the application and development direction of other new molecular probes for nuclear medicine in COVID-19 are prospected.

Preparation of Al 18F-PSMA-11 and its preliminary PET/CT imaging / 中华核医学与分子影像杂志

Objective:To optimize the preparation conditions and methods of Al 18F-prostate specific membrane antigen (PSMA)-11 and evaluate the feasibility of clinical transformation. Methods:PSMA- N, N′-bis(2-hydroxy-5-(carboxyethy)benzyl)ethylenediamine- N, N′-diacetic acid (HBED-CC) dissolved in CH 3COONH 4 buffer (pH=4.8) was reacted with AlCl 3·3H 2O dissolved in pure water at a molar ratio of 1∶1 (60 ℃, 10 min), and then purified by tC18 column and freeze-dried to obtain [Al]-PSMA-11. [Al]-PSMA-11 was labeled by 18F - and the effects of reaction temperature and pH value on the labeling rate were investigated. The labeled products were purified by tC18 column and filtered through sterile filter to obtain Al 18F-PSMA-11. The comparison of biodistribution between Al 18F-PSMA-11 and 68Ga-PSMA-11 was analyzed on 5 healthy volunteers (age (56±8) years). The differences of SUV max between two groups were analyzed by independent-sample t test. Besides, the early and delayed imaging of Al 18F-PSMA-11 PET/CT were performed on a patient (70 years old) with recurrent prostate cancer for assessment of its potential for prostate cancer recurrence monitoring. Results:The labeling rate was (42.3±3.2)% reacting in aqueous phase (60 ℃, pH=4.8) for 15 min. After being purified with tC18 cartridge, the radiochemical purity of the product was still more than 95% after placement at room temperature for 3 h. Preliminary application demonstrated that there was no significant difference in the biodistribution of Al 18F-PSMA-11 and 68Ga-PSMA-11 among lacrimal gland, parotid gland, submandibular gland, liver, spleen, kidney, bladder and part of intestine and SUV max of targeted organs were also not different ( t values: 0.19-1.95, all P>0.05) between two groups. Multiple bone metastases were observed by Al 18F-PSMA-11 PET/CT delayed imaging (3 h) in a patient with recurrent prostate cancer. Conclusion:Al 18F-PSMA-11 produced with pre-conjugated [Al]-PSMA-11 meets the requirement of the PET imaging application, and it has good potential of localization and imaging for prostate cancer metastatic lesions.

Serum LAPTM4B-35 protein as a novel diagnostic marker for hepatocellular carcinoma / 北京大学学报(医学版)

OBJECTIVE@#LAPTM4B-35 protein is one of the isoforms that are encoded by a cancer driver gene, LAPTM4B. This gene was primarily found and identified in our lab of Peking University School of Basic Medical Sciences. The LAPTM4B-35 protein and its encoded mRNA are significantly over-expressed in a variety of cancers, such as hepatocellular carcinoma (HCC), lung cancers (including non small-cell lung cancer and small-cell lung cancer), stomach cancer, colorectal carcinoma, pancreatic cancer, gallbladder cancer, cholangiocarcinoma, breast cancer, prostate cancer, ovarian cancer, cervical cancer, endometrial cancer, and so on. It has firmly demonstrated through lab experiments either in vivo or in vitro, as well as clinical studies that the over-expression of LAPTM4B-35 can promote cancer growth, metastasis, and multidrug resistance. Specially, the expressive level of LAPTM4B-35 is associa-ted with recurrence of HCC. The aim of this study is to identify the release of LAPTM4B-35 protein from hepatocellular carcinoma into blood of HCC patients and into the medium of cultured HCC cells, and to identify its possible form of LAPTM4B-35 protein existed in blood and cell culture medium, as well as to explore the possibility of LAPTM4B-35 protein as a novel HCC biomarker for diagnosis of HCC and prognosis of HCC patients.@*METHODS@#Immunobloting (Western blot) and enzyme-linked immunosorbent assay (ELISA) were used for identification of LAPTM4B-35 protein in the blood of HCC patients and normal individuals. Ultrafiltration and ultracentrifugation were used to isolate and purify exosomes from the culture medium of HCC cells.@*RESULTS@#LAPTM4B-35 protein existed in the blood from HCC patients and normal donors that were demonstrated through Western blot and ELISA. LAPTM4B-35 was also released into the culture medium of HCC cells in the form of exosomes. Preliminary experiments showed that the average and the median of LAPTM4B-35 protein level in the blood of HCC patients (n=43) were both significantly higher than that in the blood of normal donors (n=33) through sandwich ELISA.@*CONCLUSION@#It is promising that the LAPTM4B-35 protein which is released from HCC cells in the form of exosomes into their extraenvironment may be exploited as a novel cancer biomarker for HCC serological diagnosis.

FGF-2 combined with TGF-β1 induces bone marrow mesenchyma stem cells differentiating into cardio-myocyte-like cells in vitro / 实用医学杂志

Objective To expore the effects of FGF-2 and TGF-β1 on the differentiation of rat bone mar-row mesenchymal stem cells(BMSCs)towards cardiocytes. Methods BMSCs were isolated from bones of SD rats and cultured and identified by flow cytometry.The cells were divided into four groups:group A(FGF-2),group B (TGF-β1),group C(FGF-2+TGF-β1)and group D(blank control).The morphological changes were observed by the microscope. The expressions of Desmin,α-sarcomeric actin and cTnI were detected by immunocytochemical stainning.The expressions of GATA-4 and Nkx2.5was detected by RT-qPCR.Result The positive expression rate of CD90,CD29 and CD45 was 84.7%,86.5%,0.3% respectively. After induction,group C presence of myotube structures and the shape is fusiform.The positive rate of combined induction group was higher than the others (P < 0.05). The expression of GATA-4 and Nkx2.5 genes was higher than that in the other groups(P < 0.05). Conclution FGF-2 and TGF-β1 can be used to induce BMSCs into cardiocytes,and the combined group has the better effect.

Expression of ASPP2 and P16INK4a and its relationship with apoptosis in esophageal carcinoma / 实用医学杂志

Objective To study the expression of ASPP2 and P16INK4a in esophageal carcinoma and their relationship to the apoptosis and related clinicopathological characteristics. Methods Immunohistochemistry S-P method was used to examine the expression of ASPP2 and P16INK4a in the pathological specimens of 112 esophageal carcinoma cases and 31 cases of normal esophageal mucosa. TUNEL was also employed to detect the rate of apopto-sis in 37 esophageal carcinoma cases and 12 cases of normal esophageal mucosa. Results The difference between the expression of ASPP2 and P16INK4a in esophageal carcinoma and normal esophageal mucosa was statistically signif-icant(P < 0.05). Their abnormal expressions were all related to lymph node metastasis and differentiation degree (P < 0.05). The difference between the positive rate of apoptosis in esophageal carcinoma and normal esophageal mucosa was statistically significant(P < 0.05). The abnormal expression of ASPP2 and P16INK4a was all related to apoptosis in esophageal carcinoma. Conclusions The different expression of ASPP2 and P16INK4a may cooperatively play a role in differentiation degree ,lymph nodes metastasis and apoptosis in esophageal carcinoma. Co-examina-tion of them may be useful for the diagnosis and guiding the clinical treatment in esophageal carcinoma.

Expression of ASPP2 and P16INK4a and its relationship with apoptosis in esophageal carcinoma / 实用医学杂志

Objective To study the expression of ASPP2 and P16INK4a in esophageal carcinoma and their relationship to the apoptosis and related clinicopathological characteristics. Methods Immunohistochemistry S-P method was used to examine the expression of ASPP2 and P16INK4a in the pathological specimens of 112 esophageal carcinoma cases and 31 cases of normal esophageal mucosa. TUNEL was also employed to detect the rate of apopto-sis in 37 esophageal carcinoma cases and 12 cases of normal esophageal mucosa. Results The difference between the expression of ASPP2 and P16INK4a in esophageal carcinoma and normal esophageal mucosa was statistically signif-icant(P < 0.05). Their abnormal expressions were all related to lymph node metastasis and differentiation degree (P < 0.05). The difference between the positive rate of apoptosis in esophageal carcinoma and normal esophageal mucosa was statistically significant(P < 0.05). The abnormal expression of ASPP2 and P16INK4a was all related to apoptosis in esophageal carcinoma. Conclusions The different expression of ASPP2 and P16INK4a may cooperatively play a role in differentiation degree ,lymph nodes metastasis and apoptosis in esophageal carcinoma. Co-examina-tion of them may be useful for the diagnosis and guiding the clinical treatment in esophageal carcinoma.

Laminin and its alpha 6 integrin receptor in the regulation of human hepatocellular carcinoma cell phenotypes / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the role of integrin alpha 6 in cell attachment, spread, survival/proliferation and differentiation of human hepatocellular carcinoma (HCC) BEL-7402 cells on various substrates by monoclonal antibody against the extracellular domain of alpha 6 subunit (IA6ED McAb).</p><p><b>METHODS</b>The effect of McAb on attachment and spread of BEL-7402 cells on LN or FN substrate was examined. MTT analysis was used to examine the cell survival/proliferation, gelatin zymography to the matrix metalloproteinases (MMPs) secreted by BEL-7402 cells, and microparticle immunoabsorbent kit to AFP secretion while the cells were cultured on LN-, FN- or Matrigel-coated substrates.</p><p><b>RESULTS</b>The cell attachment, spread and survival/proliferation were inhibited. Moreover importantly, the malignant cell dedifferentiation and abnormal differentiation on LN-coated substrate were also strongly inhibited by IA6ED McAb.</p><p><b>CONCLUSION</b>LN and integrin alpha 6 regulate human HCC cell phenotypes of survival/proliferation and differentiation. The phenotypes of dedifferentiation and abnormal differentiation can be reversed by blocking the interaction between integrin alpha 6 and LN using IA6ED McAb, which may lower metastatic potency of tumor cells.</p>