Objective To investigate the impact of serine/threonine-protein kinase 1(AKT1)-mediated autophagy in hepatocellular carcinoma(HCC)cells on their sensitivity to 125I seed irradiation.Methods ① iProX database and STRING12.0 website were utilized to analyze the proteomic data of HCC before and after 125I seed irradiation to explore the differentially expressed proteins and associated functional connections.Meanwhile,The Cancer Genomics Atlas(TCGA)database was employed to analyze the relationship between AKT1 expression level and the survival of HCC patients.② Human HCC cell lines HUH7 and Hep3B were exposed to continuous irradiation from 125I radioactive seeds with an initial apparent activity of 0.8 mCi per seed for approximately 120 h,accumulating a total dose of 8 Gy,while the control cells were cultured under normal condition for 120 h.③ Autophagy inhibitor,chloroquine(CQ)and inducer,rapamycin(RAPA)were used to treat the HCC cells respectively to establish the CQ group and the RAPA group.The lentiviral transfection technique was employed to construct the HCC cells with overexpressed AKT1,namely the AKT1 group.The HCC cells treated in the same way were continuously irradiated with 125I seeds for 120 h to construct the CQ+125I group,the RAPA+125I group,and the AKT1+125I group.④The changes in microtubule-associated protein light chain 3(LC3),p62,AKT1 and p-AKT1 were detected by Western blotting.Cell immunofluorescence assay was employed to observe the expression of autophagy related proteins,such as LC3.The colony forming ability and apoptotic rate were detected with plate cloning assay and flow cytometry.Results ① Continuous irradiation with 125I seeds resulted in decreased expression of p62 and increased ratio of LC3Ⅱ/LC3Ⅰ(P＜0.05)when compared with the negative control(NC)group.Immunofluorescence assay revealed more green fluorescence spots of LC3.When compared with the 125I group,the CQ+125I group had significantly increased expression of p62(P＜0.01),decreased ratio of LC3Ⅱ/LC3Ⅰ(P＜0.01),lower apoptotic rate(P＜0.01),and more colony formations(P＜0.01).In contrast,the results in the RAPA+125I group were opposite to those of the CQ+125I group.② Analysis on the iProX database showed that the expression of AKT1 was decreased in the irradiated group,and analysis on the TCGA database indicated that high expression of AKT1 predicted a poor prognosis for HCC patients(P＜0.01).③After irradiation with 125I seeds,the expression of AKT1 at both the RNA and protein levels was decreased in the 125I group(P＜0.01).After overexpression of AKT1,the level of autophagy was decreased(P＜0.05).Irradiation of HCC cells with overexpressed AKT1 using 125I seeds could partially restore the level of autophagy.In the AKT1+125I group,the expression of AKT1,pAKT1 and p62 were all decreased,and the ratio of LC3Ⅱ/LC3Ⅰwas increased than the AKT1 group(P＜0.05).④ The apoptotic rate of the AKT1+125I group was lower than that of the 125I group(P＜0.05)and higher than that in the AKT1 group(P＜0.05).In HUH7 cells,the clonogenic ability of the AKT1+125I group was higher than that of the 125I group(P＜0.05).In Hep3B cells,the clonogenic ability of the AKT1+125I group was higher than that of the 125I group,and the clonogenic ability of the AKT1 group was higher than that of the NC group(P＜0.05).Conclusion 125I seed irradiation induce lethal autophagy in HCC cells by reducing the expression of AKT1,providing a new theoretical basis for the implantation of 125I radioactive seeds in the treatment of HCC.

Objective To investigate the advantages and disadvantages of percutaneous bone marrow puncture biopsy guided by automatic laser navigation and free-hand percutaneous bone marrow biopsy guided by CT.Methods Seventy patients with bone marrow biopsy at the department of nuclear medicine in this hos-pital from January to June 2022 were selected as the study subjects and divided into the laser navigation group (bone marrow biopsy guided by automatic laser navigation,n=30) and free-hand puncture group (percutane-ous bone marrow biopsy guided by CT,n=40) based on different guidance methods.The duration of the pro-cedure,total radiation dose,total number of scans,positive rate of bone lesion biopsy and puncture complica-tions occurrence were compared between the two groups.Results Compared to the free-hand puncture group,the operation duration in the laser navigation group was shorter[11.00(7.00,13.00)min vs. 13.00(11.00,17.75)min],the total radiation dose was lower[439.00(291.00,568.75)mGy vs. 510.00(431.25,708.50) mGy],total number of scans was less[4.50(3.00,6.00)times vs. 6.00(5.00,7.00)times],and the differ-ences were statistically significant (P<0.05).There was no statistically significant difference in the positive rate of bone lesion biopsy between the laser navigation group and free-hand puncture group[96.67％(29/30) vs. 85.00％(34/40),P>0.05].No puncture complications occurred in both groups.Conclusion Automatic laser navigation could assist the operators to better complete percutaneous bone marrow biopsy procedures.

Objective To investigate the effect of progesterone receptor membrane component 1(PGRMC1)mediated autophagy on the sensitivity of liver cancer cells to 125I particles irradiation.Methods Hepatoma cell lines Huh7 and LM3 were exposed to different doses(0,2,4,6 and 8 Gy)of 125I particles,and cell autophagy was observed by transmission electron microscopy(TEM).Then,autophagy inhibitor chloroquine(CQ),agonist rapamycin(Rapa),and PGRMC1 inhibitor AG-205 were used respectively to verify that PGRMC1-mediated autophagy plays a key role in the sensitivity of hepatocellular carcinoma cells to 125I particle irradiation.Cell proliferation,colony formation and apoptosis were detected by CCK-8 assay,clonal formation test and flow cytometry,respectively.The expression levels of PGRMC1,microtubule-associated protein light chain 3-Ⅰ(LC3-Ⅰ),LC3-Ⅱ and p62 were detected by Western blotting.Results Different doses of 125I particles irradiation significantly decreased the proliferation and clonogenesis of Huh7 and LM3 cells(P＜0.05),and increased the apoptotic cells(P＜0.01),in a dose-dependent manner.Compared with the 0 Gy group,the ratio of LC3-Ⅱ/LC3-Ⅰ in Huh7 and LM3 cells was obviously increased,and the expression of p62 was significantly down-regulated in the 6 Gy group.The proliferation capacity and clonal formation ability of Huh7 and LM3 cells were decreased significantly,and their apoptotic cells were increased notably in the 6 Gy+CQ group than the 6 Gy group,while the above results were on the contrary in the 6 Gy+Rapa group.The 6 Gy+AG205 group had notably decreased LC3-Ⅱ/LC3-Ⅰ ratio in the Huh7 and LM3 cells,up-regulated p62 expression,reduced cell proliferation capacity and clone formation ability,and enhanced cell apoptosis when compared with the 6 Gy group,and the above results of the 6 Gy+PGRMC1 group were opposite.Conclusion Increment of PGRMC1 induced by 125I irradiation can promote autophagy,increase the proliferation and clonogenesis,and reduce the apoptosis in hepatocellular carcinoma cells.

Objective:To evaluate the efficacy and prognostic factors of 125I seeds implantation for primary hepatocellular carcinoma. Methods:From December 2011 to January 2021, 102 primary hepatocellular carcinoma patients (86 males, 16 females; median age 61 years) who underwent 125I seeds implantation from 5 hospitals in China were enrolled in this retrospective study. Local progression-free survival (LPFS), overall survival (OS) and the prognostic factors were analyzed. Kaplan-Meier method was used to draw the distribution curve of survival time, and LPFS rate and OS rate were calculated. Log-rank test and Cox regression were used to analyze the influencing factors of survival. Results:The median follow-up time was 38 months until April 2021. The local control rate was 96.1%(98/102). The 1-, 3- and 5-year LPFS rate were 61.3%, 25.5% and 12.7%, and the 1-, 3- and 5-year OS rate were 73.9%, 39.1% and 22.6%, respectively. There were 75 patients with progressive disease, including 42 patients with intrahepatic recurrence and metastasis after seed implantation, and 55 patients died. Multivariate analyses showed that short-term efficacy complete response (CR) (hazard ratio ( HR)=0.34, 95% CI: 0.20-0.58) was protective factor related to LPFS; short-term efficacy CR ( HR=0.25, 95% CI: 0.13-0.47) was the protective factors related to OS; Barcelona clinic liver cancer (BCLC) C stage ( HR=2.33, 95% CI: 1.27-4.27), intrahepatic progression and extrahepatic metastasis ( HR=3.18, 95% CI: 1.28-7.86; HR=3.23, 95% CI: 1.27-8.21) were independent risk factors related to OS. No sever adverse effects were observed. Conclusions:125I seeds implantation is safe and effective for the treatment of primary hepatocellular carcinoma. BCLC stage, short-term efficacy and post-implantation progression are independent factors related to survival time.

Objective:To compare the differences in dosimetric parameters of target areas between 125I seed implantation with degradable catheters and by hand under the assistance of a real-time intraoperative treatment planning system (TPS). Methods:Forty-two simulated lesions were divided into a degradable catheter group and a free-hand group, with twenty-one lesions in each group. 125I seeds were implanted according to the TPS.The pre-plan and post-implant dosimetric parameters were collected, including the minimum dose ( Dmin), maximum dose ( Dmax), mean dose ( Dmean), conformal index (CI), external index (EI), homogeneity index (HI), minimum prescription dose delivered to 90% of the target volume ( D90), and the mean percentage of volume receiving 90% of the prescription doses ( V90). The Bland-Altman method was employed to analyze the consistency of pre-plan and post-implant dosimetric parameters and the Wilcoxon rank-sum test was used for the comparison of the two groups. Results:According to the Bland-Altman analysis, the dosimetric parameters of the two groups were all in agreement before and after seed implantation except for the Dmin and V90 of the free-hand group.Furthermore, the degradable catheter group had smaller error ranges of Dmax ( Z=-3.824, P<0.005), CI ( Z=-1.962, P<0.005), HI ( Z=-2.352, P<0.005), D90 ( Z=-2.453, P<0.005), and V90 ( Z=-3.159, P<0.005). Conclusions:The dosimetric parameters of 125I seed implantation with degradable catheters under the assistance of real-time TPS have good pre-plan and post-implant consistency and smaller error ranges.

Objective:Through the establishment and application of the biosafety autopsy pathology platform in Huoshenshan Hospital, the feasibility and application effect of the biosafety autopsy pathology platform were analyzed.Methods:The feasibility and application effect of the biosafety autopsy pathology platform were analyzed by layout design, instruments and equipment preparation, testing methods examination, and effect evaluation.Results:A total number of 26 cases of systematic autopsy and 8 cases of minimally invasive autopsy (puncture) were performed on the biosafety autopsy pathology platform, and no one was infected. Some pathology original findings were identified, including COVID-19 and pathological characteristics of identification, SARS respiratory failure mechanism and treatment significance, systemic distribution and spreading mechanism of the new coronavirus, the " storm" of inflammation pathological basis, some tumor markers rise in pulmonary pathological cell source and the overcast with fibrosis characteristics, such findings play important roles in the clinical diagnosis and treatment of COVID-19.Conclusions:The study of autopsy pathology is of great significance for the prevention and control of emerging infectious disease, which calls for early intervention. To promote the standard construction of biosafety autopsy platform is the key to the pathological study of emerging infectious diseases. Pathological research and clinical diagnosis and treatment should be combined to inform each other.

Objective:To investigate the effect of radioactive 125I seed on angiogenesis of subcutaneously transplanted hepatocellular carcinoma in nude mice and underlying mechanism. Methods:The subcutaneous transplanted tumor model of human hepatocellular carcinoma Huh7 cells was established in nude mice. Twelve nude mice were randomly divided into observation group and control group with 6 mice in each group. The 125I seed with activity of 2.96×10 7Bq was implanted into the transplanted tumor of observation group and another with 0 Bq as control group, respectively. The volume of the transplanted tumor was measured every 4 d and the growth curve of the tumor was recorded. The microvessel density (MVD) of the transplanted tumor was evaluated by immunohistochemical detection of CD31. VEGF-A and HIF-1α protein and mRNA were detected by immunohistochemistry and RT-PCR, respectively. Results:The growth rate of tumor in the observation group was slower than that in the control group, and the difference of tumor volume between two groups at 12 d after 125I seed implantation was significantly different( t=3.167, P<0.05). At 28 d after transplantation, the tumor volumes of control and observation group approached to (963.61 ± 89.56) mm 3and (602.10±75.98) mm 3, respectively. The MVD of the observation group was significantly lower than that of the control group ( t=6.361, P<0.05). The relative expression of VEGF-A and HIF-1α mRNA in the observation group was significantly lower than that in the control group ( t=10.480, 6.414, P<0.05). Protein expression levels of VEGF-A and HIF-1α in the observation group were lower than those in the control group ( t=10.890, 12.250, P<0.05). Conclusions:Radioactive 125I seed can inhibit the growth of HCC xenografts by reducing tumor microvessels, which may be related to the decrease of VEGF-A and HIF-1α expression.

Objective:To investigate the pathological characteristics and the clinical significance of novel coronavirus (2019-nCoV)-infected pneumonia (termed by WHO as coronavirus disease 2019, COVID-19).Methods:Minimally invasive autopsies from lung, heart, kidney, spleen, bone marrow, liver, pancreas, stomach, intestine, thyroid and skin were performed on three patients died of novel coronavirus pneumonia in Chongqing, China. Hematoxylin and eosin staining (HE), transmission electron microcopy, and histochemical staining were performed to investigate the pathological changes of indicated organs or tissues. Immunohistochemical staining was conducted to evaluate the infiltration of immune cells as well as the expression of 2019-nCoV proteins. Real time PCR was carried out to detect the RNA of 2019-nCoV.Results:Various damages were observed in the alveolar structure, with minor serous exudation and fibrin exudation. Hyaline membrane formation was observed in some alveoli. The infiltrated immune cells in alveoli were majorly macrophages and monocytes. Moderate multinucleated giant cells, minimal lymphocytes, eosinophils and neutrophils were also observed. Most of infiltrated lymphocytes were CD4-positive T cells. Significant proliferation of type Ⅱ alveolar epithelia and focal desquamation of alveolar epithelia were also indicated. The blood vessels of alveolar septum were congested, edematous and widened, with modest infiltration of monocytes and lymphocytes. Hyaline thrombi were found in a minority of microvessels. Focal hemorrhage in lung tissue, organization of exudates in some alveolar cavities, and pulmonary interstitial fibrosis were observed. Part of the bronchial epithelia were exfoliated. Coronavirus particles in bronchial mucosal epithelia and type Ⅱ alveolar epithelia were observed under electron microscope. Immunohistochemical staining showed that part of the alveolar epithelia and macrophages were positive for 2019-nCoV antigen. Real time PCR analyses identified positive signals for 2019-nCoV nucleic acid. Decreased numbers of lymphocyte, cell degeneration and necrosis were observed in spleen. Furthermore, degeneration and necrosis of parenchymal cells, formation of hyaline thrombus in small vessels, and pathological changes of chronic diseases were observed in other organs and tissues, while no evidence of coronavirus infection was observed in these organs.Conclusions:The lungs from novel coronavirus pneumonia patients manifest significant pathological lesions, including the alveolar exudative inflammation and interstitial inflammation, alveolar epithelium proliferation and hyaline membrane formation. While the 2019-nCoV is mainly distributed in lung, the infection also involves in the damages of heart, vessels, liver, kidney and other organs. Further studies are warranted to investigate the mechanism underlying pathological changes of this disease.

Objective: To investigate the safety and efficacy of irreversible electroporation (IRE) hepatic ablation with high-frequency bipolar pulse in swine. Methods: The experimental study was conducted. A total of 18 swines of either gender, aged (6.8+ 0.8)months with a range of 5.5-8.0 months, were collected from Animal Laboratory Center of Army Medical University. were randomly divided into 15 in experimental group and 3 in control group. The swines in experimental group underwent IRE hepatic ablation with high-frequency bipolar pulse, and 3 swines were chose randomly and underwent enhanced CT examination immediately after ablation, and at 3, 7, 14, and 28 days after ablation. The liver tissues were taken for histopathological examination. The swines in the control group underwent IRE hepatic ablation with high-frequency monopolar burst, and was performed enhanced CT examination at 3 days after ablation. Liver tissues were taken for histopathological examination. Observation indicators: (1) comparison of muscle contraction of siwnes between two groups; (2) imaging performance on enhanced CT after IRE ablation in the experimental group; (3) hepatic histopathological findings after IRE ablation in the experimental group; (4) comparison of apoptotic index in the ablation zone between two groups. The measurement data with normal distribution were expressed as Mean±SD, and comparison between groups was performed by the independent sample t test. Results: (1) Comparison of muscle contraction between two groups: swines in both groups underwent ablation successfully. The degree of muscle contraction was (9.8±0.4)m/s² and (48.6±0.5)m/s² in the experimental group and in the control group, respectively, showing statistically significant difference between the two groups (t=-163.50, P<0.05). (2) Imaging performance on enhanced CT after IRE ablation in the experimental group: the enhanced CT examination of swines immediately after IRE ablation showed a low-density shadow and clear boundary in the ablation zone. There was no obvious abnormality in the ablation zone and its adjacent large vessels. No serious complications occurred after the ablation. The boundary between the ablation zone and the normal liver tissue of the experimental group gradually became blurred over time, and the ablation zone was gradually replaced by normal liver tissue. The ablation zone at the 28 days after ablation was significantly reduced or even disappeared on imaging of enhanced CT examination.The maximum diameter of the ablation zone was (1.81±0.17)cm immediately after ablation, (1.75±0.19)cm at the 3 days after ablation, (1.32±0.22)cm at the 7 days after ablation, (0.65±0.14)cm at the 14 days after ablation, (0.28±0.10)cm at the 28 days after ablation, respectively. (3) Hepatic histopathological findings after IRE ablation in the experimental group: the HE staining of ablated tissue immediately after ablation showed that the cells in the ablation zone were swollen, arranged disorderly, and bleeding was observed around some of the needles.The bile ducts and blood vessels were intact in the ablation zone, and a large number of deeply stained nuclei were seen at 3 days after ablation, some of the nucleus and apoptotic bodies were partially dissolved or cleaved. A large number of inflammatory cell were infiltrated around the ablation zone. Intact vascular and biliary endothelial cells were observed by von Willebrand factor staining, a larger number of apoptotic cells with deeply stained nuclei in the ablation zone were observed by terminal-deoxynucleoitidyl transferase mediated nick end labeling staining, and partial deposited dark brown calcium salt was seen by Von Kossa staining. More newborn hepatocytes were observed growing from the periphery of the ablation zone to the center at the 7, 14, 28 days after ablation. Smooth muscle cell proliferation was observed at 14 and 28 days after ablation. The ablation zone was replaced by new cells on 28 days after ablation. (4) Comparison of apoptotic index in the ablation zone between two groups: the apoptotic index of the ablation zone was significantly higher in the experimental group than in the control group on the 3 days after operation (76.67%±0.04% vs. 64.03%±0.05%, t=4.79, P<0.05). Conclusion IRE hepatic ablation of swine using high-frequency bipolar pulse is safe and reliable, and it has more apoptotic cells than IRE ablation with high-frequency monopolar burst.

Objective To compare the accuracy of 125I seeds longitudinal distance between groups by degradable catheters and hand.Methods The study was divided into two groups (implantation by degradable catheters and that by hand).There were 12 simulated lesions in each group.Seed needle and 125I seeds were implanted by three physicians according to the set longitudinal distance (0.5 and 1.0 cm).Postoperative CT was conducted and the longitudinal distance between seeds was measured,and then the data was analyzed between the actual distance and the designed distance in each group.Results The degree of deviation from the set distance were (0.184 ±0.047) and (0.127 ±0.051) cm in the group by hand,and (0.007 ± 0.006) and (-0.003 ± 0.006) cm in degradable catheters group.The degree of deviation of the seeds in the catheter group was significantly lower than the group by hand (t =3.804,2.499,P ＜0.05).Conclusions The accuracy of the longitudinal distance of the group by catheter is much better than that by hand.