AIM: Eight melanoma cell lines were establi sh ed from human melanoma specimen and designated as HME 1-HME 8, and their charact eristics of cell biology and immunology in vitro were studied. METHODS: The tissues from biopsies of human melanoma were cultur ed in RPMI 1640 media. After growing to 90% confluence, the cells were detached for subculture and then their characteristics, including morphology, growth kine tic, tumor antigen and tumorigencity in nude mice were studied. RESULTS: These cell lines have been passaged more than 100 times within 2 years. Their characteristics demonstrated: ① The population doubling time calculated in the log phase of growth were 34.2-59.5 h. ② The cloning ef ficiency in soft agar averaged 8.6%-26.2%. ③ The melanoma cells transplan t ed in nude mice were high tumorigencity. ④ Karyo-type analysis showed that aneu ploidy with the modal chromosomal number 64-117. ⑤ Rich ribosomes and melanoid grains in the cytoplasm were observed under electronic microscope. ⑥ Immunohist ochemistry analysis showed that there were a lot of brown grains in the cytoplas m. ⑦ Detection of tumor-antigen demonstrated that melan-A antigen was 75.0%, M AGE-1 was 50.0%, MAGE-2 was 37.5%, MAGE-3 was 62.5%, respectively, for eight melanoma cell lines. CONCLUSION: The melanoma cells have immortalized after being cul tured in vitro and has specific characteristics of malignant melanoma.

Objective To investigate the value and dynamic characteristics of diffusion-weighted imaging(DWI)in rabbit liver VX_2 tumor model,with correlation of pathology.Methods Forty New Zealand rabbits were included in the study and VX_2 tumor piece was implanted directly into the liver after laporotomy.Fiffteen had two intrahepatic implants while twenty-five had one implant.DWI was performed on the seventh,fourteenth and twenty-first day after implantation,while routine T_1WI and T_2WI sequences were done on the seventh and fourteenth day.Ten VX_2 tumor samples were studied by pathology.Results The lump detection rates on the seventh day after implantation of DWI,T_1WI and T_2WI were 78.7%(37/47), 10.7%(3/28)and 53.6%(15/28)respectively with statistical significance(x~2=32.61,P

PURPOSE: The purpose of this study was to detect the lymphatic drainage pattern of internal mammary area and verify the concept of internal mammary sentinel lymph node (IM-SLN) in breast. MATERIALS AND METHODS: A small particle radiotracer ((99m)Tc-Dextran 40) was prepared and tested. (99m)Tc-Dextran 40 was injected into intraparenchyma at the sound breast by a modified radiotracer injection technique. Subsequently, dynamic single-photon emission computed tomography (SPECT), computed tomography (CT), and SPECT/CT combination images were performed to identify the radioactive lymph vessels and internal mammary lymph nodes (IMLNs). The direction of lymph drainage and the location of the IMLNs were identified in the SPECT/CT imaging. RESULTS: The radiochemical purity of (99m)Tc-Dextran 40 was > 95%. (99m)Tc-Dextran 40 could drainage into first, second, and third lymph node and the radioactive lymph node could be detected by the γ detector in the animal experiment. After (99m)Tc-Dextran 40 injecting into intraparenchyma, 50.0% cases (15/30) were identified the drainage lymphatic vessels and radioactive IMLNs by SPECT. The drainage lymphatic vessel was found from injection point to the first IMLN (IM-SLN) after 10.5±0.35 minutes radiotracer injection, and then (99m)Tc-Dextran 40 was accumulated into the IM-SLN. The combination imaging of SPECT/CT showed the second IMLN received the lymph drainage from the IM-SLN. The lymphatic drainage was step by step in the internal mammary area. CONCLUSION: The lymph was identified to drain from different regions of the breast to IM-SLN, and then outward from IM-SLN to other IMLN consecutively. It demonstrated the concept of the IM-SLN and provided more evidences for the application of internal mammary sentinel lymph node biopsy.
Animal Experimentation ; Breast Neoplasms ; Breast ; Drainage ; Lymph Nodes ; Lymphatic Vessels ; Sentinel Lymph Node Biopsy ; Tomography, Emission-Computed ; Tomography, Emission-Computed, Single-Photon

OBJECTIVETo search for an appropriate animal model that is more closely related to human to study cAMP-response element binding protein target gene Staufen and to identify its location.

METHODSThe phylogenetic tree was constructed with Staufen protein (STAUFEN) sequences of different species, and the most suitable animal model was selected by analyzing relativity among them. The Staufen fragments were amplified with reverse transcription-PCR and inserted into a vector and then the sub-clone was transformed into bacteria, selected, amplified, extracted and sequenced. Staufen probes were in vitro transcribed and hybridized in situ on the cryosections of the mouse brain. The cryosections were stained and observed.

RESULTSThe clustering patterns of the phylogenetic tree indicated that the mouse and human Staufen1 had 99.7% protein sequences similarity. The mRNA of Staufen was located in CA1, CA2, CA3 and DG hippocampus regions shown by in situ hybridization.

CONCLUSIONThe mouse is a preferable animal model for research on Staufen transcription in hippocampus.

Animals ; Cyclic AMP Response Element-Binding Protein ; metabolism ; Hippocampus ; metabolism ; In Situ Hybridization ; Mice ; RNA, Messenger ; genetics ; RNA-Binding Proteins ; genetics ; metabolism

BACKGROUNDTo observe cytopathogenic effect of Hantaan virus (HV) on cultured human bone marrow cells.

METHODSLight and transmission electron microscopy and direct immunofluorescent technique were applied to study cellular structure especially ultrastructural changes of bone marrow cells from patients with Hantaan virus infection. Bone marrow cells of one healthy volunteer were also studied as control.

RESULTSThe antigen of HV was found in bone marrow cells of 20 of 27 HFRS patients by the aid of direct immunofluorescent technique. It was found that the granulocytes had the highest percentage of HV antigen positive cells (76%), followed by monocytes (65%), lymphocytes (40%), megakaryocytes (20%) and the lowest was found in erythrocytes (3.7%). The injury of cell membrane after infection with HV was significantly more severe than that in the control group under the light and electron microscopy.

CONCLUSIONThis study demonstrated that HV could attack human bone marrow cells and cause cytopathogenic effect on them.

Adult ; Aged ; Antigens, Viral ; analysis ; Bone Marrow Cells ; ultrastructure ; virology ; Female ; Fluorescent Antibody Technique, Direct ; Hantavirus ; immunology ; pathogenicity ; Hemorrhagic Fever with Renal Syndrome ; pathology ; virology ; Humans ; Male ; Microscopy, Electron, Scanning ; Microscopy, Fluorescence ; Middle Aged

Objective To investigate the influencing factors on behavior related to HIV testing among female commercial sex workers under Structural Equation Model (SEM).Methods In Chongqing,Sichuan,Guangxi,Xinjiang provinces,1613 female commercial sex workers were participated in a questionnaire survey.Factors on behaviors related to HIV testing among female commercial sex workers were analyzed based on SEM.Results Influencing factors on behaviors related to HIV testing among female commercial sex workers would include social status,knowledge on AIDS,risk through self-evaluation,condom use,frequency of sexual services etc.GFI,AGFI,RMR were 0.9952,0.9898 and 0.0115 respectively.Conclusion Social status,knowledge on AIDS,risk through self-evaluation,condom use and frequency of sexual services were affecting the behaviors related to HIV testing among female commercial sex workers.

Objective:To study combined adjuvant transcatheter arterial chemoembolization (TACE) with anti-tumor drug treatment on early hepatocellular carcinoma (HCC) recurrence in patients with microvascular invasion (MVI) after partial hepatectomy with curative intent.Methods:The clinical and pathological data of 169 patients with HCC who underwent partial hepatectomy with curative intent from January 2015 to December 2018 at the Affiliated Hospital of Qingdao University were retrospectively analyzed. MVI was diagnosed by postoperative histopathology. There were 147 males and 22 females, with the median age 56 years(ranged 32-79 years). The patients were divided into surgery group ( n=62, patients who did not receive adjuvant therapy), TACE group ( n=42, patients who only received TACE) and combined group ( n=65, patients who received TACE with anti-tumor drug) according to the therapies after resection. Patients in each group were further divided into grade M1 (mild) and grade M2 (severe) subgroups according to the severity of MVI. All patients were followed-up for observing tumor recurrence. The relapse-free survival in the three groups were compared using the Kaplan-Meier method and the log-rank test was used to compare the tumor-free survival rates. Results:The tumor-free survival rates of 169 patients at 1 and 2 years after operation were 59.2% and 40.8%. The tumor-free survival rates at 1 and 2 years after operation were 45.2% and 25.8% in surgery group, 61.9% and 40.5% in TACE group, 70.8% and 52.3% in combined group respectively. The differences among the three groups were significant: TACE group was better than surgery group, and combined group was better than TACE group, combined group was better than surgery group (all P<0.05). In TACE group and combined group, tumor-free survival rates of M1patients better than M2 patients, and the difference was significant ( P<0.05). Among M1 patients and M2 patients, tumor-free survival rates of combined group patients were better than surgery group and TACE group, the difference was significant (all P<0.05). The cumulative tumor-free survival rate was not significantly affected by different antineoplastic agents. Conclusion:Adjuvant TACE reduced the early recurrence rate of HCC patients with MVI. Adjuvant TACE combined with anti-tumor drug further reduced early tumor recurrence.

Objective:To analyze the distribution of pathogenic bacteria in bile culture in patients with common bile duct stones and biliary tract infections, in order to guide clinical optimization of antibiotics application.Methods:From March 30, 2017 to December 31, 2021, at Affiliated Hospital of Qingdao University, 753 patients with common bile duct stones and biliary tract infections and received endoscopic retrograde cholangiopancreatography were selected. Bile samples were obtained for bacterial culture, strain type identification and drug sensitivity test in order to analyze bile pathogenic bacteria distribution, change trend and drug resistance. Chi-square test was used for statistical analysis.Results:From 2017 to 2021, the total positive rate of bile culture in 753 patients with choledocholithiasis complicated with biliary tract infection was 90.17% (679/753). From 2017 to 2021, the positive rates of bile culture were 82.05% (64/78), 88.81% (119/134), 88.03% (125/142), 93.87% (199/212), and 91.98% (172/187), respectively, and the difference was statistically significant ( χ2=10.78, P=0.029). The positive rate of bile culture in 2017 was lower than those in 2020 and 2021, and the differences were statistically significant ( χ2=9.43 and 5.57, P=0.002 and 0.018). There were no significant differences in the positive rates of bile culture among the other years (all P>0.05). A total of 1 033 pathogenic bacteria were detected in the 679 bile specimens with positive bile culture results. Among which the total proportion of Gram-negative bacilli was 57.02% (589/1 033), and from 2017 to 2021 the proportions were 66.38% (77/116), 66.47% (111/167), 59.43% (104/175), 54.75% (173/316), and 47.88% (124/259), respectively. The total proportion of Gram-positive cocci was 41.05% (424/1 033), and from 2017 to 2021 the proportions were 31.90% (37/116), 31.74% (53/167), 38.86% (68/175), 44.30% (140/316), and 48.65% (126/259), respectively. The total proportion of fungus was 1.94% (20/1 033), and from 2017 to 2021 the proportions were 1.72% (2/116), 1.80% (3/167), 1.71% (3/175), 0.95% (3/316), and 3.47% (9/259), respectively. From 2017 to 2021, the proportion of Gram-negative bacilli gradually decreased, while the proportion of Gram-positive cocci gradually increased, and the differences were statistically significant ( χ2=20.14 and 17.91, P<0.001 and =0.001). From 2017 to 2021, the change in the proportion of fungus was not statistically significant ( P>0.05). The main Gram-negative bacilli in the bile culture were Escherichia coli (31.36%, 324/1 033) and Klebsiella pneumoniae (12.68%, 131/1 033); the main Gram-positive cocci were Enterococcus faecalis (14.04%, 145/1 033) and Streptococcus salivarius (4.36%, 45/1 033). From 2017 to 2021, the proportions of Escherichia coli were 39.66% (46/116), 38.92% (65/167), 33.14% (58/175), 28.48% (90/316), and 25.10% (65/259), respectively, with gradual decrease and the difference was statistically significant ( χ2=14.34, P=0.006). From 2017 to 2021 the detection rates of extended-spectrum β-lactamase (ESBL) in Escherichia coli and Klebsiella pneumoniae were 30.43% (14/46), 26.15% (17/65), 29.31% (17/58), 38.89% (35/90), 40.00% (26/65), and 4/15, 20.00% (5/25), 20% (5/25), 24.32% (9/37), and 31.03% (9/29), and there were no significant differences in the detection rates of ESBL between different years (both P>0.05). Conclusions:From 2017 to 2021, the positive rate of bile culture in patients with choledocholithiasis complicated with biliary tract infection showed an overall increasing trend. Gram-negative bacilli were still dominated in bile pathogenic bacteria, while the proportion of Gram-positive cocci remarkably increased, and the bile bacterial spectrum significantly changed. Clinicians should adjust the antibiotic dosing regimens according to the variation of bacterial spectrum and drug resistance.

OBJECTIVETo compare the efficacy and safety of subcutaneous immunotherapy with dermatophagoides pteronyssinus standardized extract given in conventional and cluster immunotherapy schedules for persistent allergic rhinitis.

METHODSOne hundred and ten patients with moderate to severe allergic rhinitis caused by dust mites, in accordance with the immunotherapy inclusion criteria, were allocated to receive conventional immunotherapy as group A (n = 57) or cluster immunotherapy as group B (n = 53). In group A, 7 cases were lost to follow-up, the expulsion rate of group A was 12.28%; in group B, 1 case was lost to follow-up, the expulsion rate of group B was 1.89%. Nasal symptom scores, medicine scores and mini rhinoconjunctivitis quality of life questionnaire (Mini RQLQ) were recorded and compared before and after 7 weeks, 15 weeks, 1.0 year, 1.5 years, 2.0 years. All the scores were assessed to evaluate the clinical efficacy, and also the incidence of local and systemic adverse reactions were registered to evaluate the safety. SPSS 19.0 software was used to analyze the data.

RESULTSNasal symptom scores, medicine scores and Mini RQLQ of both groups were significant lower than those before the treatment (all P < 0.05). Mini RQLQ and nasal symptom scores in cluster group (0.55 ± 0.21,0.57 ± 0.27) were more significantly declined than the conventional group after 7 weeks and 2.0 years of observation (all PMini RQLQ<0.05;nasal symptom scores: 1.41 ± 0.65, 0.83 ± 0.30, t value was 11.344, 5.649, both P < 0.05). The clinical efficiency rate in cluster group (86.5%, 94.2%) were more significantly highter than those (60.0%, 80.0%) in the conventional group after 7 weeks and 2 years of observation (χ(2) value was 9.224, 4.642, both P < 0.05). The medicine scores in cluster group (0.11 ± 0.04) was more significantly declined than conventional group (0.47 ± 0.11) after 7 weeks (t = 27.665, P < 0.05). The incidence of local and systemic adverse reactions during the incremental-dose phase and maintenance-dose phase compared with conventional immunotherapy were not significantly different (P > 0.05).

CONCLUSIONThe cluster immunotherapy is a safe treatment method which is more effective and faster than conventional immunotherapy to the dust mites caused allergic rhinitis.

Animals ; Dermatophagoides pteronyssinus ; Female ; Humans ; Immunotherapy ; Lost to Follow-Up ; Male ; Medicine ; Pyroglyphidae ; Quality of Life ; Reference Standards ; Rhinitis, Allergic ; immunology ; therapy ; Safety ; Surveys and Questionnaires

Berberine(BBR)is an isoquinoline alkaloid extracted from Coptis chinensis that improves diabetes,hyperlipidemia and inflammation.Due to the low oral bioavailability of BBR,its mechanism of action is closely related to the gut microbiota.This study focused on the CYP51 enzyme of intestinal bacteria to elucidate a new mechanism of BBR transformation by demethylation in the gut microbiota through multiple analytical techniques.First,the docking of BBR and CYP51 was performed;then,the pharma-cokinetics of BBR was determined in ICR mice in vivo,and the metabolism of BBR in the liver,kidney,gut microbiota and single bacterial strains was examined in vitro.Moreover,16S rRNA analysis of ICR mouse feces indicated the relationship between BBR and the gut microbiota.Finally,recombinant E.coli con-taining cyp51 gene was constructed and the CYP51 enzyme lysate was induced to express.The metabolic characteristics of BBR were analyzed in the CYP51 enzyme lysate system.The results showed that CYP51 in the gut microbiota could bind stably with BBR,and the addition of voriconazole(a specific inhibitor of CYP51)slowed down the metabolism of BBR,which prevented the production of the demethylated metabolites thalifendine and berberrubine.This study demonstrated that CYP51 promoted the deme-thylation of BBR and enhanced its intestinal absorption,providing a new method for studying the metabolic transformation mechanism of isoquinoline alkaloids in vivo.