Objective: To explore a simple and feasible method for the isolation and purification of hepatocytes, hepatic stellate cells (HSC), and lymphocytes from mice. Methods: The cell suspension was obtained from male C57bl/6 mice by hepatic perfusion through the portal vein digestion method and then isolated and purified by discontinuous Percoll gradient centrifugation. Trypan blue exclusion was used to determine cell viability. Glycogen staining, cytokeratin 18, and transmission electron microscopy were used to identify hepatic cells. Immunofluorescence was used to detect α-smooth muscle actin combined with desmin in HSCs. Flow cytometry was used to analyze lymphocyte subsets in the liver. Results: After isolation and purification, about 2.7×10(7) hepatocytes, 5.7×10(5) HSCS, and 4.6×106 hepatic mononuclear cells were obtained from the liver of mice with a body weight of about 22g. The cell survival rate in each group was > 95%. Hepatocytes were apparent in glycogen deposited purple-red granules and cytokeratin 18. Electron microscopy showed that there were abundant organelles in hepatocytes and tight junctions between cells. HSC had expressed α-smooth muscle actin and desmin. Flow cytometry showed hepatic mononuclear cells, including lymphocyte subsets such as CD4, CD8, NKs, and NKTs. Conclusion: The hepatic perfusion through the portal vein digestion method can isolate multiple primary cells from the liver of mice at once and has the features of simplicity and efficiency.
Male ; Mice ; Animals ; Keratin-18 ; Actins ; Desmin ; Liver ; Hepatocytes ; Hepatic Stellate Cells

Objective: To understand the characteristics and depressive status of men who have sex with men (MSM) who self-reported HIV infection through heterosexual transmission and to provide evidence for personalized management of HIV infected people. Methods: A cross-sectional survey was carried out in Shandong province from September to December in 2019. Male HIV patients aged ≥15 years who self-reported HIV infection through heterosexual transmission were selected to verify the route of infection by one-to-one interview in Ji'nan, Qingdao, Weifang and Linyi cities of Shangdong province. According to the 1∶1 ratio, local HIV patients with age difference ≤3 years who self-reported MSM were selected as controls. A questionnaire survey was conducted, including data on demographic characteristics, behavior status, CES-D and PSSS. The related factors of the depressive symptoms and characteristics were analyzed. Results: A total of 373 male HIV patients were interviewed, and 39.7%(148/373) were confirmed as MSM after reexamination. The interviewers were: aged (40.3±12.0) years old, 41.9% (62/148) married/cohabiting. 27.0% (40/148) had been tested before HIV diagnosis, 71.6% (106/148) had homosexual partners ≥2 and 55.4% (82/148) had depressive symptom. Multivariate analysis showed that the MSM without HIV testings before diagnosis, had homosexual partners ≥2 before diagnosis, had first homosexual behavior at the age >30 years old and with depressive symptoms were more likely to conceal the true infection route. The incidence of depression among MSM who self-reported HIV infection through heterosexual transmission was related to physical health status social support and occupational stability. Conclusions: Some male HIV persons self-reported as being heterosexually transmitted were really transmitted through homosexual behavior. There were high-risk behaviors such as multiple partners and intersex among this group, and with high incidence of depression. It is necessary to encourage the reexamination program during follow-up and target on psychological and behavioral interventions, continuously.
Male ; Humans ; Adult ; Middle Aged ; Heterosexuality ; Depression/epidemiology* ; HIV Infections/epidemiology* ; Self Report ; Homosexuality, Male ; Cross-Sectional Studies ; Sexual and Gender Minorities

Humans ; Forensic Pathology

Biomechanical Phenomena ; Humans ; Rotator Cuff ; Rotator Cuff Injuries ; Shoulder Joint

ObjectiveTo study the mechanism of renal injury in urogenic sepsis and explore the effect of H2S on the expression of p38 mitogen-activated protein kinase (p38MAPK) and Cysteine protease 3(Caspase3) and apoptosis in renal tissue of urogenic sepsis. Hydrogen sulfide (H2S) has a wide range of biological effects and has a certain protective effect on the kidney. The main objective of this study is to investigate the effect of H2S on the expression of p38 mitogen-activated protein kinase (p38MAPK), cysteine proteinase 3 (Caspase3), and cell apoptosis in renal tissue of urogenic sepsis, and further to study the mechanism of urinary sepsis renal injury.MethodsNew Zealand rabbits (n=40) were divided into five groups Control, Sham, Sepsis, NaHS, and PAG, with eight rabbits in each group. The vital signs, blood routine white blood cell count, neutrophil count, and renal function (Cr, BUN) of five groups of New Zealand rabbits were recorded before the operation, 24 hrs after the operation, 48 hrs after the operation, and 72 hrs after the operation. 72 hrs after the operation, the left kidney tissue was taken for HE staining to observe the changes of cell morphology and structure of the kidney tissue. The apoptotic cells in renal tissue were labeled by Tunel assay (in situ terminal transferase labeling technique). The expression levels of p38MAPK and Caspase3 in renal tissue were tested by ELISA.Results The apoptosis indexes of renal tissue cells in group Control and group Sham were (5.65±2.43)% and (5.57±2.72)%, respectively, with no statistically significant difference (P>0.05). The apoptosis index of rabbit kidney tissue in group Sepsis was (25.26±2.70)%, which was significantly higher than that in group Control and group Sham (P<0.05). The apoptosis index (16.93±2.03)% in group NaHS was significantly lower than that in group Sepsis (P<0.05). The apoptosis index of group PAG (33.09±3.70)% was significantly higher than that of group Sepsis and group NaHS (P<0.05). There was no significant difference between group Control and group Sham (P>0.05). Pairwise comparison between groups Sepsis, NaHS, and PAG showed statistically significant differences (P<0.05). The expression levels of group Sepsis and group PAG were significantly higher than that of group NaHS (P<0.05). The expression level of group PAG was significantly higher than that of group Sepsis (P<0.05).Conclusion H2S can alleviate renal damage caused by urine-derived sepsis, and its mechanism combined with H2S to suppress the expression of p38MAPK and Caspase3 in the renal tissue of urogenous sepsis, thereby reducing renal cell apoptosis Death.

Wnt5a belongs to the large WNT family of cysteine-rich secreted glycoproteins, which is involved in multiple signaling pathways that regulate a variety of cellular processes, including cell motility, proliferation differentiation and so on during development. The regulation and signaling transduction of Wnt5a have been reported to closely relate to inflammatory response, which indicates that Wnt5a plays a critical role in the occurrence and development of inflammatory diseases. In this review, we summarized data on Wnt5a and its signaling pathway, as well as their involvement in inflammatory response. Further comprehensive understanding of the function and relationship between Wnt5a and inflammatory response would help us to develop novel diagnostic and therapeutic strategies for prevention and treatment of inflammatory diseases.
Cell Differentiation ; Cell Movement ; Humans ; Inflammation ; metabolism ; Proto-Oncogene Proteins ; metabolism ; Signal Transduction ; Wnt Proteins ; metabolism ; Wnt-5a Protein

Trichosporon species now ranks as the second most common cause of disseminated yeast infections with a high mortality rate. Breakthrough trichosporonosis in patients receiving echinocandins therapy is being recognized recently. We present a case of breakthrough trichosporonosis with acute viral myocarditis while receiving caspofungin therapy. Trichosporon infection should be considered in patients, who have risk factors for invasive fungal infection and develop unexplained clinical manifestations of infection despite treatment with echinocandins.
Adult ; Antifungal Agents ; therapeutic use ; Echinocandins ; therapeutic use ; Female ; Humans ; Lipopeptides ; Treatment Outcome ; Trichosporonosis ; drug therapy ; microbiology

Objective To determine the effecting factors for diagnosis of binign or malignant in cystic lesions of pancreas(CLP).Methods One hundred twenty-six patients undergoing operations for CLP or suspected CLP in this hospital from January 1984 to June 2008 were reviewed.Patients were divided into two groups according to lesion's histological features after operation.The predictive effect of various preoperative factors on the malignant potential of CLP was evaluated.Results One hundred twenty-six patients underwent operations for suspected pancreatic cystic neoplasms.There were 89 benign and 37 malignant CLPs.The univariate analysis showed that gender, clinical symptoms(jaundice and weight loss), elevated serum CA199, and presence of one or more of three morphologic features such as solid component, nodule or septation were significantly different between benign and malignant cystic neoplasm of pancreas.The multivariate analysis indicated that imaging features and gender were independent predictors of malignancy.Conclusion In patients with suspected pancreatic cystic neoplasms, elevated serum CA199, clinical symptoms(jaundice and weight loss)and presence of suspicious morphologic features on imaging are predictors of malignant potential of CLP.Patients with a high likelihood of a potentially malignant or malignant lesion based on these three factors should undergo operation without additional investigations.

BACKGROUNDMonolayer cell culture models are the traditional culture models used for in vitro research of pancreatic carcinoma chemosensitivity. However, these models neglect the interactions between tumor cells and the impact of the tumor microenvironment. Such tumor cell monolayers poorly mimic the solid tumor microenvironment. The present study aimed to investigate the chemosensitivity characteristics of pancreatic cancer cells in a three-dimensional culture system by analyzing the differences in drug sensitivity between a scattered cell culture model and a multicellular spheroid culture model.

METHODSThree pancreatic cancer cell lines (SW1990, ASPC-1 and PCT-3) were cultured in three-dimensional collagen gels as well as in traditional two-dimensional monolayers. The chemosensitivities of the pancreatic carcinoma cells to 5-fluorouracil (5-FU), gemcitabine, and oxaliplatin in vitro were detected by both the Cell Counting Kit-8 test and the collagen gel droplet-embedded culture drug-sensitivity test.

RESULTSIn the two-dimensional culture model, differences in the chemosensitivities of the cloned pancreatic carcinoma cells and scattered cells existed for some concentrations of 5-FU, gemcitabine and oxaliplatin. In the three-dimensional culture model, there were significant differences in the chemosensitivities of the pancreatic cancer cells between the scattered cells and multicellular spheroids (P < 0.05).

CONCLUSIONPancreatic carcinoma cells exhibit multicellular resistance in three-dimensional cultures.

Antineoplastic Agents ; pharmacology ; Cell Culture Techniques ; methods ; Cell Line, Tumor ; Cell Survival ; drug effects ; Deoxycytidine ; analogs & derivatives ; pharmacology ; Fluorouracil ; pharmacology ; Humans ; Organoplatinum Compounds ; pharmacology ; Pancreatic Neoplasms ; drug therapy ; Spheroids, Cellular ; drug effects

OBJECTIVETo investigate the surgical options for the malignant pancreatic neuroendocrine tumor (PNETs).

METHODSThe clinic data of 32 patients with malignant pancreatic neuroendocrine tumors who were admitted between January 2003 and December 2009 were summarized. There were 19 male and 13 female, aging from 17 to 71 years (mean age, 46.3 years). The general information, postoperative pathology results, surgical options and followup were also collected.

RESULTSThere were 32 cases of malignant PNETs including functional malignant PNETs in 12 cases and nonfunctional malignant PNETs in 20 cases. Thirty-one patients accepted surgical treatments which included local resections in 4 patients, Whipple procedures in 7 patients, distal pancreatectomy in 17 patients and laparotomy in 3 patients. One patient accepted CT guided biopsy. Follow up were lost in two patients after operations. Five patients died in the first year after operations. Twenty-two patients still survived after mean follow up of 28 months (4 - 83 months). The survival rate for this group of patients was 83.5% in the first year, 72.6% in the second year and 54.4% in the fifth year.

CONCLUSIONSMalignant PNETs is the proper indication for surgical treatments. More accumulation of clinical data is needed to evaluate the significance of local resection of malignant PNETs in pancreatic head and cytoreductive surgery.

Adolescent ; Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Neuroendocrine Tumors ; surgery ; Pancreatic Neoplasms ; surgery ; Prognosis ; Retrospective Studies ; Treatment Outcome ; Young Adult