[ ABSTRACT] AIM:To investigate the protective effect of 1, 3-dicyclopentyl-1, 2, 3, 6-tetrahydropyrimidine-4, 5-dicarboxylic acid diethyl ester (ZL-5015) on lethal endotoxin-challenged mice and to explore the underlying mechanism. METHODS:Mouse model of lethal endotoxin challenge and endotoxemia were established by intraperitoneal administration of lipopolysaccharide (LPS) at a dose of 70 mg/kg to the C57BL/6J mice.Mouse peritoneal macrophages stimulated with LPS (10 mg/L) were used as an in vitro inflammatory model.The levels of interleukin-1β( IL-1β) , interleukin-10 ( IL-10) and tumor necrosis factor-α(TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA).Real-time PCR was used to evaluate the mRNA expression of the cytokines.RESULTS:Prophylactic treatment of the mice with ZL-5015 (100 and 200 mg/kg, ig) slightly increased the survival rate, extended the survival time, decreased the serum levels of IL-1βand TNF-α, and increased the serum level of IL-10 in the early stage of endotoxemia as compared with model group.The results of in vitro study demonstrated that treatment of the endotoxin-stimulated mouse peritoneal macrophages with ZL-5015 (10, 20 and 40μmol/L) inhibited the expression of IL-1βand TNF-αat both mRNA and protein levels but promoted the expression of IL-10 at both mRNA and protein levels.CONCLUSION: The tetrahydropyrimidine derivative ZL-5015 shows a moderate anti-endotoxin effect by increasing the survival rate and extending the survival time of the mice challenged by endotoxin, which may result from inhibition of the expression of pro-inflammatory cytokines such as IL-1βand TNF-α, and promotion of the expression of anti-inflammatory cytokine IL-10.

Objective To observe the counteraction of Ganoderma lucidum polysaccharide(GLP) on the inhibition of mice splenocyte proliferation and IL-1? and IL-2 mRNA expression induced by prostaglandin E2(PGE2).Methods Mixed lymphocyte culture reaction(MLR) method was used for the experiment.The lymphocyte proliferation was determined by MTT method,and the levels of IL-1? and IL-2 mRNA expression were evaluated by semi-quantitative reverse transcriptase polymerase chain reaction(RT-PCR).Results After co-culture with PGE2 for 48 hours,the splenocyte proliferation was inhibited to some extent,and the difference was significant when the concentration of PGE2 was over 10?mol/L(P

OBJECTIVETo study the anti-inflammatory and analgesic activities of diethyl 1,3-dicyclohexyl-1,2,3,6-tetrahydropyrimidine-4,5-dicarboxylate (ZL-5010) in vivo and in vitro.

METHODSThe analgesic effect of ZL-5010 was evaluated by acetic acid-induced writhing response in mice, and the anti-inflammatory effects was assessed in mice with xylene-induced ear edema and in rats with carrageenan-induced paw edema. Mouse peritoneal exudate cells activated by bacterial lipopolysaccharides (LPS) were used to evaluate the anti-inflammatory effect of ZL-5010 in vitro. The levels of interleukin-1β (IL -1β) and tumor necrosis factor-α (TNF-α) in the cell culture supernatant were measured using enzyme-linked immunosorbent assay (ELISA).

RESULTSAt the doses of 0.25 and 0.5 mmol/kg, ZL-5010 administered by gavage once daily for 3 days significantly reduced acetic acid-induced writhing frequency and suppressed xylene-induced ear edema in mice, and alleviated paw edema induced by carrageenan in rats (P<0.05). The agent also inhibited the production of the pro-inflammatory cytokines IL-1β and TNF-α by LPS-induced mouse peritoneal exudate cells in vitro, with the statistically significant minimum effective concentrations of 10 and 20 µmol/L, respectively (P<0.05).

CONCLUSIONZL-5010 administered by gavage has anti-inflammatory and analgesic effects in mice and rats, and in mouse peritoneal exudate cell cultures, the agent also inhibits the production of the pro-inflammatory cytokines IL-1β and TNF-α.

Amino Acids, Diamino ; pharmacology ; therapeutic use ; Analgesics ; pharmacology ; therapeutic use ; Animals ; Anti-Inflammatory Agents ; pharmacology ; therapeutic use ; Cyclohexanes ; pharmacology ; therapeutic use ; Female ; Interleukin-1beta ; metabolism ; Male ; Mice ; Mice, Inbred Strains ; Pyrimidines ; pharmacology ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo optimize the experimental model of nitric oxide (NO) production in mouse peritoneal macrophages in response to lipopolysaccharides (LPS) stimulation.

METHODSMouse resident peritoneal macrophages were collected by lavaging the peritoneal cavity of mice with Hank's solution and stimulated with Pseudomonas aeruginosa LPS for NO production. NO concentration in the culture supernatants was measured with Griess Reagent. The influences of cell density, LPS concentration, LPS stimulation duration and culture medium volume on NO production were investigated. Finally, the feasibility of the model was confirmed with specific anti-inflammatory drugs.

RESULTSThe density of macrophages produced the most significant effect on NO production (P<0.001), and optimal results were obtained at the macrophage density of 6×10(6) cells/ml with a volume of 100 µl in each well in 96-well plate. At a LPS concentration below 1 µg/ml, NO production increased proportionally with the increment of LPS concentration (P<0.001), but the increment of NO production declined obviously at LPS concentrations beyond 1 µg/ml, and the peak NO production occurred at a LPS concentration of 10 µg/ml. NO production also increased significantly with the prolongation of LPS stimulation (P<0.05), and the increments were greater within 24-48 h than those in 48-72 h. NO content in the culture supernatant was associated with the medium volume, and the highest level occurred in a system volume of 100 µl. Aspirin (1 mmol/L), dexamethasone (10 µmol/L), and cyclosporin A (10 µmol/L) all significantly inhibited LPS-stimulated production of NO in mouse resident peritoneal macrophages (P<0.001).

CONCLUSIONSMacrophage density, LPS concentration, and the duration of LPS stimulation are the main factors affecting LPS-stimulated NO production in mouse resident peritoneal macrophages. The optimal results can be obtained with a macrophage density of 5×10(6) cells/ml (100 µl per well), LPS concentration of 10 µg/ml, LPS stimulation duration of 24 h or 48 h, and a culture medium volume of 100 to 200 µl.

Animals ; Cells, Cultured ; Female ; Lipopolysaccharides ; pharmacology ; Macrophages, Peritoneal ; drug effects ; secretion ; Male ; Mice ; Mice, Inbred Strains ; Nitric Oxide ; biosynthesis

Objective:To investigate the emergency surgical effect of ruptured intracranial dural arteriovenous fistula (DAVF).Methods:Patients with ruptured intracranial DAVF underwent microsurgery in the Department of Neurosurgery, Nanping First Hospital Affiliated to Fujian Medical University from May 2013 to July 2022 were retrospectively included. The clinical, imaging and follow-up data were collected, and the clinical characteristics, selection of surgical methods and treatment effects of patients were summarized.Results:A total of 8 patients with DAVF were enrolled. Their age ranged from 11 to 60 years (average, 48 years). There were 7 males and 1 female. All 8 patients suffered from intracranial hemorrhage, manifested as headache and vomiting in 2 cases, simple conscious disturbance in 2 cases, conscious disturbance with cerebral hernia in 3 cases, and conscious disturbance with limb paralysis in 1 case. The fistula was located in the anterior fossa in 4 cases (including 2 cases with aneurysms), the middle fossa in 2 cases (including 1 case with moyamoya disease), the transverse sinus in 1 case, and the anterior 1/3 area of the sagittal sinus in 1 case. Cognard classification: 7 patients were type Ⅲ and 1 was type Ⅳ. After admission, all patients underwent emergency craniotomy and microsurgery to remove hematoma. Among them, 4 patients underwent decompressive craniectomy at the same time, 1 patient with moyamoya disease underwent dural turnover and temporalis muscle application at the same time, and 2 patients with aneurysms at the same location were clipped at the same time. Postoperative re-examination of head CT showed that the hematoma was cleared satisfactorily and the midline was no shift in all 8 patients. CT angiography (CTA) showed that the fistula disappeared within 2 weeks. Seven patients were followed up within 1-12 months after operation. CTA or digital subtraction angiography showed no recurrence of DAVF. Two patients with aneurysms did not have residual or recurrent aneurysms. All patients had no new neurological symptoms, and the Glasgow Outcome Scale score in 2 patients increased by 1 compared with that at discharge.Conclusion:Emergency microsurgery is an effective method for the treatment of ruptured intracranial DAVF, especially for patients with special parts or complicated hematoma, cerebral hernia, and other vascular diseases.

Rabies is a zoonotic infectious disease caused by lyssavirus and characterized by central nervous system symptoms. The fatality rate of rabies is almost 100%. About 59 000 cases die of rabies worldwide every year, mainly in Asia and Africa. China is an epidemic country of rabies. Grade II and III exposures are the main types of rabies exposures in China. Standardized post-exposure prophylaxis (PEP) can prevent rabies almost 100%. Human Rabies Vaccine Technical Working Group, National Immunization Advisory Committee and invited experts reached an expert consensus on PEP by referring to the World Health Organization′s position paper on rabies vaccine in 2018 and related research progress in recent.