Tong Luo Jiu Nao injection, a traditional Chinese medicinal preparation, inhibits MIP-1ß expression in brain microvascular endothelial cells injured by oxygen-glucose deprivation.

ETHNOPHARMACOLOGICAL RELEVANCE: Tong Luo Jiu Nao injection (TLJN), a modern Chinese formula based on Traditional Chinese Medicine theory, has been used to treat ischemic stroke and vascular dementia. TLJN belongs to the ethnopharmacological family of medicines. AIM OF THE STUDY: To investigate the protective effect of TLJN on oxygen-glucose deprivation (OGD) induced-injury of brain microvascular endothelial cells (BMECs). MATERIALS AND METHODS: The model of OGD was established in the primarily cultured BMECs. TLJN was added to the OGD-injured BMECs for 6h. A series of assays were used to detect the effects of TLJN on: (i) MIP-1ß content in BMECs conditioned media (CM) by ELISA; (ii) MIP-1ß protein expression in BMECs by western blotting and immunocytochemistry; (iii) the expression of CCR5, receptor of MIP-1ß, in BMECs by western blotting; (iv) the proliferative activity of microglial cells via the Cell Counting Kit-8 (CCK-8). RESULTS: Our results showed that the OGD-injured BMECs presented with large amounts of secretion and expression of MIP-1ß and up-regulation of CCR5. Also, the CM of OGD-injured BMECs remarkably increased the proliferative activity of microglial cells. The TLJN-treated BMECs exhibited a reduction of MIP-1ß content in BMECs-CM and a down-regulation of MIP-1ß and CCR5 expression. In addition, an inhibitory effect of CM of OGD-injured plus TLJN injection-treated BMECs on microglial proliferation was also found. CONCLUSION: TLJN reduced the expression of MIP-1ß and CCR5 in OGD-injured BMECs, and the CM of OGD-injured plus TLJN injection-treated BMECs inhibited the proliferative activity of microglial cells, suggesting the therapeutic potential of TLJN on ischemic cerebral vascular disease.

Impact of paracrine signals from brain microvascular endothelial cells on microglial proliferation and migration.

Neuronal survival can be influenced by activated microglia, but limited evidence exists on the effects of paracrine signaling from brain microvascular endothelial cells (BMECs) on microglial action. Therefore, we examined the effects of normal BMECs conditioned medium (BMECs-CM) on activated microglia induced by pro-inflammatory cytokine macrophage inflammatory protein-1beta (MIP-1ß), a chemokine that released from ischemic BMECs and has been proved to stimulate microglial proliferation. Our results showed that BMECs-CM inhibited the proliferation and transmigration of microglia induced by MIP-1ß. Moreover, BMECs-CM significantly suppressed the expression of the MIP-1ß receptor, CCR5, and the phosphorylation of p38 and JNK (P<0.05). These findings suggest that BMECs-CM could inhibit MIP-1ß-induced microglial activation. Future therapeutic strategies that prioritize the early recovery of BMECs could be beneficial for microglial inhibition and further increase neuronal survival.

[Polymerase region mutations and hepatitis B virus genotypes in chronic hepatitis B patients with poor response to lamivudine].

OBJECTIVE: To investigate the mutations in Polymerase region and hepatitis B virus (HBV) genotypes in chronic hepatitis B patients with poor response to Lamivudine treatment. METHODS: 631 chronic hepatitis B patients with poor response to Lamivudine were recruited in this study. Real-time PCR and DNA sequencing were used to determine HBV genotypes; direct sequencing was performed to detect mutations, and real-time PCR was used to quantify HBV DNA load. Mutations in polymerase region were investigated in different HBV genotypes. RESULTS: 272 patients were infected with HBV of genotype B, and 359 patients were infected with HBV of genotype C. The mean age of patients infected with HBV of genotype C (39.1+/-11.4 years old) were significant higher than that of patients infected with HBV of genotype B (33.7+/-9.7 years old) (t = -6.55, P less than 0.01). The patients infected with HBV of genotype C had relatively higher HBV DNA load [(5.96+/-1.22) log10 copies/ml] than the patients infected with HBV of genotype B [(5.58+/-1.21) log10 copies/ml] (t = -2.01, P less than 0.05). The overall incidence rate of A181V/T mutation in genotype C (5.3%) was significantly higher than that in genotype B (0.4%) (x2=12.23, P less than 0.01), but the incidence rate of M204I/V, L180M, T184A/G/I/S, S202G/I and V173L mutations was not significantly different between genotype B and C (each P more than 0.05). M204I mutation in genotype B (20.6%) was more frequent than that in genotype C (13.9%) (x2=4.91, P less than 0.05). The Lamivudine resistance mutations were not significantly different between genotype B and genotype C (x2 = 0.00, P more than 0.05). CONCLUSIONS: The incidence rate of lamivudine resistance mutation is not significantly different between genotype B and genotype C, but patients infected with HBV of genotype C have higher HBV DNA load than patients infected with HBV of genotype B.

[IgM to recombinant antigen E2 of HEV envelope protein is a sensitive and specific serological marker in diagnosis of acute hepatitis E].

OBJECTIVES: To evaluate the sensitivity and specificity of IgM to recombinant antigen E2 of HEV envelope protein in the diagnosis of acute sporadic hepatitis E. METHODS: anti-E2 IgM was detected in sera samples from 176 cases of acute sporadic hepatitis E and 191 cases of acute non A-E hepatitis by ELISA and was compared with HEV IgM detected by some domestic and Genelabs (GL) kits. HEV RNA was also detected in sera positive for anti-E2 IgM. Logistic regression was used to analyze factors associated with the detection of anti-E2 IgM and HEV RNA. RESULTS: Anti-E2 IgM was found in 68.75% of the serum samples from the 176 patients of acute hepatitis E and the positive rate of HEV IgM detection by domestic kits was 56.25% (chi2 IgM = 6.49, P < 0.05). There were 37 (19.37%) anti-E2 IgM positive cases in those 191 sera of the acute non A-E hepatitis, out of which 11 cases were also detected as positive by the GL kit. Of the 158 anti-E2 IgM positive sera, HEV RNA was found in 81 (51.27%), among which 57.02% was positive in acute hepatitis E and 32.43% in acute non A-E hepatitis. No HEV RNA was found in the anti-E2 IgM negative sera from the cases of acute hepatitis E. By logistic regression analysis, no variance relative to the detection of anti-E2 IgM was found with the time interval from onset to hospitalization, age, levels of bilirubin, ALT and AST of the serum. Only the level of serum ALT was found being significantly associated with the detection of HEV RNA (P = 0.024). CONCLUSIONS: (1) anti-E2 IgM is a sensitive and specific serological maker for diagnosing an acute infection of HEV. (2) HEV is still the pathogen of some cases diagnosed clinically as non-A-E hepatitis. (3) Persistent HEV viremia is possibly an important factor influencing the severity of acute hepatitis E.