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Objective: Intracerebral hemorrhage (ICH), the second most common type of stroke, can cause long-lasting disability in the afflicted patients. The study was conducted to examine the patterns of change in endogenous neural stem cells (eNSCs) and in the regenerative microenvironment after ICH, to observe the relationship between the migration of eNSCs and the pattern of change in the polarization state of immune cells in the microenvironment, and provide a research basis for research on clinical nerve repair., Methods: The collagenase injection method was used for modeling. The ICH model was induced in adult female Sprague-Dawley (SD) rats by injecting type VII collagenase (2 U) into the brain tissue of rats. All the experimental rats weighed 280-300 g. In order to simulate the ICU at different time points, including the acute phase (within 1 week), subacute phase (1-3 weeks), and the chronic phase (over 3 weeks), brain tissues were harvested at 3 day post injection (3 DPI), 10 DPI, 20 DPI, and 30 DPI to evaluate the modeling effect. Immunofluorescence staining of the brain tissue sections was performed with DCX antibody to observe the pattern of change in the migration of eNSCs in the brain tissue at different time points. Immunofluorescence staining of brain tissue sections was performed with CD206 antibody and CD86 antibody for respective observation of the pattern of change in pro-inflammatory (M1-type) and anti-inflammatory (M2-type) immune cells in the regenerative microenvironment of the brain tissue after ICM., Results: Spontaneous ICH was successfully induced by injecting type Ⅶ collagenase into the brain tissue of SD rats. The volume of the hematoma formed started to gradually increase at 3 DPI and reached its maximum at 10 DPI. After that, the hematoma was gradually absorbed and was completely absorbed by 30 DPI. Analysis of the pattern of changes in eNSCs in the brain tissue showed that a small number of eNSCs were activated at 3 DPI, but very soon their number started to decrease. By 10 DPI, eNSCs gradually began to increase. A large number of eNSCs migrated to the hemorrhage site at 20 DPI. Then the number of eNSCs decreased significantly at 30 DPI ( P <0.01). Analysis of the immune microenvironment of the brain tissue showed that pro-inflammatory (M1 type) immune cells increased significantly at 10 and 20 DPI ( P <0.01) and decreased at 30 DPI. Anti-inflammatory (M2 type) immune cells began to increase gradually at 3 DPI, decreased significantly at 20 DPI ( P <0.05), and then showed an increase at 30 DPI., Conclusion: After ICH in rats, eNSCs migrating toward the site of ICH first increase and then decrease. The immune microenvironment demonstrates a pattern of change in which inflammation is suppressed at first, then promoted, and finally suppressed again. Inflammation may have a stimulatory effect on the migration of eNSCs, but excessive inflammatory activation has an inhibitory effect on the differentiation and further activation of eNSCs. After ICH, the early stage of repair and protection (10 d) and the subacute phase (20 d) may provide the best opportunities for intervention., Competing Interests: 利益冲突　所有作者均声明不存在利益冲突, (© 2024《四川大学学报（医学版）》编辑部 版权所有Copyright ©2024 Editorial Office of Journal of Sichuan University (Medical Sciences).)

Objective: To observe the effect of penetrative needling of "Baihui" (GV 20) to "Qubin" (GB 7) on neurologic functions and expression of tumor necrosis factor (TNF)-α, interleukin (IL)-6 and toll-like receptor 4 (TLR-4, involving in inflammatory reactions) in the tissue around the local cerebral hematoma in rats with intracerebral hemorrhage (ICH), so as to provide evidence for clinical treatment of ICH., Methods: Fifty-four male Wistar rats were randomly divided into sham control, mo-del and acupuncture groups, and then further divided into three time-point subgroups(1,3,7 days after modeling, n =6/subgroup). The ICH model was established by injection of the rat's autoblood (50 μL) into the putaman region (P:0.2 mm, R:3.5 mm) in a stereotaxic apparatus and confirmed by Berderson's neurologic examination grading system (0-3 points). The neurologic function was assessed by using Longa's scoring (5-points) and footfault asymmetry testing[footfault index=(contra faults-ipsi faults)/total steps in 2 min]. For penetrative needling, an acupuncture needle was inserted into GV 20 and controlled to advance to GB 7 on the affected side and retained for 30 min, once daily. The expression of TNF-α, IL-6 and TLR-4 in the cerebral tissue around the putaman was detected by immunohistochemistry., Results: After penetrative needling stimulation, the increased Longa's score and footfault asymmetry score in ICH rats were significantly decreased on day 1, 3 and 7 after modeling ( P <0.01), suggesting an improvement of neurologic function after the treatment. Immunohistochemical staining outcomes of the cerebral tissue surrounding the autoblood injection site showed that the expression levels of TNF-α, IL-6 and TLR-4 proteins on day 1, 3 and 7 were considerably higher in the model group than in the control group ( P <0.01), and markedly lower in the acupuncture group than in the model group ( P <0.01), suggesting a suppression of the proinflammatory factors and TLR-4 levels around the locus of the brain after needling intervention. A positive correlation existed between the expression levels of TLR-4 and IL-6/TNF-α., Conclusions: Penetrative needling stimulation of GV 20 to GB 7 can reduce the levels of proinflammatory factors TNF-α and IL-6, and TLR-4 in the ICH tissues in rats with cerebral hemorrhage, which may contribute to its effect in improving neurological function.

Objective: To study the changes in humoral immunological function and their clinical significance in cerebral hemorrhage., Methods: Among 115 cases of cerebral hemorrhage, there was low volume bleeding in 39 case, middle volume in 37 cases, and large volume in 39 cases. The ratios of CD19+-CD25+ and CD19+-CD25-B lymphocytes were determined by flow cytometry, and the content of IgA, IgG, IgM, and complement C3 were determined with scattered turbidimetry. The changes in humoral immunological function were compared in patients with different degrees of hemorrhage, and their variation in acute and recovery stages of disease were respectively assessed., Results: The ratios of CD19+-CD25+and CD19+-CD25-B lymphocyte, and the levels of IgA, IgG, IgM, and C3 were higher than those of controls (all P<0.05). The ratios of CD19+-CD25+ and CD19+-CD25- B lymphocyte, and the contents of IgA, IgG,IgM and C3 were increased with the increase in amount of hemorrhage(all P<0.05). The ratios of CD19+-CD25+ and CD19+-CD25-B lymphocytes in recovery stage were lower than those in acute stage (all P<0.05)., Conclusion: There are activations of humoral immunological functions in cerebral hemorrhage. The more abundant in hemorrhage and the severer in disease, the more predominant in the changes in humoral immunological functions.

Objective: To study the influence of beta-endorphin (beta end) on the function of immune system of patients with cerebral hemorrhage at different stages., Methods: Radioimmunal analysis was applied to detect the serum beta-endorphin concentration in the peripheral blood of 28 patients with cerebral hemorrhage, aged 65.5 +/- 13, 28 age-matched patients with cerebral thrombosis, and 28 sex and age-matched normal controls. Mononuclear cells from peripheral blood of these 3 kinds of subjects were cultured and then beta end 10(-8) g/L, beta end 10(-11) g/L, beta end 10(-14) g/L, or beta end 10(-11) g/L + naloxone 10(-5) g/L were added into the media respectively and the MNCs were cultured for more 24 hours (beta end 10(-8) g/L group, beta end 10(-11) g/L group, beta end 10(-14) g/L group, and beta end 10(-11) g/L + Nal group). Another MNCs were cultured without addition of beta end (beta end 0 g/L group). Then the MNCs were collected. RT-PCR was used to detect the expressions of interleukin (IL)-1beta, IL-2, IL-8 and iNOS mRNA in the MNCs., Results: The serum beta-end level of the patients with cerebral hemorrhage at the acute stage was 129 +/- 82 ng/L, significantly lower than that of the normal controls (321 +/- 62 ng/L, P<0.01) and that of the patients with cerebral thrombosis (264 +/- 163 ng/L, P<0.05), but not significantly different from that of the patients with cerebral hemorrhage in the convalescent stage (160 +/- 72 ng/L, P>0.05). The expression of IL-1beta and the expression of IL-2 of the patients with cerebral hemorrhage at the acute stage were significantly lower than those of the patients with cerebral thrombosis and the controls (all P<0.01). The expression of IL-1beta of the patients with cerebral hemorrhage at the convalescent stage were higher than that in the acute stage, however, the difference was not significant (P>0.05). The expression of IL-2 of the patients with cerebral hemorrhage at the convalescent stage was higher than that at the acute stage (P<0.01). The expression of IL-8 and the expression of iNOS of the patients with cerebral hemorrhage at the acute stage were significantly higher than those of the patients of cerebral thrombosis and the controls (both P<0.01). The expression of IL-8 and the expression of iNOS of the patients with cerebral hemorrhage at the convalescent stage were significantly lower than those in the acute stage (both P<0.01). The expressions of IL-1beta, IL-2, IL-8, and iNOS mRNA in the peripheral blood MNCs in vitro in the beta end 10(-8) g/L group and beta end 10(-11) g/L group were significantly higher than those of the beta end 0 g/L group, beta end 10(-11) g/L group, and beta-end 10(-11) g/L + Nal 10(-5)g/L group. The expressions of IL-1beta, IL-2, IL-8, and iNOS mRNA in the peripheral blood MNCs in the beta-end 10(-11) g/L +Nal 10(-5) g/L group were higher than those of the beta end 0 g/L group, however, not significantly., Conclusions: The endogenous beta-endorphin level of cerebral hemorrhage patients is low. The immune system function is up-regulated at the acute stage and then down-regulated. Thereafter the immune system function is invariably low. Exogenous beta-endorphin enhances the IL-1 beta, IL-2, IL-8 and iNOS mRNA expression of peripheral blood MNCS. beta-endorphin receptor antagonist naloxone blocks the positive immunoregulation by beta-endorphin.

The aim of this study is to characterize the pattern of release of soluble vascular cell adhesion molecule-1 (sVCAM-1) in patients with acute cerebral infarction. The level of serum sVCAM-1 was measured serially with two-layer antibody sandwich enzyme-linked immunosorbent assay (ELISA) in 89 patients with acute cerebral infarction, 43 patients with cerebral haemorrhage and 30 normal controls. It is shown that the level of serum sVCAM-1 in patients with cerebral infarction 24 hours after the onset was higher than that in patients with cerebral haemorrhage and normal controls (P < 0.01). The level of serum sVCAM-1 was significantly higher in the patients with large cerebral infarction than that in patients with medium and small cerebral infarctions (P < 0.01). The level of serum sVCAM-1 in patients with cerebral infarction increased from 24 hours to 7 days and decreased from 7 days to 14 days, but it was still higher than that in patients with cerebral haemorrhage and normal controls on the 14th day. The level of serum sVCAM-1 was higher in patients with infection as compared with those without. These findings suggested that sVCAM-1 is closely related to the development of acute cerebral infarction. It is essential to have further study on the change of serumsVCAM-1 in cerebral infarction.

For further understanding the changes in immune function of the patients with hemorrhagic stroke, we investigated the activity of interleukin-2 (IL-2) in the peripheral blood T lymphocytes from 32 patients. The results showed that the levels of IL-2 in the patients were lower than those of the healthy controls, but no significant difference was observed between the cerebral hemorrhage group and subarachnoid hemorrhage group. It suggests the cellular immune function of patients at the acute period is low. We consider that IL-2 might be of use as a kind of therapeutic measure to modulate patients' immune function and treat the diseases.

Fifty-one patients of cerebral hemorrhage were observed on controlled study which were treated with the Zhu-Yu Hua-Tan Decoction (ZYHTD) and Western medicine. The results revealed that ZYHTD could reduce the serum IgG (P < 0.05), and lymphocyte transformation rate (P < 0.05). Meanwhile, the patient's health condition of treated group was better than the control group in the fields of eliminating brain hematoma, evaluating patients condition score in TCM and compromised neurologic function (P < 0.05). It is assumed that there is positive significance in anti-inflammatory effect and promoting the absorption of brain hematoma in the acute phase of cerebral hemorrhage.

The associated antigen of factor VIII in plasma of 124 cases of cerebrovascular disease and 40 normal cases were detected using immune rocket electrophoresis. There were significant differences between the 3 illness groups and the normal group. The results suggest that the associated antigen of factor VIII may be one of the useful markers for forecasting the danger of cerebrovascular disease.