Objective To establish a better method of isolating andculturing ofneural stem cells(NSCs) in neonatal rat brain. Methods Tissue of brain was isolated from neonatal rats. Different medium and culture concentration were used toculture NSCs of neonatal rat. The culture concentration used were 1×10 4, 1×105, 1×106and 1×107/ml respectively. Ingredient of medium was classified into group 1 to 8 respectively according to whether to add 2% B27, epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) as well as the difference in culture concentration. The cells were induced to differentiate asto be confirmed as NSCs, and then were checked by phase contrast microscopy and identified by immunocytochemistry. Results The cells isolated and cultured gathered into neurospheres. The cells were capable of proliferating and maintaining longterm survival in vitro. The cells could be differentiated into neurons and glia.It was to the benefit of the survival of NSCs to add 5% fetal bovine serum(FBS)into the medium at the beginning of the culturing. When 10% FBS was added intothe medium, the neurospheres differentiated quickly. When concentration 1×106/ ml was used, the growth rate of the cells was the highest of all the concentrations. Reasonably higher cell concentration promoted the proliferation of NSCs. It was necessary to add 2% B27, EGF, and bFGF into the medium. The cells had the best growth when 2% B27, 20 ng/ml bFGF and 20 ng/ml EGF were added into the culture medium. EGF and bFGF had cooperative effect. Conclusion A better method of isolating and culturing of NSCs in neonatal rat brain is established and the foundation for future research is laid.
Objective To evaluate the long-term function of the traumaticallydamaged joint after its repair with transplantation of a fresh or a frozen allogenic joint. Methods From March 1977 to September 1993, 13 patients (9 males, 4females; age, 17-55 years) with traumatically-damaged joints underwent transplantation of the fresh or the frozen allogenic joints. Five patients had 5 damagedmetacarpophalangeal joints, 6 patients had 9 damaged interphalangeal joints, and 2 patients had 2 damaged elbow joints. So, the traumatic damage involved 13 patients and 16 joints. All the metacarpophalangeal joints and the interphalangeal joints were injured by machines and the 2 elbow joints were injured by road accidents. The patients were randomly divided into 2 groups: Group A (n=7) andGroup B (n=6). The 7patients with 8 joints in Group A underwent transplantation of fresh allogenic joints; the 6 patients with 8 joints in Group B underwent transplantation of frozen allogenic joints. The allogenic joint transplants were performed in the period from immediately after the injuries to 6 months after the injuries. The motion ranges of the transplanted joints and the X-ray films were examined after operation, and the immunological examination was performed at 8 weeksafter operation. Results The time for synostosis was 5-8 months in Group A, but4-6 months in Group B. In Group A, at 2 years after operation the metacarpophalangeal flexion was 30-40° and the interphalangeal flexion was 20-30°; however,at 6 or 7 years after operation the interphalangeal flexion was only 10-20°. The patients undergoing the transplantation with fresh elbow joints had the elbowflexion of 60° and the elbow extension of 0°, and had the forearm pronation of 30°and the forearm supination of 30°. But in Group B, at 2 years after operation the metacarpophalangeal flexion was 6070° and the interphalangeal flexionwas 40-50°; at 6 or 7 years after operation the interphalangeal flexion was still 40-50°. However, the patients undergoing the transplantation with frozen elbow joints had the elbow flexion of 90° and the elbow extension of 0°, and hadthe forearm pronation of 45° and a forearm supination of 45°. The joint motion ranges, the Xray findings, and the immunological results in the patients undergoing the transplantation of the frozen allogenic joints were significantly better than those in the patients undergoing the transplantation of fresh allogenicjoints. There was a significant difference in the immunological examination between Group A and Group B (IL2, 21.64±3.99；CD4/CD8,3.88±0.82 vs.IL-2,16.63±3.11；CD4/CD8, 2.53±0.23, P＜0.01). Conclusion Repairing the traumatically-damaged joints with frozen allogenic joints is a better method of regaining the contour, movement, and complex motion of the hands.
Objective To investigate the effects of ulinastatin on Treg/Th17 and immune status in patients with severe sepsis.Methods A total of 80 patients with severe sepsis, who were hospitalized in ICU during October 2011 to July 2012, were randomly divided into a routine group and a ulinastatin group. The patients in the ulinastatin group were intravenously administered 30mg ulinastatin three times per day for 5 days in addition to routine bundle treatment. The expression of Treg, Th17 and HLA-DR were detected on the first day in ICU and 5 days after treatment. 20 healthy individuals served as controls. Results Compared with the control group, the severe sepsis group had overexpression of Treg and Th17 ( P lt;0. 01) , higher ratio of Treg/Th17( P lt;0. 01) , and decreased HLA-DR expression of CD14 monocyte ( P lt; 0. 01) . In the severe sepsis patients, ulinastatin injection reduced the abnormal expression of Treg and Th17 ( P lt; 0. 01) , decreased the ratio of Treg/Th17( P lt; 0. 01) , and improved the expression of HLA-DR ( P lt; 0. 01) more effectively compared with the routine treatment. Ulinastatin also lowered 28-day mortality of the patients with sepsis, but the difference between the ulinastatin group and the routine group was not significant. Conclusions In severe sepsis patients, there were abnormal overexpression of Treg and Th17, imbalance of Treg/Th17, and underexpression of HLA-DR which imply an immune suppression. Ulinastatin can decrease the expression of Treg and Th17, inverses the ratio of Treg/Th17, and improve the expression of HLA-DR, so as to improve the prognosis of severe sepsis patients.
In order to investigate pre-and postoperative changes of prostagladin E2 (PGE2) and cellular immune function in patients with gastric cancer (GC), we measured T cell subsets, natural killer cell activity (NKCA) and PGE2 in peripheral blood (PB) of 31 GC patients and 30 controls with APAAP method, LDH release method and radioimmunoassay respectively. The results showed：① a significantly higher levels of PGE2, and a markedly lower levels of CD+4/CD+8 ratio in GC patients before surgery as compared with the controls. The PGE2 had a significant negtive coorelation with CD+4/CD+8 ratio and NKCA respectively.②PGE2 in PB of GC patients gradully declined to normal levels after surgery, moreover PGE2 in tumor tissue was markedly higher than that in normal tissue, implying that the tumor may be the major source of PGE2 in PB. Dynamic determination of PGE2 in PB would be great valuable in evaluation of cellular immune state, to evaluate the effect of surgery and prognosis of patients with gastric cancer.
Objective To investigate the clinical implication on expression of HLA class I in breast cancer tissures.Methods The expression of HLA class I in 271 patients with breast cancer that underwent radical operation was examinedby using immunohistochemically， and the correlation between the expression of HLA class I and clinicalpathological characteristics and prognosis of breast cancer was analyzed. Results The b positive expression of HLA class I in breast cancer tissures was observed in 92 patients （33.9%）， the expressions of HLA class I in 179 patients （66.1%）were downregulation. The expression of HLA class I expression in breast cancer tissures was significantly associated with the axillary lymph node metastasis， TNM stage （P＜0.05）， other lymph node metastasis， and vascular invasion （P＜0.05）. The disease free survival rate of patients with positive expression of HLA class I was higher than that expression downregulation of HLA class I （P＜0.05）. Conclusion The examination of HLA class I expression is useful for the prediction of tumor progression and recurrent risk of breast cancer via the antitumor immune system.
Objective To study the regulative effect of angelica sinensis on cellular immune function in perioperative patients with obstructive jaundice. Methods Fourteen patients with obstructive jaundice were injected with angelica before and after operation for 14 days. The activity of IL-2 and the expression of IL-2R in lymphocytes in peripheral blood were measured, respectively. Results The activity of IL-2 and the expression of IL-2R decreased significantly in patients with obstructive jaundice (P＜0.01). The activity of IL-2 and the expression of IL-2R in peripheral blood lymphocyte increased significantly before and after operations (after treatment using angelica) (P＜0.01), though there was a little decrease after operation but they were still higher than that befor using angelica.Conclusion It maybe useful to use angelica to improve the cellular immune function in patients with obstructive jaundice.
Objective To explore the operative safety of HIV-infected patients with colorectal cancer in different degrees of immunodeficiency. Methods A total of 56 patients, including 26 cases of HIV positive (HIV-positive group) and 28 cases of HIV negative (HIV-negative group), who underwent radical operation for colorectal cancer between January 2012 and December 2015, were enrolled in our study. We divided HIV-positive patients into three groups according to CD4+ T cells count in peripheral venous blood before 1 day (D0) of the surgery (HIV-positive Ⅰgroup with CD4+ T cells count >500/μL, HIV-positive Ⅱgroup with CD 4+ T cells count among 200–500/μL, and HIV-positive Ⅲ group with CD4+ T cells count <200/μL). Non-infective patients were enrolled in HIV-negative group. Leukocyte count, neutrophil percentage, lymphocyte percentage, CD 4+ T cells subsets count, and CD8+ T cells subsets count of the 4 groups in different time points were tested. In addition, we compared postoperative complications, carcinoembryonic antigen (CEA), and postoperative survival rate between the HIV-positive group and the HIV-negative group. Results In 56 cases, there were 26 cases of HIV-positive patients (including 10 cases of HIV-positive Ⅰ group, 8 cases of HIV-positive Ⅱ group and 10 cases of HIV-positive Ⅲ group). Variance results about repeated measurement data showed that, variation of leukocyte count, neutrophil percentage, lymphocyte percentage, and CD8+ T cells count among 4 groups after surgery had no statistical significance (P>0.05), in addition there was no significant on time effect and interactive effect of time and group (P>0.05). CD4+ T cells count in the 4 groups showed a trend from decline to rising with time going, and the time effect had statistical significance (P<0.05). The speed and amplitude of decline and recovery of CD4+ T cells count were different among groups, and the group effect had statistical significance (P<0.05). CEA showed a trend of decline after surgery in both HIV-positive group and HIV-negative group, and the time effect had statistical significance (P<0.05), but the group effect and interactive effect of time and group had no statistical significance (P>0.05). No statistically significant differences in amount of blood loss, duration of surgery, postoperative stay, nor complication rate (including incision infection, pulmonary infection, and opportunistic infections after surgery) were found between the HIV-positive group and the HIV-negative group (P>0.05). The overall survival situation of the HIV-positive group and the HIV-negative group had no statistical significance (P>0.05). Conclusions Radical operation for HIV-infected patients with colorectal cancer has an impact of " first inhibition and recovery” on cellular immunity over a period of time. Incidence of postoperative complications and survival rates are similar in HIV-positive patients and HIV-negative patients. In a word, it’s safe to have radical operation for colorectal cancer in HIV-positive patients under the proper perioperative treatment.