Objective To establish the mice model of immunological tolerance,and investigate the significance of haploidentical allogeneic lymphocytes infusion in induction of graft versus host disease and graft versus tumor in mice.Methods Sixty-four BALB/C female mice were randomly divided into 4 groups with 16 mice in each group.Control group:no special treatment was given after inoculation of tumor cells at the 4th day (CT26 colorectal cancer cell lines with mixture of 1 × 107/mL tumor cells suspension was inoculated to the right subcutaneous axillary of mice) ; Chemotherapy group:chemotherapy was applied at the 7th day after inoculation of tumor cells at the 4th day; DLI group:tumor cells were inoculated at the 4th day,and then haploid donor cells were infused at the 13th,15th and 17th day; Chemotherapy + DLI group:tumor cells were inoculated at the 4th day,chemotherapy was applied at the 7th day,and haploid donor cells were infused at the 13th,15th and 17th day.The pretreatment scheme included haploidentical allogeneic lymphocyte + ring ling amide + haploidentical allogeneic lymphocyte,and the chemotherapy regimen included peritoneal infusion of cyclophosphamide at the 3rd day after inoculation of tumor cells in mice.The time from the first day after vaccination to the day of death of mice and the mass of the tumors were detected to calculate the tumor inhibition rate.The clinical indexes of GVHD were observed,and clinical evaluation was made.The numbers of T lymphocytes in peripheral blood were detected by flow cytometry.Three mice were sacrificed in each group at the 15th day to make the tissue specimens,and they were observed under light microscope after HE staining.All data were analyzed using the analysis of variance or LSD-t test.Results The symptoms of GVHD of mice in the chemotherapy + DLI group were milder than those in other groups.The GVHD scores of the control group,chemotherapy group and the chemotherapy + DLI group were 2.3 ±0.6,1.5 ± 1.1,6.7 ±0.9 and 3.4 ±0.5,respectively,with significant difference between the 4 groups (F =148.68,P ＜ 0.05).The tumor masses of the control group,chemotherapy group,DLI group and the chemotherapy + DLI group were (3.40 ± 0.20) g,(0.80 ± 0.10) g,(2.20 ± 0.20) g and (0.50 ± 0.30) g,respectively,with significant difference between the 4 groups (F =149.17,P ＜ 0.05).The tumor inhibition rates of the control group,chemotherapy group,DLI group and the chemotherapy + DLI group were 0,77％ ± 9％,35％ ± 3％,85％ ± 44％.The levels of CD3 + of the control group,chemotherapy group,DLI group and the chemotherapy + DLI group were 52.3％ ± 2.9％,44.8％ ± 3.1％,62.9％ ± 3.5％,65.9％ ± 3.3％,respectively,with significant difference between the 4 groups (F =28.04,P ＜ 0.05).The levels of CD3 + CD4 + of the control group,chemotherapy group,DLI group and the chemotherapy + DLI group were 32.1％ ± 2.6％,27.1％ ± 1.1％,42.6％ ± 1.8％ and 41.7％ ± 2.4％,respectively,with significant difference between the 4 groups (F =40.29,P ＜ 0.05).The levels of CD3 + CD8 + of the control group,chemotherapy group,DLI group and the chemotherapy + DLI group were 22.7％ ± 2.2％,20.7％ ± 1.8％,26.7％ ± 0.8 ％ and 26.1％ ± 0.7％,respectively,with significant difference between the 4 groups (F =10.74,P ＜ 0.05).The levels of CD3 + CD4 + CD25 + of the control group,chemotherapy group,DLI group and the chemotherapy + DLI group were 8.7％ ±0.6％,6.6％ ±0.6％,11.2％ ±0.4％ and 13.3％ ± 0.7％,respectively,with significant difference between the 4 groups (F =82.88,P ＜ 0.05).Necrosis and bleeding of the tumor tissues were observed in all the 4 groups.Necrosis,shrinking of the tumor cells,inflammatory infiltration were observed in the DLI group and the chemotherapy + DLI group.Proliferation of lymphoid follicles was observed in the chemotherapy + DLI group.The survival time of mice in the control group,chemotherapy group,DLI group,chemotherapy + DLI group were (16.8 ± 2.5) days,(26.3 ± 2.9) days,(23.4 ± 2.5) days and (33.7 ± 4.6) days,respectively,with significant difference between the 4 groups (F =46.45,P ＜ 0.05).Conclusions (1) Pretreatment can induce specific immune tolerance in mice.(2) Haploidentical allogeneic lymphocyte infusion and chemotherapy have synergistic effects,joint application of haploidentical allogeneic lymphocyte infusion and chemotherapy can inhibit the proliferation of tumor cells and prolong the survival time of mice.(3) Chemotherapy can reduce the GVHD of haploidentical allogeneic lymphocyte infusion and enhance the GVT.(4) CD3 + CD4 + CD25 + T lymphocytes play important roles in decreasing GVHD.

OBJECTIVE: To study the effect of melittin on apoptsis and necrosis of osteosarcoma cell line U2 OS in vitro. METHODS: Osteosarcoma cell line U2 OS was treated with melittin. The growth and proliferation was observed by MTT assay and cell counting, and the necrosis was estimated by Trypan blue staining. The cell apoptsis, Fas and Apo2. 7 expression were detected by cytometer. RESULTS: The data showed that melittin could inhibit the proliferation of U2 OS dose-dependently at 16 and 64 mg/L. Cell apoptsis was detected by cytometer, when the cells were treated by 16 mg/L and 32 mg/L of melittin respectively, and the percentages of Fas and Apo2. 7 positive cells were increased. CONCLUSION: Melittin inhibits the proliferation of osterosarcoma cell line through up-regulating Fas expression and inducing apoptsis.

Objective To investigate the clinic signification of newborn hearing screening combined with deafness susceptibility genes screening. Methods 1 440 newborns(3 ~ 5 days after birth) were screened for 8 hot spot hearing loss associated mutations from GJB2, mt12S rRNA and SLC26A4. At the same time, all infants received hearing screening. Those who failed to pass two-step test were referred to further audiological assessment. Results The carrier rate of commonmutations was 1.46% for GJB2 c.235delC, 0.35% for GJB2 c.299-300delAT, 0.42% for mt12S rRNA c.1555A > G, 0.42% for SLC26A4 c.IVS7-2A > G and 0.14% for SLC26A4 c.2168A > G. The total carrier rate was 2.78%. 10 infants were diagnosed as hearing loss in the hearing screening and follow-up audiology assessment (6.94‰) and 5 were diagnosed as severe hearing loss (3.47‰). 32 hearing loss associated mutation carriers passed the hearing screening. Conclusions Genetic screening of newborn hearing screening can be helpful to find out neonates with late-onset and progressive hearing impairment, which were significant for early intervention, regular follow-up and reduction of deafness.

Objective To study the effect of IL-12 on Th1/Th2 cytokine expression level and the role of IL-12 in the shi ft ing of Th1/Th2 immune response against blood-stage Plasmodium b e ighei infection in mice. Methods [WT5”BZ ]BALB/c mice were infected with 5?105 parasitized RBC and received doses of 0.03 or 0.15 ?g/d of IL-12 on the day of infection and daily for 6 days post -in fection. The levels of IFN-? and IL-4 in sera or supernatants of splenic lymp hocyte cultured in vitro were detected by the EL ISA method. Results Compared with spleen cells fro m untreated mice, spleen cells from 0.03 ?g dose of IL-12-treated mice produ ced significantly higher level of Th1-associated cytokine IFN -?, but lower level of Th2-associated cytokine IL-4 in response to PHA or sA g stimulation on day 7 post-infection, whereas spleen cells from 0.15 ?g dose I L-1 2-treated mice produced significantly lower levels of IFN-? and IL-4. The le vel of IFN-? was apparent in the sera of mice treated with 0.03 or 0.15 ?g/d I L-12 on day 3 post-infection and peaked on the day 5 post-infection, but level of I FN-? even was significantly lower in mice treated with 0.15 ?g/d IL-12 comp ara ble to that in control mice on day 7 post-infection. The level of IFN-? was n ot detected in the sera of control mice through 7 days post-infection. [WT5” H Z] Conclusion Appropriate dose of IL-12 regulates the deve lop ment of resistance to P.berghei via a CD 4+ T h1 response, which involves the cytokines IFN-?. [HT5”SS] However, higher doses of IL-12 dramatically inhibite immune pro t ective ability, which may be detrimental to resistance against P .berghei infection. [

Objective To evaluate the results of computer-aid customized acetabular cages for patients with severe defects.Methods Twenty-three patients (8 males and 15 females) with a massive acetabular defects were involved in the present study from January 2005 to September 2014.The average age was 64.2 years (range,46-79 years).According to the American Academy of Orthopaedic Surgeons (AAOS) classification,20 had AAOS type Ⅲ defects and three had AAOS type Ⅳ defects.The customized cages were individualized to each patient's bone defects based on the rapid prototype three-dimensional printed models.The mean follow-up duration was 66.3 months (range,24-120 months).The clinical and radiographic outcomes of all patients were assessed at 6 and 12 weeks after surgery and at once yearly thereafter.Harris hip scores were assessed before surgery and at each follow-up.Postoperative radiographs were evaluated for cage position,migration,and graft incorporation.Complications and reoperations were assessed by chart review.Results The mean Harris hip score improved from 36.2±7.9 (range,20-49) to 81.8± 8.4 (range,60-96),and there is a significant difference between pre-and post-operation (t=23.23,P＜0.001).Individualized custom cages resulted in generally reliable restoration of the hip center.The difference of horizontal distance (between the center of each hip and pubic symphysis) between bilateral sides was-3.0±6.4 mm (range,-19-8 mm).The difference of vertical distance (between the center of each hip and the line connecting the inferior border of the bilateral tear drop) between bilateral sides was 0.4±2.8 mm (range,-4.5-5 mm).No re-revisions had been conducted.None of the cups showed radiographic migration,while one cage was suspected to be loose based on a circumferential 2-mm radiolucent line.Cancellous allografts appeared to be incorporated in 22 of 23 patients.One deep infection and one superficial infection were observed and were treated with irrigation,debridement,and vacuum sealing drainage.One dislocation and one suspected injury of the superior gluteal nerve also were observed and were treated conservatively.Conclusion Individualized customized cages appears to provide stable fixation and improve hip scores at short or mid-term follow-up.

Objective To study the coordination and function of a laparoscopic assistant in laparoscopic pancreaticoduodenectomy (LPD).Methods A retrospective analysis was conducted on 101 patients who underwent LPD at the Department of Hepatobiliary Surgery,Hunan Provincial People's Hospital,from January 2014 to March 2017.The study aimed to study the coordination and function of a laparoscopic assistant.Results LPD was successfully completed in all the 101 patients.There was no conversion to open surgery.The operation time was (326.0 ± 55.6) min,and the resection time was (174.4 ± 42.5) min.The digestive tract reconstruction time was (101.0 ± 21.4) min.The time of pancreaticojejunostomy was (40.5 ± 8.7) min.The time of gastrointestinal anastomosis was:(26.3 ± 5.5) min.The time of biliary anastomosis was (24.4 ± 6.5) min.The intraoperative bleeding was (175.6 ± 41.1) ml.Postoperative pathological data showed that 27 patients (26.7％) had distal common bile duct cancer,23 patients (22.8％)ampullary carcinoma,39 patients (38.6％) duodenal papillary carcinoma,and 12 patients (11.9％) pancreatic ductal adenocarcinoma.The tumor diameter was (2.3 ± 1.3) cm,and the number of resected lymph nodes was (16.7 ±4.2).The number of positive lymph nodes was 1.3 ± 1.1.The length of postoperative hospital stay was 14.8 (8 ～ 29) d.Twenty-three patients developed postoperative pancreatic fistula,including 17 patients (16.8％) with a biochemical fistula,5 patients (5.0％) with a grade B pancreatic fistula,and 1 patient (1.0％) with a grade C pancreatic fistula.There were 2 patients (3.0％) with bile leakage,7 patients (6.9％) with intra-abdominal bleeding,4 patients (4.0％) with delayed gastric emptying,6 patients (5.9％) with abdominal infection,3 patients (3.0％) with pulmonary infection,2 patients (2.0％)with intestinal obstruction,3 patients (3.0％) required a repeated operation,and 1 patient (1.0％) with death in hospital within 30 days after surgery.Conclusions The laparoscopic assistant should have the perspective of "one axis,two sides and four regions" in LPD,and warn the operator to ensure the safety and fluency of the operation by clearly exposing important blood vessels and organs when performing the Kocher incision and when dissecting the key parts such as the dangerous triangle of the uncinate process.During anastomosis,the laparoscopic assistant should appropriately adjust the distance of vision,clearly reveal the surgical field of the anastomotic area,and help the surgeon in improving the precision of the suture and the quality of the anastomosis.

Objective:To evaluate the risk factors of clinical cure and biochemical recurrence (BCR) after radical prostatectomy (RP).Methods:The clinical data of 896 patients who underwent RP at Peking University First Hospital from April 2001 to December 2020 were retrospectively analyzed. Average age was (65.90±6.3) years, median preoperative prostate specific antigen (PSA) was 10.75 (0.36-264.20) ng/ml, median prostate volume was 40.0 (12.0-220.9) ml, median PSA density (PSAD) was 0.27 (0.02-3.42) ng/(ml·g). Clinical staging: 432 cases in T 1c stage, 333 cases in T 2a/bstage, 76 cases in T 2c stage, and 55 cases in ≥T 3 stage. Preoperative Gleason score of biopsy: 193 cases in 3+ 3, 315 cases in 3+ 4, 162 cases in 4+ 3, 226 cases in ≥8. The RP surgery was operated by open or laparoscopic or robot-assisted approach. Clinical cure and BCR were used as the end points for analysis. Clinical cure was defined as a decrease in serum PSA level below 0.03 ng/ml 6 weeks after surgery. BCR was defined as the 2 consecutive serum PSA >0.2ng/ml during the follow-up after RP. Multivariate logistic regression was used to analyze the independent risk factors of clinical cure. The Kaplan-Meier method was used to draw the biochemical recurrence-free survival curve, the log-rank method was used for univariate analysis of BCR, and the Cox regression analysis was used for multivariate analysis. Results:All 896 patients were followed-up for 58 (5-241) months, 678 cases (75.7%) achieved clinical cure. Based on univariate analysis and multivariate analysis, among the preoperative indicators, whether the proportion of positive biopsy needles ≥33% ( P=0.007) and preoperative Gleason score of biopsy ( P=0.041) were independent risk factors of clinical cure. A total of 890 cases were included in the analysis of risk factors of BCR, of whom 172 cases (19.3%) had BCR. The 1-, 5-, and 10-year biochemical recurrence-free survival(BFS)rates were 98.1%, 83.1% and 68.4% respectively. The median BFS has not been reached, and the average BFS was 181 months (95% CI 172-189). The results of univariate and multivariate analysis showed that whether achieved clinical cure ( P=0.001) and postoperative pathological staging ( P<0.001) were independent risk factors of BCR. Conclusions:Whether the proportion of positive biopsy needles≥33% and preoperative Gleason score of biopsy were independent risk factors of clinical cure. Postoperative pathological staging and whether achieved clinical cure may be independent risk factors of BCR.

Humans ; Asthma/drug therapy* ; Biological Therapy