Objective To investigate the action mechanism of IL-35 gene transfection ameliorating cardiac allograft rejection and prolonging allograft survival.Methods pEBI3-L-p35-Fc plasmid was amplified by polymerase chain reaction.In vitro plasmid DNA pEBI3-L-p35-Fc or pSec-L-Fc was,respectively,transfected into HEK293 cells using Lipofectamine 3000.At 48 and 72 h after transfection,IL-35 concentration in culture supernatant of transfected HEK293 cells was detected by ELISA.Balb/c and C57BL/6 splenocytes treated with mitomycin (MMC) served as the stimulators,those not treated with MMC as responders,and they were subjected to one-way mixed lymphocyte culture (MLC).In the presence or absence of IL-35,the percentage of CD4+ CD25+ Tregs was detected by flow cytometry.Abdominal heterotopic heart transplantation model was established by using inbred male Balb/c mice as donors and C57BL/6 as recipients respectively.In experimental group,recipients were intravenously administrated with IL-35 plasmid (50μg) on the day 1 to day 3 post-transplantation.The control mice were treated with normal saline.The IL-35 expression in the blood,CD4+ CD25+ Tregs proportion in the blood and spleen,and the survival and the histopathologic changes of the cardiac grafts were also observed.Results In vitro the transfected HEK293 cells expressed IL-35.IL-35 enhanced the proliferation of CD4+ CD25+ Tregs of MLC in vitro.The median survival time of the cardiac grafts in experimental group (16 days) was significantly longer than in control group (7 days) (P＜0.01).As compared with control group,CD4+ CD25+ Tregs proportion was significantly increased (P＜0.01),CD8+ T cells proportion was decreased (P＜0.01) and the proliferation of lymphocytes and monocytes infiltration was inhibited in the experimental group.Conclusion IL-35 could alleviate cardiac allograft rejection and prolong cardiac allograft survival via the induction of proliferation and differentiation of CD4+ CD25+ Tregs and inhibition of proliferation of CD8 + effector T cells.

Objective To explore the improving effect of L-leucine on memory impairment in plateau and the mecha-nism. Methods After successfully trained in the 8-arm radial maze,50 male Kunming mice were selected and randomly divid-ed into normoxic control group (NC group),model group,and L-leucine (low,medium and high dose) groups.Animals in L-leu-cine groups were intragastrically given 0.473 g?kg-1 ,0.945 g?kg-1 and 1.89 g?kg-1 L-leucine for 7 days and those in NC and model control groups were administered the same volume of purified water for the same period of time.At the 4th day of the treat-ment,the mice in the model control group and L-leucine groups were placed in a large low-pressure and low-oxygen chamber to simulate low-pressure hypoxic environment of the plateau (7 500 m,3 d).The 8-arm radial maze was used to measure the spatial learning and memory ability of mice and dry-wet method to measure the water content of brain tissue.HE staining was employed to observe the cell morphological changes in CA1 region of the hippocampus.The expression levels of mTOR,P70S6K and 4E-BP1 mRNA in the hippocampus were detected by SYBR Green real-time PCR. Results The reference memory error ( RME) ,total error ( TE) ,testing time ( TT) ,and water content of brain tissue were significantly increased,the neuron injury was exacerbated in CA1 region of the hippocampus,and the expression levels of mTOR and P70S6K mRNA were markedly decreased in model control group when compared with those in NC group (P<0.05 or P<0.01).These indexes,however,were significantly improved in L-leu-cine groups,especially in high-dose group. Conclusion L-leucine can improve memory impairment in plateau,and the mecha-nism may involve the activation of mTOR and its downstream substrates (4E-BP1 and P70S6K).

Objective:To explore risk factors of skin pruritus in peritoneal dialysis patients and the effect of individualized care intervention, to provide guidance for clinical practice.Methods:The total of 87 patients with peritoneal dialysis who were followed-up with pruritus in the Beijing Anzhen Hospital from January 2017 to June 2020 were selected. The Visual Analogue Scale (VAS) was used to evaluate the degree of pruritus, and the patients were divided into two groups: mild-to-moderate skin pruritus group (VAS≤6 points) and severe skin pruritus group (VAS>6 points). The risk factors of severe skin pruritus were analyzed by single factor and multivariate Logistic regression. The improvement of skin pruritus after 3 months of individualized nursing intervention was observed.Results:Among the 87 patients, the mild-to-moderate skin pruritus group and the severe skin pruritus group accounted for 64.4%(56/87) and 35.6%(31/87), respectively. Single factor analysis showed that the age, prevalence of diabetes, serum albumin, serum phosphorus, intact parathyroid hormone and C-reactive protein levels were (61.8 ± 11.5) years old, 33.3%(19/56), (36.3 ± 5.3) g/L, (1.6 ± 0.5) mmol/L, 328.4(144.9, 494.5) ng/L, 2.8(0.6, 8.3) ng/L in the mild-to-moderate skin pruritus group, and (67.0 ± 9.2) years old, 61.1%(19/31), (33.9 ± 4.8) g/L, (1.9 ± 0.3) mmol/L, 397.0(300.0,758.6) ng/L, 7.2(2.6, 17.2) mg/L in the severe skin pruritus group, the differences were significant between the two groups ( t values were -2.17, 2.14, -2.32, Z values were -2.28, -2.90, χ 2 value was 6.07, P<0.05). Logistic multivariate regression analysis showed that low albumin, high blood phosphorus and high C-reactive protein were independent risk factors for severe skin pruritus in peritoneal dialysis patients ( P<0.05). After 3 months of individualized care,18.4% (16/87) patients had complete remission,19.5% (17/87) patients significantly relieved, 55.2% (48/87) relieved, 6.9% (6/87) were ineffective, and the total response rate was 93.1%(81/87). Conclusions:More than one-third of peritoneal dialysis patients with pruritus are severe. Lower serum albumin, higher serum phosphorus and higher C-reactive protein are independent risk factors for severe pruritus in peritoneal dialysis patients. Individualized care can effectively improve pruritus in peritoneal dialysis patients.

OBJECTIVETo investigate the feature of histomorphology and biology of uterine bizarre leiomyoma from the clinical, pathological features as well as the immunohistochemical expression.

METHODSTotally 25 cases of leiomyomas were studied. Among them, immuno-histochemical staining (SP and ABC methods) for smooth muscle actin (SMA), proliferative cell nucleus antigen (PCNA), estrogen receptors (ER), and progesterone receptors (PR) were performed in 20 cases. The clinical features and follow-up records were analyzed.

RESULTSThe main clinical findings were irregular vaginal bleeding, pain and pelvic tumor. One case was with immense amount of ascitis and the other two were with pregnancy. All the cases had no history of taking pregestine. Light microscopy showed that part of all the cell nuclei were bizarre, accompanying with double or multiple nuclei in which rather big and reddish staining inclusion bodies were obtained (D = 7 - 26 micro m), and the mitotic figures were 0 - 2/10 HPF. Among 20 cases with immunohistochemical staining, markers indicating muscle cells in origin were positive in the bizarre cells, 15 of which (75%) with negative or weak positive PCNA, and 18 of which (90%) with negative ER. There were significant difference between the studied and control groups (P = 0.027, P < 0.005 respectively) and in addition, PR was positive in both these two groups. A majority of the nucleus inclusion bodies was SMA positive. Follow-up records demonstrated so far no recurrence cases obtained.

CONCLUSIONSUterine atypical leiomyoma belongs to benign tumor, although its shape is bizarre. In this group, the morphology changes are correlated with pregnancy, but not with pregestine. The expression of immunohistochemistry shows certain features, and is important to identify uterine bizarre leiomyoma with leiomyosarcoma and STUMP in pathological diagnosis.

Adult ; Female ; Humans ; Immunohistochemistry ; Leiomyoma ; metabolism ; pathology ; Middle Aged ; Pregnancy ; Uterine Neoplasms ; metabolism ; pathology

Objective: To observe assessment effects of thromboelastogram (TEG) on patients with coronary heart disease (CHD) complicated hypertension.Methods: A total of 120 CHD patients were selected from our hospital.According to complicated with hypertension or not, they were divided into pure CHD group (n=58) and CHD + hypertension group (n=62).TEG indexes were compared between two groups.Results: Compared with pure CHD group, there were significant reductions in blood clot formation duration [K: (2.53±0.72)min vs.(1.82±0.64)min], coagulation reaction duration [R: (8.66±1.86)min vs.(7.18±1.85)min], arachidonic acid pathway-induced platelet activity [AA: (57.36±16.91)% vs.(46.73±20.73)%], and significant rise in maximum amplitude after clot formation [MA: (57.31±7.75)mm vs.(64.36±7.85)mm] and included angle value between the tangent from the blood clot forming point to the maximum curve radian of the chart and the horizontal line [Angle: (53.26±7.78) vs.(64.38±7.85)] in CHD + hypertension group, P<0.01 all.Spearman correlation analysis indicated that blood pressure level was significantly positive correlated with Angle and MA (r=0.607, 0.405, P<0.01 both), and significantly inversely correlated with R and K (r=-0.256,-0.541, P<0.01 both) in CHD + hypertension patients.Conclusion:Thrombosis possesses higher risk for CHD + hypertension patients, which is easier to cause acute cardiovascular events.Therefore, attention should be paid to coagulation function monitoring in order to prevent adverse cardiac events in these patients.

Objective To elucidate the clinicopathological characteristic, differential diagnosis, treatment and prognosis of systemic amyloidosis. Methods An inpatient diagnosed as systemic amyloidosis was analyzed for clinical and pathological features as well as laboratory findings. The related literature was reviewed. Results The patient was confirmed to have amyloidosis of the muscle. Muscle involvement was the most prominent and first manifestation, and the patient had widespread visceral involvements, which included cardiovascular system, kidney, respiratory as well as gastrointestinal tracts and tongue. The biopsy of the muscle, mucosa of stomach and intestine, and cutaneous tissue revealed amyloid material deposited in the skeletal and smooth muscle as well as vessel walls. Conclusion Amyloid myopathy is a rare manifestation in systemic amyloidosis. Skeletal muscle weakness and stiffening may be an important clue to the diagnosis of systemic amyloidosis.

Objective: To evaluate the anti-tumor and side effects of activated-HLA haploidentical peripheral blood tem cells (haplo PBSCs) in the treatment of advanced refractory solid tumor patients. Methods: Forty-two patients with advanced refractory tumor, who were diagnosed in our hospital from Oct. 2004 to Oct. 2007, were enrolled in this study (all patients signed informed consent), including 12 with ovarian cancer, 9 with renal cancer, 8 with lung cancer, 8 with breast cancer, 2 with colon cancer, 2 with gastric cancer, and 1 with melanoma. The donors were healthy direct relatives of the patients; the donors' haplo-PBSCs were mobilized, collected, and activated by rhIL-2 in vitro. The clinical efficacy and side effects of haplo-PBSCs therapy were assessed by CT/PET-CT scanning, RESIST standard, KPS score, and clinical response rates, etc. Results: All 42 patients received one episode of haplo-PBSCs treatment. The progression-free survivals (PFS) were 6 months and the clinical beneficial rate (CR+PR+SD) was 73.8%. The beneficial rate of life quality was 76.2% and the KPS increased by 20 (0-30) points on average after haplo-PBSCs treatment. The patients with KIR unmatched in GVH direction had better outcomes than those with KIR matched or KIR unmatched in HVG direction (P<0.05), and the clinical beneficial rate, PFS and total beneficial rate were 94.1% vs 60.0%, (13.4±1.3) vs (8.0±0.9) months, and 89.5% vs 65.2%, respectively (all P<0.05). The donor/recipient relation as the mother/child had a better outcome than that as the father/child (P<0.05). Patients with renal cancer or ovarian cancer had better outcomes than those with other cancers, with clinical beneficial rates being 90.0% and 81.8%, respectively. Conclusion: Activated haplo-PBSCs therapy can induce non-specific anti-tumor effect, and improve the clinical symptom and life quality of advanced tumor patients.

Objective To study the characteristics of auditory brainstem response by chirp and click stimuli in normal hearing children.Methods A total of 97 children (194 ears)with normal hearing were recruited,including 50 male (100 ears),and 47 female (94 ears).The CE-chirp and the click acoustic stimulation were used to,and the ABR waveforms and latencies were obtained.ResuIts The chirp-ABR Ⅰ wave recognition rate (36%)was significantly lower than that of click-ABR (100%).At 80 dB nHL,chirp-ABR V wave latency was shorter than click-ABR,the difference was statistically significant (P< 0.05 ).The chirp and click ABR V wave response thresholds showed no significant difference (P>0.05).At 80 dB nHL,no statistical significance of two kinds of a-coustic stimulation of ABR V wave amplitude difference (P>0.05).At 50 dB nHL,chirp-ABR significantly in-creased than click-ABR V wave amplitude with statistically significant differences (P<0.05).At 80,and 50 dB nHL,chirp-ABR showed no differences in the amplitude of V wave (P>0.05 ).ConcIusion The chirp acoustic stimulation produced the larger response amplitude,and was faster,demonstrating the noticeable advantages to tes-ting children.

Objective To investigate the effects of sirolimus and 3-methyladenine (3-MA) on the autophagy in,as well as matrix matalloproteinase (MMP)-1 and MMP-3 secretion by,human skin fibroblasts (HSFs).Methods HSFs were isolated from the circumcised foreskin of a healthy male,and subjected to primary culture.After 3 to 10 passages,HSFs were incubated with sirolimus of 20,50,100,250 nmol/L and 3-MA of 0.5,2.0,5.0,10.0 mmol/L respectively for 4 hours followed by the observation of autophagy and detection of MMP-1 and MMP-3 levels in the supernatant by enzyme linked immunosorbent assay (ELISA).Those HSFs remaining untreated or treated with dimethyl sulfoxide served as the control.Results The percentage of autophagy-positive cells was 59.075％ ± 6.884％,76.350％ ± 5.226％,85.063％ ± 6.002％,86.288％ ± 5.558％ and 96.825％ ± 1.500％ respectively in HSFs treated with sirolimus of 0,20,50,100 and 250 nmol/L; significant differences were observed between the 5 groups (P ＜ 0.01 ) but not between the cells treated with sirolimus of 50 and 100 nmol/L (P ＞ 0.05).After being treated with 3-MA of 0,0.5,2.0,5.0 and 10.0 mmol/L,the percentage of autophagy-positive cells in HSFs was 63.037％ ± 5.876％,34.425％ ± 5.183％,19.700％ ± 3.028％,12.900％ ± 3.334％ and 7.775％ ± 2.293％ respectively with a significant difference between these groups (all P ＜ 0.01 ).Elevated levels of MMP-1 and MMP-3 were observed in the supernatant of HSFs treated with sirolimus of 250 nmol/L and 3-MA of 10.0 mmol/L (all P ＜ 0.05).Conclusions The autophagy in HSFs can be upregulated by sirolimus,but downregulated by 3-MA.For the secretion of MMPs by HSFs,3-MA and high concentrations of sirolimus exert a promotive effect,and the effect of 3-MA is in a concentration-related manner,but the influences of sirolimus at lower concentrations remain unclear.

Objective To observe the changes of autophagy in human skin fibroblast (HSF) model for photoaging. Methods HSF model for photoaging was established through repeated exposure to ultraviolet B (UVB). Those HSFs receiving no irradiation served as the control. The degree of aging was evaluated by p-galactosidase assay, and autophagy level was detected. Results After repeated exposure to UVB, most pho-toaged HSFs were deformed and distorted, and some of them even died. The percentage of P-galactosidase-positive cells was 50.60% ± 5.04% and 14.58% ± 2.69%, respectively in photoaged and control HSFs (P< 0.01). Significant difference was also observed in the proportion of cytophagosome-positive cells between photoaged and control HSFs (14.91% ± 4.59% vs 68.45% ± 8.25%, P < 0.01). Conclusion The HSF model for photoaging shows obviously abnormal appearance and stagnant growth with increased degree of senescence and decreased autophagy compared with normal control HSFs.