Anesthesia, General ; adverse effects ; Child, Preschool ; Humans ; Male ; Malignant Hyperthermia ; diagnosis ; etiology

Female ; Humans ; Infant ; Meningitis, Bacterial ; complications ; Salmonella Infections ; complications ; Salmonella typhi ; isolation & purification ; Sepsis ; complications

Objective To investigate the effects of shRNA targeting a disintegrin and metalloproteinases 17(ADAM17)on the proliferation of human breast cancer MCF-7 cells in hypoxia environment.Methods The four specific ADAM17-shRNA se-quences aiming at ADAM17 were designed,transfected into MCF-7 cells by electroporation,and cultured in hypoxia environment. The experiment was divided into the control group (blank phosphate buffer solution,PBS),nonsense sequence group (transfected with ADAM17-shNC)and shRNA transfection group (transfected with ADAM17-shRNA,the highest silencing efficiency of shR-NA was selected for following experiments).Real-time PCR was used to detect the expression of ADAM17 mRNA.The prolifera-tion ability and cell cycle change of MCF-7 cells were detected by iCELLigence and flow cytometry (FCM),respectively.Results Compared with control group and nonsense sequence group,the four ADAM17-shRNA transfection groups all had the silence effect on ADAM17 gene expression (P <0.05 ),the difference was statistically significant(P <0.05 ),particularly shRNA1219 had the highest inhibitory rate (F =5.11,P <0.01 ).The cellular proliferation ability and cell growth speed in the shRNA transfection group were significantly decreased compared with the control group and nonsense sequence group (P <0.05).Most cells of shRNA transfection group remained in the G0/G1 phase (73.35 ± 2.45 ),which in the control group and nonsense sequence group was (62.56±2.35)and (62.68 ±1.20)respectively,the difference was statistically significant(P <0.05).The cell cycle progression was significantly delayed.Conclusion ADAM17-shRNA inhibits the proliferation of MCF-7 cells under hypoxic environment.

Child, Preschool ; Female ; Hand, Foot and Mouth Disease ; complications ; diagnosis ; therapy ; Humans ; Infant ; Male ; Pulmonary Edema ; etiology ; Radiography, Thoracic

ObjectiveTo investigate the effects of leukocyte-depleted allogeneic blood transfusion on perioperafive cellular immunity in children.MethodsThree hundred and fifty-nine ASA Ⅰ or Ⅱ children aged 3 month-14 yr,weighing 5-74 kg requiring allogeneic blood transfusion during operation were randomly divided into 2 groups:control group (group C,n =163) and leukocyte depletion group (group D,n =196).In group D allogeneic blood was filtered with a leukocyte filter before being transfused during operation.Blood samples were collected from peripheral vein before blood transfusion,and at 2 and 6 d after blood transfusion for determination of levels of CD3+,CD4+,CD8+,and CD56+ by flow cytometry.CD4+/CD8+ ratio was calculated.The volume of allogeneic blood transfused during operation,the duration of operation,postoperative drainage,antibiotics administration and hospital stay and incidence of postoperative infection were recorded.ResultsThe levels of CD3+,CD4+,CD56+ and CD4+/CD8+ ratio were significantly increased at 6 d after blood transfusion while the duration of postoperative drainage,postoperative antibiotics administration and hospital stay and incidence of postoperative infection were significantly decreased in group D compared with group C.ConclusionLeukocyte-depleted allogeneic blood transfusion is helpful in improving the postoperative cellular immunity in children.

OBJECTIVE To determine the functional role of hydrogen sulfide (H2S) in protecting against mitochondrial dysfunction in heart failure through the inhibition of Ca2 +/calmodulin-dependent protein kinaseⅡ (CaMKⅡ) using wild type and CSE knockout mouse models. METHODS Continuous subcutaneous injection isoprenaline (7.5 mg·kg-1·d-1), once a day for 4 weeks to induce heart failure in Male C57BL/6 (6-8 weeks old) mice and CSE-/- mice. 150 μmol·L-1 H2O2 was used to induce oxidative stress in H9c2 cells. Echocardiograph was used to detect cardiac parameters. H&E stain and Masson stain was to observation histopathological changes. Western blot was used to detect protein expression and activity. The siRNA was used to silence protein expression. HPLC was used to detect H2S level. Biotin assay was used to detect the level of S- sulfhydration protein. RESULTS Treatment with S-propyl-L-cysteine (SPRC) or sodium hydrosulfide (NaHS), modulators of blood H2S levels, attenuated the development of heart failure in animals, reduced lipid peroxidation, and preserved mitochondrial function. The inhibition CaMKⅡ phosphorylation by SPRC and NaHS as demonstrated using both in vivo and in vitro models corresponded with the cardioprotective effects of these compounds. Interestingly, CaMKⅡ activity was found to be elevated in CSE-/- mice as compared to wild type animals and the phosphorylation status of CaMKⅡ appeared to relate to the severity of heart failure. Importantly, in wild type mice SPRC was found to promote S-sulfhydration of CaMKII leading to reduced activity of this protein however, in CSE-/- mice S-sulfhydration was abolished following SPRC treatment. CONCLUSION A novel mechanism depicting a role of S-sulfhydration in the regulation of CaMKⅡ is presented. SPRC mediated S-sulfhydration of CaMKII was found to inhibit CaMKⅡ activity and to preserve cardiovascular homeostasis.

OBJECTIVE To determine the functional role of hydrogen sulfide (H2S) in protecting against mitochondrial dysfunction in heart failure through the inhibition of Ca2 +/calmodulin-dependent protein kinaseⅡ (CaMKⅡ) using wild type and CSE knockout mouse models. METHODS Continuous subcutaneous injection isoprenaline (7.5 mg·kg-1 per day), once a day for 4 weeks to induce heart failure in male C57BL/6 (6-8 weeks old) mice and CSE-/- mice. 150 μmol·L-1 H2O2 was used to induce oxidative stress in H9c2 cells. Echocardiograph was used to detect cardiac parameters. H&E stain and Masson stain was to observation histopathological changes. Western blot was used to detect protein expression and activity. The siRNA was used to silence protein expression. HPLC was used to detect H2S level. Biotin assay was used to detect the level of S-sulfhydration protein. RESULTS Treatment with S-propyl-L-cysteine (SPRC) or sodium hydrosulfide (NaHS), modulators of blood H2S levels, attenuated the development of heart failure in animals, reduced lipid peroxidation, and preserved mitochondrial function. The inhibition CaMKⅡ phosphorylation by SPRC and NaHS as demonstrated using both in vivo and in vitro models corresponded with the cardioprotective effects of these compounds. Interestingly, CaMKⅡ activity was found to be elevated in CSE-/- mice as compared to wild type animals and the phosphorylation status of CaMK Ⅱ appeared to relate to the severity of heart failure. Importantly, in wild type mice SPRC was found to promote S-sulfhydration of CaMKⅡ leading to reduced activity of this protein however, in CSE-/- mice S-sulfhydration was abolished following SPRC treatment. CONCLUSION A novel mechanism depicting a role of S-sulfhydration in the regulation of CaMKⅡ is presented. SPRC mediated S-sulfhydration of CaMKⅡ was found to inhibit CaMKⅡ activity and to preserve cardiovascular homeostasis.

Objective To validate a new standardized training program for urological surgeons to improve their laparoscopic surgical skills. Methods The laparoscopic surgical training program was carried out by using the traditional mechanical simulators and animal models. Thirty-three trainees participated the urological laparoscopic surgical training program. Initially, the novices were assigned to practise basic laparoscopic skills step by step on the simulator with fixed trocar positions. After a period of basic training, they were allowed to practise on animal models for some particular proce-dures. Results All trainees (33/33, 100.0%) participated were able to perform all basic techniques skillfully and completed laparoscopic anastomosis accurately after the training. The time required for performing the partial nephrectomy, dismembered pyeloplasty and ureteral reimplantation on animal models declined from 64.0±18.4, 127.54±17.5 and 75.84±11.6 min at the beginning to 30.94±3.8, 65.2±7. 5 and 37.7±7.2 min after practicing these procedures 8 times (P<0.01). They could un-derstand the crucial procedures of the laparoseopic surgeries after 6 to 8 special trainings on animal models. Fifteen trainees (15/33, 45.5%) had started to carry out laparoscopic surgeries after-finish- ing the training program. Conclusions Our program enables the participants to improve their techniques in complicated laparoscopic surgeries. The challenging parts of reconstructive laparoscopic surgeries such as laparoscopic dismembered pyeloplasty can be taught by using animal models. This program could be incorpo-rated easily by all urological departments developing a laparoscopie surgical training program.

Objective To investigate the clinical feature , pathologic characteristics and prognosis of urothelial carcinoma of bladder with squamous differentiation . Methods From Jan.2010 to Jun.2013, the pathological and clinical data of 96 cases of urothelial carcinoma of bladder with or without squamous dif-ferentiation were compared .Of the group with squamous differentiation , there were 39 males and 9 females with a median age of 70 (29 to 87) years.44 cases presented with painless gross hematuria .4 cases presen-ted with finding of bladder tumors in annual physical examination .TURBT, partial cystectomy and radical cystectomy were performed in 25, 8 and 13 cases, respectively.In addition, one case was only underwent bi-lateral ureteral skin gastrostomy .The last one only performed cystoscopy .In accordance with sex , age, path-ological stage and classification and surgical approach , 48 controls were selected .For the other group , there were 40 males and 8 females with a median age of 68 (39 to 86) years.45 cases presented with painless gross hematuria.3 cases presented with finding bladder tumors by annual physical examination .TURBT, par-tial cystectomy and radical cystectomy were performed in 28, 7 and 13 cases, respectively.All patients with retaining bladder had postoperative intravesical instillation for one year .Some patients with or without bladder performed 3-6 cycles chemotherapy with the GC protocol . Results In squamous differentiation group , there were 1 (2.1%) pTa, 25 (52.1%) pT1, 17 (35.4%) pT2, 4 (8.3%) pT3 and 1 (2.1%) pT4 tumors. Whereas, 1 (2.1%) pTa, 28 (58.3%) pT1, 16 (33.3%) pT2, 2 (4.2%) pT3 and 1 (2.1%)pT4 tumors were selected in the control group .There were 2 (4.2%) cases with low grade and 46 (95.8%) cases with high grade carcinomain in both groups .Patients were followed up with a mean duration of 16 and 12 months in squamous differentiation and control group , respectively .In squamous differentiation group , eight recur-rences were recorded with a mean follow-up of 12 months.Of the 3 died patients, only one died from bladder cancer.In control group, seven recurrences were recorded with a mean follow-up of 22 months, and no pa-tient died.For patients with TURBT, 3 year recurrence rate of patients with squamous differentiation was 49.5%, while the control was 34.8%.The difference was statistically significant (P<0.05). Conclusions Urothelial carcinoma of bladder with squamous differentiation is at a high level of malignant and recurrence . The rate of myometrial invasion with squamous differentiation is higher than pure urothelial bladder cancer . Patients with squamous differentiation should be closely followed up .

Objective To evaluate the effects of leukocyte-depleted allogeneic blood transfusion on perioperative cellular immunity in children.Methods Three hundred and fifty-nine ASA Ⅰ or Ⅱ children (aged 3 months-14 years and weighing 5-74 kg) requiring allogeneic blood transfusion during operation were randomly divided into two groups:163 children receiving normal allogeneic blood transfusion (control group,group C) and 196 children receiving leukocyte-depleted allogeneic blood transfusion (group D).Blood samples were collected from the peripheral vein before blood transfusion,and 2 and 6 days after blood transfusion for determination of the levels of CD3+,CD4+,CD8 +,and CD56+ by flow cytometry.CD4+ /CD8+ ratio was calculated.The volume of allogeneic blood transfusion during operation,the duration of operation,postoperative drainage,antibiotic administration,hospital stay and the incidence of postoperative infection were recorded.Rssults The levels of CD3+,CD4+,CD56+ and CD4+/CD8+ ratio significantly increased at 6 days after blood transfusion while the duration of postoperative drainage,postoperative antibiotic administration,hospital stay and the incidence of postoperative infection significantly decreased in group D compared with group C.Conclusion Leukocyte-depleted allogeneic blood transfusion is helpful in improving the postoperative cellular immunity in children.