Objective To investigate the effects on self-renewal of pancreatic cancer stem cells by inhibiting hedgehog signaling pathway through cyclopamine. Methods PANC1 stem cells, PANC1 adherent cells and immortalized pancreatic ductal epithelial H6C7 cells were treated with 0.5, 1, 2, 5, 10 mol/L of cyclopamine for 24, 48, 72 h. The expression of Smo mRNA and Gli1 mRNA were detected by real-time PCR.Cell growth viability was measured by CCK 8. Cell cycle and apoptosis were determined by flow cytometry.Results Seventy-two hours after cyclopamine treatment, the Smo mRNA expressions of PANC1 stem cells,PANC1 adherent cells and H6C7 cells were 1,0.83 and 2.61; the expressions of Gli mRNA were 57.27,26.35,1; the inhibitory rates were ( 37.85 ± 13.69 ) %, ( 8.53 ± 4.43 ) %, (43.55 ± 28.98 ) %. Compared with PANC1, the expressions of Smo mRNA, Gli1 mRNA and the inhibitory rate of PANC1 stem cells significantly increased ( P ＜ 0.05 ). The proportion of G1 stage of PANC1 stem cells significantly decreased from (67.41 ±6.35)% to (36.53 ±6.03)% (P ＜0.05), and the apoptosis decreased from (10.95 ±5.68) % to ( 5.73 ± 1.42 ) % ( P ＞ 0.05 ). The proportion of G1 stage of PANC1 cells significantly decreased from ( 67.64 ± 6.88 ) % to ( 53.13 ± 1.10 ) % ( P ＜ 0.05 ); the apoptosis decreased from ( 12.08 ±4.12)% to (5.66 ± 1.33)% (P ＞0.05). While both the proportion of G1 stage and apoptosis of H6C7 cells was not significantly different. Conclusions Cyclopamine can inhibit the proliferation of PANC1 stem cells via blocking hedgehog signal pathway, and the mechanism may not be associated with cell apoptosis.

Objective To establish the method of suspension culture for stem cells from human pancreatic cancer cell line PANC1.Methods PANC1 cells were cultured in serum-free medium under floating-culture system.Tumor cell spheres were observed by optical microscope.Expression of CD133 and cell cycle were detected by flow cytometry.Cancer stem cells were induced to differentiate with 10%FBS,and expression of CK18,was evaluated with immunofluorescence microscope.Spheres cells were injected into the subcutaneous space of NOD/SCID rat and tumor formation was monitored weekly.Results PANC1 cells could form the stem cells spheres,and the rate of sphere formation was stable between 4%0 and 5%0 after 20 passages in vitro.The expression of CD133(5.91±0.7%)and proportion of G0/G1 phase cell(80.99±2.60%)was significantly increased in spheres cells compared with parental PANC1 cells(1.44±0.52%and 69.01±5.03%),and the difference was statistically significant(P＜0.05).When these spheres cells were cultured in media with serum,these cells gradually returned to the status of parental cells and expressed CK18,2×103 sphere cells injection could initiate tumor fornmtion in NOD/SCID rat.Conclusions Tumor spheres stem cellscould be generated under serum-free floating-culture system.The sphere cells possessed the capacities of self renew,difierentiation,and tumorigenic potential.

Objective To study the effect of story inquiry in the care to patients undergoing laparoscopic cholecystectomy. Methods Toally 39 patients to undergo laparoscopic cholecystectomy during January to April 2017 were set as control group and another 39 patients to to undergo laparoscopic cholecystectomy during May to August 2017 were set as treatment group. The former received routine nursing care and the latter was treated with the story inquiry including five stages of preparation,story sharing,dialogue inquiry,situational association and comfort making,apart from routine nursing care.The two groups were compared in terms of anxiety, depression and sleep quality. Result The scores of depression and anxiety and sleep quality after laparoscopic cholecystectomy in the experimental group were significantly lower than those in the control group (P＜0.001). Conclusion The effect of emotional comfort and sleep quality improvement can be improved by using the method of story inquiry for the patients undergoing laparoscopic cholecystectomy.

Objective: To analyze the clinical response in cases of multi-victims bitten by a dog in Beijing (one of whom was HIV positive), and to explore the route and prognosis of dual exposure to rabies virus and HIV. Methods: Descriptive epidemiological methods were used to analyze the exposed cases, the post-exposure prophylaxis(PEP) of rabies and HIV and the follow-up outcomes. Results: After six months-follow up, there was no rabies case was found among those nine victims bitten by the suspected dog. In addition, six HIV-negative victims who had been exposed to HIV via dog saliva were consistently tested to be HIV negative in the follow up period. Conclusions In cases with multi-victims bitten by one dog, except exposure to rabies virus, it is necessary to be alert to the presence of HIV, HBV or HCV infection in sequential victims. Therefore, the risk of the potential spreading of other blood-borne infectious pathogens should be assessed. It is important to initiate PEP as early as possible.

Objective: To understand the prophylaxis of population exposed to rabies, and provide a basis for prevention and control of rabies. Methods: The registration data of 1 019 cases with rabies exposure in Beijing Ditan Hospital Affiliated to Capital Medical University from Mar, 2017 to Feb, 2018 were analyzed using the descriptive epidemiologic method. Results: Among the 1 019 cases of rabies exposure, the sex ratio of men to women was 0.90∶1, with the highest proportion of them were between 26 and 46 years of age old, accounting for 46.5%. Rabies exposure reached its peak (46.9%) between June and September. In addition, 63.3% of the injuries were caused by dogs and 36.7% by cats. Most of the wounds (55.1%) occurred in hands; and the second was lower limbs (26.6%). Most of the cases (60.7%) had grade II wounds; followed by grade III wounds (39.1%); 69.3% of the cases completed immunization with rabies vaccine. In grade III exposure, 73.7% of the patients were immunized with rabies immunoglobulin. Most of the persons of grade III exposure (73.7%) received inoculation with human rabies immunoglobulin (HRIG). Conclusions Measures to control rabies should be focused on the management of dogs and cats and the standardization of rabies post-exposure prophylaxis.

Objective:To evaluate the feasibility of Full-stack Smart Pelvic Floor Ultrasound (FSPFU) software in the acquisition and measurement of the minimal levator hiatus (LH).Methods:Transperineal pelvic floor ultrasonography was performed in 119 women of 6-month postpartum from Nov.2020 to Jan.2021 of Shenzhen Second People′s Hospital. Mid-sagittal plane of pelvic floor was set as the initial plane, and the three-dimensional volume data was acquired. The dataset was stored in the machine. The offline volume data was manually adjusted to obtain the minimal LH images and measured by four physicians (two junior physicians as the D1 group and two senior physicians as the D2 group). For comparison, the results were also obtained using the fully automated method—the FSPFU software by a junior physician (the D3 group). The obtained parameters of minimal LH included area, circumference, anterioposterior diameter, transverse diameter, left and right levator-urethral gap distance. Analysis time was recorded for each group. The contours of minimal LH were outlined by three groups and the overlapping rate was calculated. The quality of the resulted images was evaluated and scored by another two senior physicians(A and B) independently.Results:The D3 group had a significant shorter analysis time compared with the other two groups, and the D1 group took a longer time than the D2 group, regardless of the cystocele severity (D1: 82.97 s, D2: 62.51 s, D3: 2.71 s, all P<0.05). The intergroup agreements and correlations of the minimum LH area were good (all ICC>0.85, rs>0.70, P<0.001) and the outlined contours were largely overlapped (>92%). There was no significant difference in image quality among the three groups(all P>0.05). Conclusions:FSPFU software can automatically obtain and measure the minimum LH in an efficient and accurate way, which can improve the effectiveness of the present pelvic floor examination. FSPFU software can be an useful tool in the diagnosis of pelvic floor dysfunctional diseases.

【Objective】 To evaluate the safety and effectiveness of Perioperative Transfusion Trigger Score (POTTS) in guiding surgical patients blood transfusion intraoperatively and postoperatively. 【Methods】 A total of 900 patients(perioperative Hb 60~100 g/L) from December 2017 to March 2021 were collected, including 251 males and 649 females, with ASA grading Ⅰ~Ⅳ, and randomly divided into experimental group and controls. In the experimental group, the allogeneic RBC transfusion trigger(Hb threshold) and transfusion units in anemia patients was determined by POTTS. While those in the controls were decided by physicians according to current guidelines concerning transfusion. The proportion and units of allogeneic RBC transfusion, the incidence of postoperative complications, the mortality of hospitalization and discharge for 4 weeks, Hb value, healing of surgical incision, ICU admission rate and ICU length of stay, length of hospitalization, etc were recorded. 【Results】 The proportion of allogeneic RBC transfusion in the experimental group(35.3%)were less than the controls(42.2%)(P<0.05). The units of allogeneic RBC transfused, incidence of postoperative complications and mortality in hospital and 4 weeks after surgery, healing of surgical incision (grade A/ grade B/ grade C), the Hb level 24 h after surgery and at discharge, the ICU admission rate and ICU length of stay, length of hospitalization were not significantly different between the groups. 【Conclusion】 The peri-operative allogeneic red blood cells transfusion guided by POTTS can reduce the proportion of allogeneic RBC transfusion, and is safe and effective.