Objective To compare transfection properties of two different fluorescently labeled PIK3CA siR-NA and to screen out PIK3CA siRNA with high transfection efficiency and strong anti-quenching ability.Methods Two different fluorescently labeled PIK3CA siRNA was transfected into gastric cancer cell BGC-823 by Lipofectamine 2000.The distribution and quenching of fluorescence were observed by inverted fluorescence microscope.Image J software was used to analyze their transfection efficiency.Real-time quantitative PCR was performed to detect the effect of different fluorescently labeled PIK3CA siRNA on PIK3CA mRNA expression.Results The transfection efficiency and anti-quenching ability of two different fluorescently labeled PIK3CA siRNA were different under the same transfection conditions.The transfection efficiency showed no significant differences between Cy3 or FAM labeled PIK3CA siRNA and negative control siRNA(P ＞ 0.05),but the transfection efficiency of Cy3 labeled PIK3CA siRNA and negative control siRNA was significantly higher than the FAM labeled (P ＜ 0.05).Inhibitory efficacy of target mRNA expression induced by Cy3 labeled PIK3CA siRNA was significantly higher than that of FAM labeled PIK3CA siRNA.Conclusion Cy3 labeled PIK3CA siRNA could act as a good tracer and provide an important evidence for further construction of Cy3 labeled PIK3CA siRNA nanoparticle.

Objective To explore the CT and MRI performances of primary extragonadal seminoma .Methods The imaging find‐ings of primary extragonadal seminoma in 16 patients proved by histopathology were analyzed retrospectively .The tumor location , size ,contour ,periphery ,density or signal and contrast enhancement patterns were evaluated ,and these were compared with the pathological results .Results The lesions in 16 patients were solitary including the ovoid lesions in 6 and lobulated ones in 10 .They were located in intracranial area in 10 patients ,anterior middle mediastinum in 4 and abdomen in 2 .Mean size of the tumors was(7 .0 ± 5 .6) cm (ranging from 1 .3 cm to 18 .6 cm) .Unenhanced CT in 13 patients showed the soft tissue masses with necrosis in 8 ,stip‐pled calcification in 2 and hemorrhage in 2 .Enhanced CT in 6 patients showed homogeneity in 2 and heterogeneity in 4 ,and the de‐grees of enhancement increased gradually .The plain and enhanced MRI in 10 patients showed iso‐or slight hypointensity on T1WI and slight hyperintensity on T2 WI in 8 lesions ,heterogeneous enhancement in 7 and slight enhancement in one ,peripheral enhance‐ment in 2 due to central hemorrhage .Pathology showed most of the lesions were well‐vascularized with inner cystic change .Tumor stroma was divided by the fibrovascular tissue .Conclusion The imaging findings of primary extragonadal seminoma are not specific , CT ,MRI can demonstrate the invasive extent accurately .

Objective To explore the feasibility and effect of laparoscopic Ladd's surgery for the treatment of intestinal malrotation in infants.Methods The clinical data of 50 infants with intestinal malrotation who were admitted to the Wuhan Medical & Health Center for Women and Children from January 2011 to December 2013 were retrospectively analyzed.Of 50 infants, 27 infants receiving the open Ladd's surgery were allocated into the open surgery group and 23 infants receiving the laparoscopic Ladd's surgery were allocated into the laparoscopy group.The operation time, time to anal exsufflation, duration of hospital stay, postoperative incision infection and intestinal obstruction in the 2 groups were observed.All the patients were followed up by outpatient examination or telephone interview till March 2014.The measurement data with normal distribution were presented as x-± s and analyzed by the t test, and count data were analyzed using the chi-square test or Fisher exact probability.Results The operation time, time to anal exsufflation and duration of hospital stay were (69 ± 7)minutes, (41 ±9)hours and (10.4 ± 2.4)days in the open surgery group, which was significantly different from (92 ± 13)minutes, (28 ±5)hours and (6.4 ± 1.5) days in the laparoscopy group (t =6.21, 16.50, 6.34,P ＜ 0.05).Two infants had incision infection and 1 infant had intestinal obstruction in the open surgery group.There was no patient with complications in the laparoscopy group.All the patients were followed up for the median time of 13 months (range, 6-24 months), with a good survival and no other symptoms.Conclusion Laparoscopic Ladd's surgery is safe and feasible compared with open surgery, and it could be used as a prior operation method for treatment of intestinal malrotation in infants.

OBJECTIVETo investigate the effect of PIK3CA/siRNA chitosan nanoparticle on the invasiveness of gastric carcinoma and the potential value of PIK3CA/siRNA chitosan nanoparticle in suppressing the metastasis of gastric carcinoma.

METHODSGastric cancer cells were treated with PIK3CA/siRNA nanoparticle (with a diameter of 350 nm), and the efficiency of PIK3CA gene interference was evaluated using Western blotting and real-time PCR. The changes of the invasive capacity of the treated cells was assessed with Transwell assay.

RESULTSPIK3CA/siRNA chitosan nanoparticle efficiently lowered the expression level of PIK3CA and significantly decreased the invasion of BGC823 cells.

CONCLUSIONPIK3CA gene interference mediated by PIK3CA/siRNA chitosan nanoparticle can decrease the invasive capacity of gastric cancer cells in vitro.

Cell Line, Tumor ; Cell Proliferation ; Chitosan ; Class I Phosphatidylinositol 3-Kinases ; Humans ; Nanoparticles ; Phosphatidylinositol 3-Kinases ; genetics ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction ; Stomach Neoplasms ; pathology

Objective: To determine the cut-off values of Ki-67 labeling index (LI) in the histological grading of follicular lymphoma (FL). Methods: Clinicopathological data of 350 FL patients diagnosed at Beijing Friendship Hospital from June 2014 to January 2016 were analyzed retrospectively by quantitative evaluation and statistical analysis of Ki-67 LI. Results: Of the 350 patients with FL, the male and female ratio was 1.1 and the average age was (50.2±14.0) years with a median age of 51 years (range 4 to 82 years). The tumors were graded as grade Ⅰ-Ⅱ in 215 cases (61.4%), grade Ⅲ A in 105 cases (30.0%), and grade Ⅲ B in 30 cases (8.6%). The average Ki-67 values were (22.8%±8.3%) for the FL low (grade Ⅰ-Ⅱ) and (50.4%±10.7%) for high grade (Ⅲ A and Ⅲ B) and were statistically significant by Mann Whitney U test (P<0.01). Receiver operated characteristic curve analysis showed that the best diagnostic cut-off value of low grade FL was 35% (sensitivity of 96.3% and specificity of 93.3%) with the largest area under curve (AUC=0.990, P<0.01, 95%CI for 0.982-0.998). According to the analysis of four lattice diagnostic tests, Ki-67 LI >40% was an important factor (χ²=230.733, P<0.01) in predicting high grade FL. When the cut-off value of Ki-67 LI was set at 40%, high grade LF could be diagnosed with the greatest sensitivity (98.1%) and specificity (87.7%). Moreover, a significant correlation between the Ki-67 LI and the pathological grade of FL (r=0.836, P<0.01) was observed. Conclusions Ki-67 LI of below a cut-off value of 35% is a reliable indicator of low grade FL.Ki-67 over 40% is consistent with high grade FL. These Ki-67 cut-off values may serveas an important auxiliary indicator in the grading of FL.

OBJECTIVETo investigate changes in renal function, urine N-acetyl-beta-D-glucosaminidase enzyme (N-AG),liver function, myocardial enzymes, the pathology of renal damage and the mechanism of acute renal failure (ARF) associated with fish gall bladder poisoning.

METHODSEleven patients with acute fish gall bladder poisoning were consecutively admitted to our hospital from September 1997 to October 1999. Renal function, urine N-AG enzyme, liver function, and myocardial enzymes were assayed before and after treatment. One patient consented to a kidney biopsy and the pathology of renal damage was observed under light and electron microscopes.

RESULTSAll patients had multiple organ dysfunction syndrome (MODS) and 11 patients suffered from ARF. Ten patients had liver dysfunction, ten patients had poisonous myocarditis, and 8 patients had gastrointestinal dysfunction. Renal function, urine N-AG enzyme, liver function, and myocardial enzymes were significantly improved after treatment compared with those of before treatment (P < 0.05). Kidney biopsy showed that the main damage site was the proximal renal tubule. All eleven patients recovered and were discharged from the hospital.

CONCLUSIONSIngestion of fish gall bladder leads to kidney damage, as well as liver, heart and gastrointestinal tract injury. The mechanism of acute renal function failure is the serious tubular damage, confirming the location of kidney damage. Necrosis of the proximal tubules plays an important role in the development of ARF. Immediate hemodialysis is the most effective treatment.

Adult ; Aged ; Animals ; Female ; Fishes ; Foodborne Diseases ; etiology ; Gallbladder ; Humans ; Male ; Middle Aged ; Multiple Organ Failure ; etiology

Objective:To investigate the clinicpathological features of basal cell type dysplasia of the esophagus.Methods:The clinicopathological data of 71 cases of basal cell type dysplasia of esophagus were collected at the People′s Liberation Army Joint Logistics Support Force 989 Hospital, from 2009 to 2019, and the histomorphologic characteristics and immunophenotype were evaluated. The relevant literature was reviewed.Results:The ratio of male to female patients was 1.6∶1.0, and the median age was 65 years (range 48-81 years). The tumors were located in the upper segment of the esophagus in four cases (5.6%), the middle segment in 54 cases (76.1%), and the lower segment in 13 cases (18.3%).The median maximal tumor diameter was 12.0 mm (range 3-42 mm). According to Paris Classification, 0-Ⅱb accounted for 42.3% (30/71) of the cases. Under endoscope, the lesions were reddish with abnormal mucosal microvessels. Histologically, the neoplastic cells were small, with a high nuclear-cytoplasmic ratio, similar to basal cells, and uniform in morphology. The structural atypia was characterized by dense and disordered tumor cells, loss of basal cell polarity, and absence of normal squamous differentiation gradient. In 10 cases, the tumors were confined to the lower part of the epithelium. The tumor cells were smaller and more uniform in shape, and extend to the superficial lamina propria. Sixty-one tumors involved at least the entire layer of the upper cortex. There were 31 cases of neoplasms with superficial invasive carcinoma. The types of neoplasms included typical squamous cell carcinoma, basaloid squamous cell carcinoma, small cell neuroendocrine carcinoma, squamous cell carcinoma with sebaceous adenoid carcinoma, and differentiation of glandular/ductal epithelioid carcinoma. Immunohistochemical staining showed that the mutant expression rate of p53 protein was 41.5% (17/41). All 41 cases (100.0%) showed abnormal distribution pattern of Ki-67. According to the initial pathologic diagnosis, there were 18 cases of low grade dysplasia, 12 cases of atypical epithelial cells, and 41 cases of high grade dysplasia and superficially invasive carcinoma.Conclusions:Basal cell type dysplasia has unique morphologic characteristics and represents a tumor subtype in the morphologic lineage of esophageal squamous dysplasia. Tumor cells of basal cell type dysplasia, especially those distributed only in the lower part of the stratified squamous epithelium, may be tumor stem cells at the earliest stage of esophageal carcinogenesis and have multidirectional differentiation potential. When the tumor is confined to the lower part of the stratified squamous epithelium, it does not meet the diagnostic criteria for esophageal squamous dysplasia as defined by the current WHO classification.

Objective: To study the clinical reference pulpal blood flow(PBF) values detected by laser doppler flowmetry(LDF) in healthy young population and to analyze their possible affected factors. Methods: Undergraduate students at the age of 17-23 years were enrolled. PBF of 12-22 were detected by LDF based on the standard procedure. Difference of the test results between different sex was analyzed by T test and variance homogeneity test, and the correlation with age was analyzed by the chi-square test, and the difference between the different teeth was analyzed by the random group analysis. Results: 400 students(250 males and 150 females with the average age of 19. 83 years) met the inclusion criteria. The clinical reference values of PBF of different anterior teeth were obtained by the detection of LDF. For the same tooth, PBF values of females were higher than that of males (P＜ 0. 05). PBF values of different ages shared no statistical significance(P＞ 0. 05). For the same gender, PBF values of middle incisor were higher than that of lateral incisors(P＜ 0. 05). Conclusion: The determination of the clinical reference values of PBF detected by LDF may promote the clinical use of this technology.

BACKGROUND:There are many treatment methods for knee osteoarthritis,among which electroacupuncture,as an important non-drug treatment,is effective in the treatment of knee osteoarthritis,but its exact mechanism is not clear. OBJECTIVE:Effect of electroacupuncture on the expression of p53 and P21 in articular cartilage and subchondral bone of aged rats with knee osteoarthritis. METHODS:Eight 6-month-old male Sprague-Dawley rats were included in the young group and sixteen 24-month-old male Sprague-Dawley rats were randomly divided into old group(n=8)and electroacupuncture group(n=8).The rats in the electroacupuncture group received electroacupuncture stimulation once a day,5 days a week,for 8 continuous weeks,and the other two groups did not do any treatment.Eight weeks later,the level of type Ⅱ collagen C-terminal peptide in peripheral blood was detected by ELISA,the morphology of left knee cartilage and subchondral bone was observed by safranin O-fast green staining,the degree of knee cartilage degeneration was evaluated by modified Mankin's score,the microstructure of left knee cartilage and subchondral bone was detected by micro-CT,and the expression levels of matrix metalloproteinase 13,P53,P21 Mrna and protein were detected by RT-PCR and western blot respectively. RESULTS AND CONCLUSION:Compared with the young group,the level of C-terminal peptide of type Ⅱ collagen in the peripheral blood was increased in the old group(P<0.05).The micro-CT results showed that the bone volume fraction,bone mineral density and the number of bone trabeculae were decreased in the old group compared with the young group(P<0.05),while the trabecular separation increased(P<0.05).Safranin O-fast green staining showed that in the old group,the surface layer of cartilage was uneven with fissures,the morphology of chondrocytes was irregular and stained unevenly,the boundary between the cartilage and subchondral bone was blurred,and the matrix loss was serious.The Mankin's score was higher in the old group than the young group(P<0.05).The expression of matrix metalloproteinase 13,P53,P21 at Mrna and protein levels increased in the old group compared with the young group(P<0.05).Compared with the old group,electroacupuncture decreased the level of C-terminal peptide of type Ⅱ collagen(P<0.05),increased the bone volume fraction,bone mineral density and the number of bone trabeculae(P<0.05),and decreased the trabecular separation(P<0.05).Safranin O-fast green staining showed that in the electroacupuncture group,the surface of cartilage was smooth and red staining was uniform,and the cell morphology and structure were between the young group and the old group.Following electroacupuncture treatment,the Mankin's score(P<0.05),matrix metalloproteinase 13 and P21 Mrna expression(P<0.05),and matrix metalloproteinase 13 and P53 protein expression decreased(P<0.05),while there was a decreasing trend of P53 Mrna and P21 protein expression,but with no statistical significance(P>0.05).To conclude,electroacupuncture may delay articular cartilage degeneration and subchondral osteoporosis in aged rats by inhibiting the expression of P53 and P21,so as to protect joints and delay joint aging.