Objective To study the role of bone morphogenetic protein-7 in the osteogenic differentiation of periosteal cellsin vitro. Methods Periosteal cells, obtained from adult tibial periosteum, were cultured by routine method in vitro, and divided into two groups. One group cultured with BMP7 and the supplements of 100 nmol dexametasone, 10 mmol b-glycerophosphate and 50 mg/mL L-ascorbic acid (BMP7 group), the other cultured with the supplements alone as the control (control group). Ultrastructure and morphological changes of periosteal cells were observed by contrast phase microscope and electron microscope. In order to test the expression of markers of osteoblastic differantiation in periosteal cells, involved mineralized node and alkaline phosphatase. Each group was tested at the time of 5 d, 10 d, 15 d, 20 d, respectively, using ALP kit stain and Von Kossa stain with 3 samples at each time. Results The periosteal cells cultured by routine method and induced into osteoblast differentiation with BMP7 were both growing well, in vitro. Microscope observations showed that the periosteal cells were spindle-shaped, well-stacked, transparent and three-dimensional in the early stage, and cube-shaped or puncheon shaped in the mitotic phase, gradually became wide shuttle and irregular shape with a lot secretion in telophase. The positive cells were visible by the ALP kit staining and Von Kossa staining of calcium nodules at 5 d, 10 d, 15 d and 20 d in both groups.A difference of positive rate at each time point was found between BMP7 group and control group at 5 d, 10 d, 15 d, 20 d, and the difference was statistically significant (P ＜ 0.01). Conclusion It displayed well regeneration and osteogenesis ability in the periosteal cell. BMP7 has definite osteo-inductive activity, which can obviously enhance the proliferation and ossifyng differentiation of periosteal cells.

Objective To analyze the contrast agent back flow,balloon ejection and pain which appeared during the examination process,and define the advantages and disadvantages of different rates of high pressure injector to take hysterosalpingography (HSG) examination.Methods 229 patients who took HSG examination in our department were divided into two groups randomly.The rate of conventional group was 0.3 mL/s,and low rate group was 0.15 mL/s.The contrast agent back flow,balloon ejection and pain which appeared on the examined patients of two groups were recorded.T test was adopted to compare the age difference of two groups.χ2 test was used to compare the differences of diagnosis results,contrast agent back flow and balloon ej ection of two groups.Wilcoxon rank sum test was used to compare the pain difference of two groups.Results Two groups had no significant differences on age, diagnosis results and contract agent back flow (t=1.237,χ2=0.004,χ2=1.521,P>0.05).Balloon ejection of two groups had significant difference(χ2=17.319,P<0.05).The appeared pain of two groups had significant difference (U=-2.337,P<0.05).Conclusion Using high pressure injector to inject contrast agent with low flow rate is better for HSG.

Objective:To explore the efficacy and safety of FOLFOX4 regimen hepatic arterial infusion chemotherapy (HAIC) combined with 125I seed implantation or gamma knife in patients with primary liver cancer and type Ⅲ portal vein tumor thrombosis (PVTT). Methods:The clinical data of 116 patients with primary liver cancer and type Ⅲ PVTT from January 2016 to June 2018 in the Sixth Medical Center of PLA General Hospital were retrospectively analyzed. Among them, 56 patients were treated with FOLFOX4 regimen HAIC combined with 125I seed implantation in PVTT (observation group), and 56 patients were treated with FOLFOX4 regimen HAIC combined with gamma knife in PVTT (control group). The clinical efficacy, adverse reactions, liver function, tumor serum markers, quality of life and portal venous pressure were compared between 2 groups. The patients were followed up to June 2021, the progression-free survival (PFS) time, overall survival (OS) time and 1-, 3-year survival rate were recorded. Results:The objective response rate and disease control rate in observation group were significantly higher than those in control group: 21.43% (12/56) vs. 7.14% (4/56) and 50.00% (28/56) vs. 30.36% (17/56), and there were statistical differences ( P<0.05). The aspartate transaminase (AST), alanine transaminase (ALT), total bilirubin, alpha fetoprotein (AFP), carcinoembryonic antigen (CEA) and tumor supplied group of factor (TSG) after treatment in observation group were significantly lower than those in control group: (58.24 ± 7.16) U/L vs. (67.81 ± 8.39) U/L, (56.39 ± 5.42) U/L vs. (62.87 ± 6.83) U/L, (21.21 ± 4.32) μmol/L vs. (25.88 ± 4.71) μmol/L, (32.98 ± 6.15) μg/L vs. (45.06 ± 7.24) μg/L, (2.39 ± 0.22) μg/L vs. (3.17 ± 0.26) μg/L and (57.81 ± 5.67) kU/L vs. (66.19 ± 5.45) kU/L, and there were statistical differences ( P<0.01). The quality of life questionnaire core-30 (QLQ-C30) score after treatment in observation group was significantly higher than that in control group: (68.13 ± 6.95) scores vs. (64.49 ± 6.73) scores, the portal venous pressure after treatment in observation group was significantly lower than that in control group: (31.85 ± 3.89) cmH 2O (1 cmH 2O = 0.098 kPa) vs. (35.37 ± 4.23) cmH 2O, and there were statistical differences ( P<0.01). There was no statistical difference in incidence of adverse reactions between 2 groups ( P>0.05). Two cases were lost in observation group and 3 cases in the control group; the PFS time and OS time in observation group were significantly longer than those in control group: (13.36 ± 2.85) months vs. (11.76 ± 2.60) months and (23.36 ± 4.37) months vs. (20.72 ± 3.96) months, and there were statistical differences ( P<0.01); the 1-year survival rate in observation group was significantly higher than that in control group: 70.37% (38/54) vs. 50.94% (27/53), and there was statistical difference ( P<0.05); there was no statistical difference in 3-year survival rate between 2 groups ( P>0.05). Conclusions:The anti-tumor effect and the improvement effect on liver function in patients with primary liver cancer and type Ⅲ PVTT treated with FOLFOX4 regimen HAIC combined with 125I seed implantation are better than combined gamma knife, and it has high safety and longer survival time.

Objective:To investigate the relationship between double mutations of myosin heavy chain gene (MYH6) p.Gly743Arg and p.Glu1389Lys and the cardiac phenotype.Methods:Patients carrying double mutations in the MYH6 gene p.Gly743Arg and p.Glu1389Lys were screened from 52 unrelated left ventricular hypertrophy (LVH) who were admitted to the Second Hospital of Chongqing Medical University from 2015 to 2020, and the genetic testing of peripheral blood of patients by second-generation whole-exome sequencing assay technology and genomic DNA of their family members Sanger sequencing was performed to validate the genomic DNA of the family members. The cardiac phenotype was evaluated by electrocardiogram, coronary computed tomography angiography (CTA), echocardiography, and cardiac magnetic resonance imaging (MRI) as adjuncts.Results:All whole-exome gene were detected in 52 unrelated patients with LVH, of which 1 patient (1.9%) had double mutations in MYH6 gene p.Gly743Arg and p.Glu1389Lys (proband). Two members of the maternal line of this patient carried p.Glu1389Lys mutation, but there was no obvious clinical phenotype. Two members of the paternal line carried p.Gly743Arg mutation and had obvious clinical phenotype of bradycardia, but there was no LVH. The male proband, aged 21 years old, presented with LVH and sinus bradycardia but no coronary artery stenosis on CTA before treatment, MRI showed that the left ventricular end diastolic diameter was 58 mm. After treatment with angiotensin receptor-enkephalinase inhibitor (ARNI), electrocardiogram showed that the heart rate increased significantly (from 43 bpm to 72 bpm). Echocardiography showed that the left ventricular end diastolic diameter decreased significantly (from 60 mm to 49 mm).Conclusions:The p.Glu1389Lys mutation of the MYH6 gene may not manifest the phenotype of heart disease. MYH6 gene p.Gly743Arg mutation may be manifested asymptomatic sinus bradycardia, but there is no LVH phenotype. The cardiac disease phenotype caused by the double mutations of p.Gly743Arg and p.Glu1389Lys in the MYH6 gene is more obvious. Asymptomatic LVH and sinus bradycardia can appear in adolescence, but the LVH phenotype can be reversed in a short period of time after ARNI treatment.

Objective To analyze the CT signs of acute blunt or penetrating intestinal injury,and to improve the diagnostic accuracy of multi spiral CT for intestinal injury.Methods The CT and clinical data of 63 patients with intestinal injury confirmed by clinical surgical exploration who underwent emergency CT scan were collected,and the CT findings and surgical findings were comparatively analyzed.Results There were 63 cases of intestinal injury,of which 26 cases were complicated with mesenteric injury.The direct CT signs of intestinal injury included intestinal wall thickening sign and intestinal discontinuity sign,which accounted for 64%(40/63)and 17%(10/63),respectively.The indirect CT signs of intestinal injury included intraperitoneal/retroperitoneal gas sign,intraperitoneal/retroperitoneal effusion sign,intramural air,and portal venous gas,which accounted for 72%(45/63),88%(55/63),7%(5/63)and 5%(3/63),respectively.Conclusion Recognizing the CT signs of intestinal injury,such as intestinal wall thickening sign,intestinal discontinuity sign,intraperitoneal/retroperitoneal gas sign,intraperitoneal/retroperitoneal effusion sign,intramural air,and portal venous gas can help to make the early and correct diagnosis of intestinal injury if combined with clinical practice.

Objective:To develope an automatic volumetric modulated arc therapy (VMAT) planning for rectal cancer based on a dose-prediction model for organs at risk(OARs) and an iterative optimization algorithm for objective parameter optimization.Methods:Totally 165 VMAT plans of rectal cancer patients treated in Peking University Cancer Hospital & Institute from June 2018 to January 2021 were selected to establish automatic VMAT planning. Among them, 145 cases were used for training the deep-learning model and 20 for evaluating the feasibility of the model by comparing the automatic planning with manual plans. The deep learning model was used to predict the essential dose-volume histogram (DVH) index as initial objective parameters(IOPs) and the iterative optimization algorithm can automatically modify the objective parameters according to the result of protocol-based automatic iterative optimization(PBAIO). With the predicted IOPs, the automatic planning model based on the iterative optimization algorithm was achieved using a program mable interface.Results:The IOPs of OARs of 20 cases were effectively predicted using the deep learning model, with no significantly statistical difference in the conformity index(CI) for planning target volume(PTV)and planning gross tumor volume(PGTV)between automatic and manual plans( P>0.05). The homogeneity index (HI) of PGTV in automatic and manual plans was 0.06 and 0.05, respectively( t=-6.92, P< 0.05). Compared with manual plans, the automatic plans significantly decreased the V30 for urinary bladder by 2.7% and decreased the V20 for femoral head sand auxiliary structure(avoidance)by 8.37% and 15.95%, respectively ( t=5.65, 11.24, P< 0.05). Meanwhile, the average doses to bladder, femoral heads, and avoidance decreased by 1.91, 4.01, and 3.88 Gy, respectively( t=9.29, 2.80, 10.23, P< 0.05) using the automatic plans. The time of automatic VMAT planning was (71.49±25.48)min in 20 cases. Conclusions:The proposed automatic planning based on dose prediction and an iterative optimization algorithm is feasible and has great potential for sparing OARs and improving the utilization rate of clinical resources.

BACKGROUND:The treatment for bacterial biofilms after internal fixation surgery is a very difficult problem in clinic.It is a great significance to establish an animal model of irrigation for treating bacterial biofilms in the early stage after internal fixation surgery. OBJECTIVE:To establish an animal model for treating bacterial biofilms with different drugs through irrigation in early stage after internal fixation surgery. METHODS:Six New Zealand white rabbits were selected.Bilateral femoral surfaces were exposed and drilled holes were made,and bone plates colonized with Pseudomonas aeruginosa(experimental group)and blank bone plates(blank control group)were implanted around the drilled holes on one side,and two drainage tubes were retained and fixed to serve as the"inlet"and"outlet,"respectively.The model was immersed for a certain period of time after simulated perfusion before rinsing.After the simulated irrigation,the plates were soaked for a certain time before washing.At 5 days postoperatively,the rabbits were observed for body temperature,wound condition,bacterial culture of drainage fluid,and crystalline violet staining and scanning electron microscopy of the bone plate. RESULTS AND CONCLUSION:Six rabbits had difficulty in moving the affected limbs after surgery and showed elevated body temperature at 2-4 days after surgery.Local swelling could be touched at some wounds in the experimental group,and the wounds in the blank control group healed well.The results of bacterial culture of drainage fluid showed that Pseudomonas aeruginosa diffused or spread in the experimental group.At 5 days after surgery,the plate in the experimental group became purple shown by crystalline violet staining,and the absorbance value at 570 nm detected by the microplate reader was 2.621±0.088,indicating the presence of bacteria.Scanning electron microscopy at 5 days after surgery showed that a large number of bacterial microcolonies appeared on the surface of the plate in the experimental group,forming a highly inhomogeneous three-dimensional structure similar to the"mushroom-like"and"tower-like"structures,with filamentous water channels connecting the"mushroom-like"structures,which were typical biofilm structures with high densities,while no obvious colonies were seen in the blank control group.Overall,this animal model simulates the state of infected biofilm formation due to early infection after internal fixation and provides an available method of irrigation with different drugs.

【Objective】 To establish a method for detecting human immunoglobulin Fc function based on surface plasmon resonance technology. 【Methods】 Based on the characteristic that FcγRI can be binded to the Fc segment of IgG, the affinity constant of the sample was detected by surface plasmon resonance, and its Fc function was the KD ratio of the sample to the standard. The method was validated for specificity/specificity, precision and robustness. The method and the pharmacopoeia method were used to detect the Fc function of 30 human immunoglobulins, and the correlation and consistency of the detection results were analyzed. 【Results】 The method validation results showed that this method has strong specificity/specificity (t values were 0.15, 0.22, both P>0.05), good precision (CV value 5.37%~10.69%) and good robustness (CV value 10.06%). The detection results of this method and the pharmacopoeia method have high correlation (r=0.96, P<0.05) and high consistency (Bias-2.060, 95% Limits of Agreement-5.628~1.508). 【Conclusion】 A method for detecting human immunoglobulin Fc function based on surface plasmon resonance has been successfully established.

Adventitia ; Atherosclerosis/genetics* ; Cell Communication ; Humans ; Hyperhomocysteinemia/genetics* ; Sequence Analysis, RNA