Effects of the kynurenine pathway on the osteogenic differentiation of periodontal ligament stem cells / 中华口腔医学杂志

Objective: To explore the effect of kynurenine pathway on the osteogenic differentiation of periodontal ligament stem cells (PDLSC). Methods: Unstimulated saliva samples were collected from 19 patients with periodontitis (periodontitis group) and 19 periodontally healthy individuals (health group) in Nanjing Stomatological Hospital, Affiliated Hospital of Medical School, Nanjing University from June to October of 2022. Contents of kynurenine and the metabolites in saliva samples were analyzed by ultra-performance liquid chromatography-tandem mass spectrometry. The expressions of indoleamine 2, 3-dioxygenase (IDO) and aryl hydrocarbon receptor (AhR) were further detected by immunohistochemistry in gingival tissues. The PDLSC used in this study were isolated from extracted teeth for orthodontic treatment in Nanjing Stomatological Hospital, Affiliated Hospital of Medical School, Nanjing University from July to November of 2022. Experiments were then conducted using the cells by incubating with (kynurenine group) or without kynurenine (control group) in vitro. Seven days later, alkaline phosphatase (ALP) staining and assays of ALP activity were performed. Real-time fluorescence quantitative PCR (RT-qPCR) was utilized to detect the expressions of osteogenic related genes ALP, osteocalcin (OCN), runt-related transcription factor 2 (RUNX2), collagen type-Ⅰ (COL-Ⅰ) as well as the kynurenine pathway-associated genes AhR, cytochrome P450 family (CYP) 1A1, CYP1B1. Western blotting was used to detect the expression levels of RUNX2, osteopontin (OPN) and AhR proteins on day 10 and alizarin red staining was performed to observe the formation of mineral nodules on day 21 in control group and kynurenine group. Results: Salivary concentrations of kynurenine [8.26 (0, 19.60) nmol/L] and kynurenic acid [11.4 (3.34, 13.52) nmol/L] were significantly higher in the periodontitis group than in the health group [0.75(0, 4.25) nmol/L, 1.92(1.34, 3.88) nmol/L] (Z=-2.84, P=0.004; Z=-3.61, P<0.001). The expression levels of IDO (18.33±2.22) and AhR (44.14±13.63) in gingival tissues of periodontitis patients were significantly higher than that of the health group (12.21±2.87, 15.39±5.14) (t=3.38, P=0.015; t=3.42, P=0.027). In vitro, the ALP activity of PDLSC in the kynurenine group (291.90±2.35) decreased significantly compared with the control group (329.30±19.29) (t=3.34, P=0.029). The mRNA expression levels of ALP, OCN and RUNX2 in the kynurenine group (0.43±0.12, 0.78±0.09, 0.66±0.10) were decreased compared with the control group (1.02±0.22, 1.00±0.11, 1.00±0.01) (t=4.71, P=0.003; t=3.23, P=0.018; t=6.73, P<0.001), while the levels of AhR and CYP1A1 were increased in the kynurenine group (1.43±0.07, 1.65±0.10) compared with those in the control group (1.01±0.12, 1.01±0.14) (t=5.23, P=0.006; t=6.59, P<0.001). No significant difference was observed in COL-Ⅰ and CYP1B1 mRNA levels between groups. The protein levels of OPN, RUNX2 (0.82±0.05, 0.87±0.03) were reduced and that of AhR (1.24±0.14) was increased in the kynurenine group compared with those in the control group (1.00±0.00, 1.00±0.00, 1.00±0.00) (t=6.79, P=0.003; t=7.95, P=0.001; t=3.04, P=0.039). Conclusions: Over-activated kynurenine pathway in periodontitis patients can promote upregulation of AhR and suppress the osteogenic differentiation of PDLSC.

Analysis of the molecular typing and clinicopathological features of POLE and POLD1 gene mutations in middle-aged and elderly patients with colorectal cancer / 中华老年医学杂志

Objective:To examine the clinicopathological and molecular pathological characteristics of patients with colorectal cancer(CRC)who have mutations in the POLE and POLD1 genes.Methods:In this study, we retrospectively collected data from 276 middle-aged and elderly patients aged 45 years and over who were diagnosed with colorectal cancer at Beijing Hospital between October 2020 and September 2022.We utilized next generation sequencing and bioinformatics analysis to screen for harmful germline and somatic mutations in the POLE and POLD1 genes.The study involved 276 patients, who were divided into three groups based on their genetic mutations.The deleterious mutation group had 6 cases, the mutation of unknown significance group had 18 cases, and the wild type group had 252 cases.We also collected clinical and pathological features of the patients and analyzed their correlation with other molecular pathological results such as tumor mutation burden(TMB), microsatellite instability(MSI), and gene co-mutation.Results:No germline mutations were detected in the POLE and POLD1 genes across all patients.Out of the 276 patients, 18(6.5%)were found to carry POLE mutations.Among these, 6(2.2%)were classified as deleterious mutations, 12(4.3%)were positive for POLE mutations of unknown significance, and the remaining patients had wild type POLE genes.Out of the 276 patients, POLD1 gene mutations of unknown significance were found in 10 patients(3.6%). Among the 276 patients, 5 cases(1.8%)carried two types of gene mutations.Patients in the deleterious mutation group showed earlier tumor stage( P<0.05)and a higher prevalence of low-grade tumor budding( P<0.05), with 6 patients being affected by this.Compared to the wild type group, colon cancer patients showed a higher frequency of deleterious mutations and variants of unknown significance in the poorly differentiated group( P<0.05). The median TMB in the deleterious mutation group was 257.76 muts/Mb, 74.4 muts/Mb in the mutation of unknown significance group, and 5.81 muts/Mb in the wild type group.The study found significant differences in TMB-H status among the three groups, with all P-values less than 0.01.MSI-H status was detected in 1 case(16.7%, 1/6), 14 cases(77.8%, 14/18), and 18 cases(7.1%, 18/276)in the deleterious mutation group, variant of undetermined significance group, and wild type group, respectively.Notably, patients with variants of undetermined significance had a higher MSI-H status than patients with wild type and POLE deleterious mutations, with all P-values less than 0.01.The frequencies of co-mutations in KRAS, NRAS, BRAF, and PIK3CA were higher in the deleterious mutation group compared to the mutation of undetermined significance group and wild type group(all P<0.05). Conclusions:Colorectal cancer(CRC)patients with harmful mutations and variants of undetermined significance exhibit unique clinicopathological features.Patients with variants of undetermined significance are more likely to develop colon cancer, show poor differentiation, and have higher frequencies of TMB-H(tumor mutational burden)and MSI-H(microsatellite instability-high).

Ultrasound imaging and clinical characteristics of primary fallopian tube carcinoma in peri-and post-menopausal women / 中华老年医学杂志

Objective:To analyze the characteristic ultrasonographic findings and clinical symptoms of primary fallopian tube cancer(PFTC)in peri-and post-menopausal women, and to provide a basis for the early diagnosis of PFTC via ultrasonography.Methods:A total of 34 patients with PFTC confirmed by surgery and pathology in Beijing Hospital from May 2012 to January 2022 were retrospectively analyzed.Patients aged between 46-85 years, including 1(2.9%)in early menopausal transition(46 years old), 3(8.8%)in late menopausal transition(48 years old, 49 years old, and 50 years old), and 30(88.2%)in post-menopause.The median age was 60 years(60.9±9.7 years). The ultrasound and clinical features were analyzed and compared with pathological results.Results:Irregular vaginal bleeding was the most common clinical symptom in the 34 patients.According to the characteristics of adnexal masses and the involvement of other organs in the pelvis and abdominal cavity, the 34 cases of PFTC were divided into 4 groups.In Group A, 6 cases showed sausage-like cystic masses in the adnexal area, with unsmooth inner walls, medium echo papillary projections on the inner wall, incomplete strip partitions, and rich or relatively rich blood flow signals on the papillary projections and partitions.On ultrasound, 1 case was diagnosed with hydrosalpinx, 2 cases with cysts in the adnexal area, and 3 cases with PFTC.In Group B, 13 cases presented with cystic or solid masses of irregular shapes and sausage-like changes in some areas.The cystic part had poor sound transmission and the solid part exhibited rich or relatively rich blood flow signals.On ultrasound, 2 cases were diagnosed with PFTC and 11 cases with ovarian cancer with 2 showing intrauterine effusion and 3 showing abdominal and pelvic effusion.There were 11 cases in Group C, presenting with hypoechoic irregular solid masses in the adnexal area, with some accompanied by thickening of the peritoneum of the anterior and posterior fornix and the omentum.There were abundant or relatively abundant blood flow signals inside the masses.On ultrasound, 10 cases were diagnosed with ovarian cancer and 1 with pelvic metastatic cancer.There were 4 cases in Group D, who showed no adnexal mass, but 3 cases had ascites and were suspected of having a malignant tumor of unknown origin.Postoperative pathological results of the 34 PFTC cases revealed that 30 had high-grade serous carcinoma, 1 had low-grade serous carcinoma, 2 had serous tubal intraepithelial carcinoma(STIC), and 1 had serous tubal in-situ carcinoma.There were 3 cases with bilateral PFTC, 10 cases with PFTC on the left and 21 cases on the right.There were 4 cases with PFTC plus ovarian cancer.According to FIGO pathological staging, 8 cases were in stage Ⅰ, 9 in stage Ⅱ, 11 in stage Ⅲ and 6 in stage Ⅳ.The accuracy of preoperative diagnosis of PFTC by ultrasound was 50.0% in Group A and 15.4% in Group B. Preoperative ultrasound did not correctly diagnose PFTC in Group C and Group D, and instead suggested pelvic malignant tumor.Conclusions:Clear display of bilateral ovaries, sausage-like masses in the adnexal area, abundant or relatively abundant low resistive index blood flow signals in papillary projections and low-speed neovascularization are helpful ultrasound signs for the early diagnosis of PFTC, but not very useful for predicting the pathological stages of PFTC.Postmenopausal woman with vaginal bleeding, vaginal secretions or lower abdominal pain should be alert to PFTC.

Accelerating Magnetic Resonance Fingerprinting Using Hybrid Deep Learning and Iterative Reconstruction

Purpose@#To accelerate magnetic resonance fingerprinting (MRF) by developing a flexible deep learning reconstruction method. @*Materials and Methods@#Synthetic data were used to train a deep learning model. The trained model was then applied to MRF for different organs and diseases. Iterative reconstruction was performed outside the deep learning model, allowing a changeable encoding matrix, i.e., with flexibility of choice for image resolution, radiofrequency coil, k-space trajectory, and undersampling mask. In vivo experiments were performed on normal brain and prostate cancer volunteers to demonstrate the model performance and generalizability. @*Results@#In 400-dynamics brain MRF, direct nonuniform Fourier transform caused a slight increase of random fluctuations on the T2 map. These fluctuations were reduced with the proposed method. In prostate MRF, the proposed method suppressed fluctuations on both T1 and T2 maps. @*Conclusion@#The deep learning and iterative MRF reconstruction method described in this study was flexible with different acquisition settings such as radiofrequency coils. It is generalizable for different In vivo applications.

Phosphorylation of Cofilin-1 Enhances Paclitaxel Resistance of Epithelial Ovarian Cancer Cells by Inhibiting Apoptosis / 生物医学与环境科学（英文）

Objective@#To investigate the molecular mechanism of high phosphorylation levels of cofilin-1 (p-CFL-1) associated with paclitaxel resistance in epithelial ovarian cancer (EOC) cells.@*Methods@#Cells displaying varying levels of p-CFL-1 and CFL-1 were created by plasmid transfection and shRNA interference. Cell inhibition rate indicating paclitaxel efficacy was assessed by Cell Counting Kit-8 (CCK-8) assay. Apoptosis was assessed by flow cytometry and protein levels were detected by western blotting. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to measure the expression levels of phosphokinases and phosphatases of CFL-1. Survival analysis evaluated the correlation between the prognosis of EOC patients and the levels of p-CFL-1 and slingshot-1 (SSH-1).@*Results@#High levels of p-CFL-1 were observed in EOC cells that survived treatment with high doses of paclitaxel. SKOV3 cell mutants with upregulated p-CFL-1 showed impaired paclitaxel efficacy, as well as decreased apoptosis rates and pro-survival patterns of apoptosis-specific protein expression. Cytoplasmic accumulation of p-CFL-1 inhibited paclitaxel-induced mitochondrial apoptosis. SSH-1 silencing mediated CFL-1 phosphorylation in paclitaxel-resistant SKOV3 cells. Clinically, the high level of p-CFL-1 and the low level of SSH-1 in EOC tissues were closely related to chemotherapy resistance and poor prognosis in EOC patients.@*Conclusion@#The SSH-1/p-CFL-1 signaling pathway mediates paclitaxel resistance by apoptosis inhibition in EOC and is expected to be a potential prognostic predictor.

The co-relation of BRAF V600E mutation and factors affecting occurrence and prognosis of papillary thyroid carcinoma / 中华病理学杂志

Objective@#To investigate the prevalence of BRAF V600E mutation in thyroid nodules and to analyze the relationship between BRAF V600E mutation and various clinicopathological features.@*Methods@#BRAF V600E mutant gene test was done in 463 cases of thyroid nodules collected from April 2015 to July 2018 in Beijing Hospital. Pathologic sections of 444 cases of papillary thyroid carcinoma were reviewed and clinical information was collected.Statistical analysis of the relationship between BRAF V600E gene mutation and various clinicopathological features was performed with SPSS 21.0 statistical software.@*Results@#There were 109 males and 354 females in the cohort, with a male to female ratio of 1.0∶3.2. The patient ranged in age from 16 to 82 years, with an average age of 46.1 years. The BRAF V600E mutation rates in papillary thyroid carcinoma, benign thyroid nodules and other thyroid carcinoma were 86.5%(384/444),0/15 and 1/4,respectively.There was significant correlation between BRAF V600E mutation and histological diagnosis of papillary thyroid carcinoma (P<0.05). There was no correlation with age, gender, multifocality, bilaterality, coexisting lymphocytic thyroiditis, nodular goiter, maximum diameter, capsule invasion, extrathyroidal extension and clinical stage (P>0.05).@*Conclusions@#BRAF V600E gene mutation is closely related to the occurrence of papillary thyroid carcinoma. BRAF V600E has significant value in the diagnosis of papillary thyroid carcinoma. While BRAF V600E mutation is related to the histological diagnosis, it shows no correlation with other clinicopathologic features. BRAF V600E mutation is not an independent prognostic factor in papillary thyroid carcinoma patients.

Effects of Exercise Induced Autophagy on Antioxidant Defense Function of Skeletal Muscles in Mice / 中国运动医学杂志

Objective To explore whether a single bout of exhaustive exercise can activate autophagy in mice skeletal muscles and to investigate the effect and possible mechanisms of exercise-induced autophagy on antioxidant defense function in vivo.Methods Thirty healthy 4-week-old male C57BL/6 mice were randomly divided into a control group(n=6) and an exercise group(n=24).The latter was subjected to a single bout of exhaustive treadmill exercise.Gastrocnemius muscles of 6 mice on both sides were then isolated right after,as well as 6h,12h and 24h after the running.The manganese super oxide dismutase(MnSOD),total superoxide dismutase(T-SOD) and Copper-Zinc superoxide dismutase (CuznSOD) activities were determined using the xanthine oxidase method and the content of the total anti-oxygen capability(T-AOC) using the colorimetric method.The expression of Beclin1,P62 and Bcl2 mRNAs,as well as their proteins were determined using the fluorescent quantitative PCR and Western blotting.Results The expression of Beclin1 mRNA in gastrocnemius muscles of the exercise group increased more significantly than that of the control group right after the exercise,as well as 6 hours later(P＜0.01).P62 mRNA expression also increased significantly(P＜0.01) 24h after the exercise.In contrast,the expression of P62 protein at 0h,12h and 24h after the exercise decreased more significantly(P＜0.05 or P＜0.01) than that of the control group.Similarly,the expression of Bcl2 protein at 0h,6h and 12h increased significantly(P＜0.05 or P＜0.01).The MnSOD and CuznSOD activity increased significantly right after the exercise(P＜0.05).The T-SOD activity decreased significantly(P＜ 0.01) 6h after the exercise.T-AOC content increased significantly 6h,12h and 24h after the exercise (P＜0.05 or P＜0.01).The P62 protein expression was negatively correlated with T-AOC content.Conclusion The single bout of exhaustive treadmill exercise can activate the expression of such autophagyrelated factors as Beclin1 mRNA and P62 protein and increase the autophagy to a certain extent to maintain antioxidant defensive function of the skeletal muscle.

The level and influencing factors of nurses preparedness for disasters in nurses with low seniority in tertiary hospitals of Anhui province / 中国实用护理杂志

Objective To investigate the level of nurses′preparedness and influencing factors of nurses with low seniority in Anhui province. Methods Totally 295 nurses with low seniority were recruited by convenience sampling method. They were investigated by a self-designed demographic data questionnaire and the Disaster Preparedness Evaluation Tool. Results The score of the disaster nursing ability of the nursing managers was 163.85 ± 32.41, the mean score was 3.64 ± 0.72. Regression analysis showed that the gender, the experience of disaster rescue,disaster training were the influencing factors of disaster preparedness. Conclusions The capacity of disaster preparedness for nurses is not ideal. There is still room for improvement among nurses in Anhui province to enhance disaster preparedness, and improved disaster nursing training system is needed to achieve this.

Analysis on volume modulated arc therapy for spinal metastases / 中华放射医学与防护杂志

Objective To evaluate clinical effect of volume modulated arc therapy ( VMAT) for spinal metastases .Methods Twenty patients with spinal metastases who had been treated with VMAT were chosen to participate in the study .The prescribed dose varied from 45 to 60 Gy within 15 -20 fractions, 3 Gy per fraction, and one fraction per day of VMAT .Pain and quality of life were measured before VMAT and at several time points up to 6 months after treatment , by the numerical rating scale (NRS) and verbal rating scale (VRS) and the quality of life scale for bone metastases (EORTC QLQ-BM22) questionnaire.In addition, Frankel grading was used to evaluate the neurological function of spinal cord.The primary endpoint was frequency and duration of complete pain relief , and the secondary endpoint was death.Results At the end of the follow-up, the number of patients reporting no pain from spinal metastases, as measured by the NRS, increased from 0 of 20 before VMAT to 10 of 14 ( t =20.24, P0.05).After VMAT, the patients who suffered from nerve function impairment recovered to different extent . No one had radiation-induced injury , such as radiation myelitis , radiation pneumonitis , etc.Median survival time was 10 month.Conclusions VMAT is a safe and effective treatment method for spinal metastases .Significant reductions in patient-reported pain were observed , along with nerve dysfunction improved .The patients′quality of life was significantly improved .VMAT has no late spinal cord toxicities .

The relationship between phosphorylation at Ser3 of cofilin1 and Taxol resistance of ovarian cancer in elderly female patients / 中华老年医学杂志

Objectives To analyze the relationship between phosphorylation of serine-3 of cofilin1 and Taxol resistance of ovarian cancer and to evaluate the prognostic value of cofilin1 phosphorylation in estimating ovarian cancer survival using clinical samples.Methods Wild type (WT) plasmids with over-expression of wild cofilin1,S3L plasmids with over-expression of cofilin1 and inhibited phosphorylation at serine-3,and siRNA-C plasmids with down-regulation of cofilin1 were constructed using molecular biology techniques.After the SKOV3 and SK-TR30 cell-lines were transfected with these plasmids,changes in biological characteristics and drug resistance after instant transfection were compared.Fifty-one elderly female patients with microscopically confirmed ovarian cancer,who had received chemotherapy with Taxol and cis-platinum (TC)after surgery,including 30 chemo-sensitive and 21 chemo-resistant cases,were recruited in this study.Immunohistochemical methods were used to detect the expression of cofilinl and phosphorylated cofilin in tissue sections from the two groups.Progression free survival(PFS)was analyzed.Results Compared with the control group,the proportion of cells increased at the G0/G1 phase(P=0.034)and decreased at the S phase (P=0.031),and the apoptosis rate decreased(P=0.020),in SKOV3 cells with high expression of cofilin1.However,these characteristics disappeared when the wild type was replaced with the inactive mutant S3L.When cofilin1 expression was down-regulated in SK-TR30 cells,the proportion of cells at the G0/G1 phase decreased(P=0.020).The expression of cofilin1 was detected in 56.7 ％ (17/30)and 57.1％ (12/21),respectively,sections of ovarian tumor tissues of the chemo-sensitive and chemoresistant groups,and there was no statistic difference (x2=0.332,P =0.943)between the two groups.The expression of phosphorylated cofilin was much higher in the chemo-resistant group (85.7％ or 18/21)than in the chemo-sensitive group (53.3％ or 16/30),and the difference was statistically significant(x2=5.829,P=0.016).The higher expression of phosphorylated cofilin was also correlated with shorter PFS(x2 =21.440,P＜0.01,95％ CI:0.068-0.883),especially in the chemo-sensitive group.Conclusions Serine-3 phosphorylation status of cofilin 1 is associated with paclitaxel resistance in ovarian cancer,but the underlying mechanisms of regulation need further investigation.

Anatomic study and clinical practice of mesopancreas and total mesopancreatic excision / 中华外科杂志

Objective@#To explore the anatomical characteristics of the mesopancreas, to define the range of the total mesopancreas excision and to evaluate the feasibility, safety and effectiveness in the treatment of pancreatic cancer.@*Methods@#A regional anatomical and pathological study was performed on 14 cadavers with large slices and paraffin sections. The clinical and pathological data of 58 consecutive patients underwent total mesopancreas excision for pancreatic head carcinoma from January 2013 to December 2015 were prospectively collected and analysed. The perioperative morbidity, mortality and clinical outcomes of patients underwent total mesopancreas excision were compared with the patients underwent conventional pancreaticoduodenectomy from January 2010 to December 2012.@*Results@#The mesopancreas located in the retropancreatic area, extending from the head, neck, and uncinated process of pancreas to the aorto-caval groove, in which there were loose areolar tissue, adipose tissue, nerve plexus, lymphatic and capillaries. Although no fibrous sheath or fascia like mesocolorectum was found around the structures, a relatively fixed extent could be defined according to its embryologic and anatomic characters. In clinical practice, total mesopancreas excision was classified into two levels according to the extent of resection in this series: level Ⅰ was a"standard total mesopancreas excision" or"total mesopancreas excision in a narrow sense" , which was similar to the extent of standard resection from consensus statement of ISGPS. Level Ⅱ was defined as any procedure extending the range of level Ⅰ, called the"extended total mesopancreas excision" or"total mesopancreas excision in a broad sense". In TMpE group, the intraoperative blood loss( (461.4±184.5)ml vs. (532.2±319.8)ml, P=0.301), operation time( (368.6±92.5)minutes vs. (397.1±112.7)minutes, P=0.559), total complication rate (39.7% vs. 51.2%, P=0.250), fistula mortality (25.9% vs. 30.2%, P=0.628) were all reduced. There were significantly higher R0 rate (91.4% vs.76.7%, P=0.041) and more harvested lymph nodes (16.2 vs. 11.4, P=0.000) and lower total and local recurrence: rate (half-year local recurrence rate: 7.8% vs. 23.7%, P=0.036; one-year local recurrence rate: 18.2% vs. 39.5%, P=0.018) and longer disease-free survival (16.9 months vs. 13.4 months, P=0.044) and overall survival(22.5 months vs. 19.9 months, P>0.05) were also found in the study group.@*Conclusions@#Mesopancreas is different from mesorectum since it has no fascial envelop, which should be regarded as a surgical concept, rather than an anatomical structure. Total mesopancreas excision is safe and feasible for pancreatic head cancer and probably helps to increase the R0 resection rate and improve the clinical outcomes.

Accurate grading of pancreatic neuroendocrine tumors with Ki-67 index in fine-needle aspiration specimens: a comparative cytologic and histologic study / 中华病理学杂志

Objective@#To study the cytomorphologic features and determine whether pancreatic neuroendocrine tumors (PanNET) sampled by fine-needle aspiration (FNA) can be accurately graded based on the Ki-67 index when compared to surgical samples.@*Methods@#Corresponding intraoperative (19 cases) or endoscopic ultrasound-guided (3 cases) FNA cytology and surgical tissue specimens were obtained from 22 tumors, which were reviewed and stained for Ki-67 proliferation marker. The cytological samples included more than 200 tumor cells. Samples were graded by scoring the Ki-67 positive index in accordance with the 2010 WHO criteria. The grading scores assigned to the FNA cytology samples were compared with the scores assigned to the corresponding histological samples. Concordance was achieved by using 5% (instead of 2%) as a cut-off value for defining G2 tumors. One cytological sample included less than 500 tumor cells was excluded in the concordance calculation.@*Results@#The cytological smears consisted of uniform, monotonous and isolated cells, loose cellular aggregates and rosette-like formations. Some tumor cells clustered around segments of capillaries. The cells demonstrated distinct cytoplasmic and nuclear features. Mitoses and necrosis were rarely seen. When traditional 2% Ki-67 index cut-off value were used to classify G2 tumors, the majority (86.4%, κ=0.812, P<0.01) of FNA cytology samples and corresponding surgical tissue specimens demonstrated concordance. When a 5% cut-off value was adopted, the concordance rate was 95.5% (21/22, κ=1.000, P<0.01). Similar concordance rates between the cytological and histological grades were achieved with threshold value of cytological assessment material set at more than 500 or 200 cells.@*Conclusions@#The cytological Ki-67 index in adequate material (>200 tumor cells) is useful in grading pancreatic neuroendocrine tumors, and a cut-off value of 5% showed better predictive value compared with that of 2%. Accurate grading of PanNET is critical for predicting tumor biology, patient prognosis, and making informed decisions regarding patient management and treatment.

A new depsidone from stems-leaves of Garcinia multiflora / 中草药

Objective: To study the chemical constituents in stems-leaves of Garcinia multiflora. Methods: The chemical constituents from stems-leaves of G. multiflora were isolated by silica gel and HPLC methods. Their structures were elucidated by spectroscopic methods and physicochemical properties. Results: One new depsidone isolated from stems-leaves of G. multiflora was named as multidepsidone A (1), and four xanthones isolated from stems-leaves of G. multiflora were identified as isojacareubin (2), jacareubin (3), isocudraniaxanthone B (4), and 2-dimethylallyl-1,3,5-trihydroxyxanthone (5). Conclusion: Compound 1 is a new compound. compounds 2-4 are known compounds, and compounds 2 and 5 are obtained from this plant for the first time.

Assessment of two methods in evaluating alveolar bone loss by micro-CT based on periodontitis model in mice / 中国介入影像与治疗学

Objective To evaluate two methods measuring alveolar bone loss by micro computed tomography (micro-CT)based on periodontitis model in mice.Methods The silk ligatures were tied around the right maxillary second molars of mice to induce periodontitis model.The right half maxillaries of mice model were harvested for micro-CT analysis.Three dentists were recruited for the measurement with two different methods:Modified tomography (T) method and reconstruction (R) method.Accuracy and consistency of each method were estimated by standard deviation (SD).Results The SDs of R method managed by the same operator (measurement for 3 times) or different operators (3 operators) were 34.87μm and 35.67 μm respectively,while that of T method was 7.82 μm and 14.24 μm respectively.The SDs of T method were significantly lower than those of R method (both P＜0.05).Conclusion T method is more accurate and consistent than R method for evaluating alveolar bone loss in mice periodontitis model.

Correlation of chronic prostatic inflammation with prostate cancer and hyperplasia of prostate in the elderly / 中华老年医学杂志

Objective To investigate the pathological features of chronic inflammation in prostatic specimens,and study the correlation of chronic prostatic inflammation with prostate cancer and hyperplasia of prostate in the elderly.Methods The histopathologic features of prostatic specimens which were taken during prostatectomy were retrospectively observed.The inflammatory cells in partial cases were labeled by immunohistochemical markers such as CD3,CD20,CD4,CD8,CD117 and CD138,and by inflammation related factors such as nuclear factor(NF)-κB,(COX-2),tumor necrosis factor-α(TNF-α)and inducible nitric oxide synthase (iNOS).Results The chronic prostatic inflammation of different extent were found in 100 patients with prostate cancer and 76 patients with benign prostatic hyperplasia,among them 71 cases (40.3 ％) had mild chronic prostatic inflammation,80 cases (45.5 ％) had moderate chronic prostatic inflammation,25 cases (14.2 ％) had severe chronic prostatic inflammation.Inflammatory cells mainly were CD3-labeled lymphocytes,accompanied by a small amount of mononuclear cells and mast cells.Chronic prostatic inflammation was not correlated with prostatic carcinoma and its differentiation degree,benign prostatic hyperplasia,focal atrophy and its subtypes,and high grade prostatic intraepithelial neoplasia (all P＞ 0.05).There was correlation between chronic prostatic inflammation and expression of COX-2 (P＜ 0.05).Conclusions The histological chronic inflammation is common in the prostate specimens in elderly men.There are no direct correlations of prostatic inflammation with prostatic carcinomas and benign prostatic hyperplasia.Prostatic inflammation is positively correlated with the expression of COX 2,which may be associated with the activation of COX-2 pathway induced by oxidative stress.

A standardized protocol for detection of ALK protein expression and gene fusion in lung adenocarcinoma cytologic specimens / 中华肿瘤杂志

<p><b>OBJECTIVE</b>The aim of this study was to establish a standardized protocol for detection of ALK protein expression and gene fusion in cytologic specimens.</p><p><b>METHODS</b>Lung adenocarcinoma cytologic specimens were collected from seven hospitals in Beijing city. A detection protocol for ALK protein expression and gene fusion was designed according to the results of comparative experiment. Ventana immunohistochemical (IHC) ALK(D5F3) detecting ALK protein expression was performed in 203 prepared formalin-fixed paraffin-embedded (FFPE) cell blocks. ALK gene fusion in 98 EGFR gene wild type cytologic specimens and in 4 bronchoalveolar lavage fluid (BL) samples was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). ALK gene fusion in the Ventana IHC ALK (D5F3) positive samples was further tested by fluorescence in situ hybridization (FISH). Six patients with ALK IHC-positive result were followed up to analyze the responses of crizotinib therapy. Comparative experiments: (1) Comparison of the results of 4% neutral buffered formalin fixed for different time (24 h, 48 h, 72 h) on the Ventana IHC ALK (D5F3) staining was conducted in two cases of IHC ALK positive FFPE cell blocks; (2) Comparing qRT-PCR results for ALK fusion in samples from FFPE cell blocks and cytospin prepared slides in 10 cases of lung adenocarcinoma cytologic specimens.</p><p><b>RESULTS</b>Among the specimens examined using the standardized protocol recommended by this study, 229 cases of cytologic specimens met the diagnostic criteria of lung adenocarcinoma. Among them, 207 cases obtained ALK gene test results (by at least one method), with an ALK test ratio of 90.4% (207/229). FFPE cell blocks were successfully prepared in 203 cases, Ventana IHC ALK (D5F3) were successfully performed in all the 203 FFPE cell blocks (100%), and the ALK protein positive detection rate was 10.3% (21/203). ALK fusion was tested in 98 FFPE cytologic samples of EGFR wild types by qRT-PCR, and 96 out of 98 (97.96%) cytologic samples were successfully performed.18 out of 19 IHC ALK-positive cases were verified to be of ALK fusion status by qRT-PCR. The concordance rate was 94.7% (Kappa=0.967, P<0.001) between Ventana IHC ALK (D5F3) and qRT-PCR, and the sensitivity of the Ventana IHC ALK (D5F3) assay compared with qRT-PCR was 100% and the specificity was 98.7%. FISH assay was used to verify the positive cases detected by Ventana IHC ALK (D5F3) staining. Two cases of low tumor cell content FFPE samples obtained indefinite results by FISH test. The six patients with positive ALK protein expression received crizotinib therapy, and 5 paitents got treated effectively. For two ALK IHC positive cases, which were 4% neutral buffered formalin fixed for 72 h, the result of Ventana IHC ALK (D5F3) staining became weakened obviously and uneven. In 10 cases of samples, total RNA was extracted from FFPE cytologic sections and cytospin prepared slides, and the results of qRT-PCR test and ALK gene fusion showed good concordance.</p><p><b>CONCLUSIONS</b>The standardized protocol recommended in this study expands the detection types and quantity of cytologic specimens for ALK protein expression and gene fusion and increased the detection rate. Ventana IHC ALK (D5F3) is a reliable method for detecting ALK protein expression in FFPE cell blocks. The pathologic quality control procedure prior to Ventana IHC ALK (D5F3) is crucial for the accuracy of testing the ALK gene status. When FFPE cell blocks could not be prepared or prepared unsuccessfully from the cytologic specimens, qRT-PCR may be an alternative option for the detection of ALK gene fusion.</p>

Expression of Girdin in brain tissues of Alzheimer's disease / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the expression of Girdin and its significance in Alzheimer's disease (AD).</p><p><b>METHODS</b>Fifty-nine autopsy cases from Department of Pathology, Beijing Hospital from January 1988 to December 2013, including 35 AD cases and 24 non-AD cases as control. Girdin and amyloid β-protein (Aβ) expression was evaluated immunohistochemically by EnVision method. The correlation between Girdin and Aβ was analyzed.</p><p><b>RESULTS</b>Girdin expression was localized in the nucleus and/or cytoplasm. The expression rates of Girdin were 20.0% (7/35) in the AD group and 83.3% (20/24) in the non-AD group, respectively. The difference was significant (Yates's correction for continuity χ(2)=20.527, P<0.05). Girdin expression and Aβ deposition also correlated significantly (P<0.05).</p><p><b>CONCLUSIONS</b>Girdin shows reduced expression in AD, and is correlated positively with Aβ deposition. This suggests that Girdin may play an important role in the occurrence and development of AD.</p>

Progress on development and research of human bocavirus 1 / 病毒学报

Human bocavirus 1 (HBoV1) is a novel virus that mainly causes respiratory tract infection, and it has the characteristic of genome of Parvovirus, containing three open reading frames that encode non-structural proteins NS1 and NP1 and structural proteins VP1 and VP2. Circular episome is present during the rolling circle replication of HBoV1, which provides the possibility of full genome amplification and infectious clone construction to save HBoV1. The recombination between HBoV1 and HBoV2-4 occurs frequently. With the three-dimensional culture, in vitro culture of HBoV1 provides a powerful tool for research on the pathogenesis of HBoV1. This review focuses on the molecular characteristics, association with diseases, in vitro culture, diagnosis and treatment of HBoV1.

Genome amplification and sequence analysis of human bocavirus 2 / 病毒学报

To obtain the genome sequence of human bocavirus 2 (HBoV2), different regions of HBoV2 genome were amplified through PCR in fecal specimens which had been identified as single-positive for HBoV2 in 2010. A genome sequence of HBoV2 (HBoV2-NC, 5444 bp) was obtained after sequence assembly. The phylogenetic analysis showed that HBoV2-NC had the closest evolutionary relationship with HBoV2 Lanzhou strain. The predication of inverted terminal repeats of HBoV2-NC by DINAMelt showed that inverted terminal repeats were contained in HBoV2-NC 5' terminal, which had the typical stem-loop structure in other parvoviruses. Finally, some flanking sequences of HBoV2-NC were amplified by linker-PCR.

Clinical application of stereotactic body radiation therapy for colorectal cancer with liver metastases / 中华放射医学与防护杂志

Objective To evaluate the efficacy of stereotactic body radiation therapy (SBRT) for hepatic metastases from colorectal cancer,and to collect data for the application of this technique.Methods A total of 28 patients from No.306 Hospital of PLA,including 17 male and 11 female with median age of 63.8 (range from 31 to 86),were treated with SBRT for colorectal cancer with liver metastases with 54 lesions in total.The GTV,CTV and PTV were delineated above the enhanced CT scans acquired during normal quiet respiration.CTV was obtained by adding 5 mm isotropic margin from GTV,and PTV was obtained by adding 5 to 10 mm isotropic margin from CTV.Prescription dose line covered 50％-60％ of isodose curve at 3-6 Gy/fraction.The total dose was 39-45 Gy and the biologically equivalent doses(BED)was 50.7-65.3 Gy.The patients were followed-up beginning at 3 months after SBRT.The change in size of the lesion based on enhanced CT or MR scans was evaluated.Toxicity was evaluated and scored according to the RTOG criteria.Local control rate and survival rate were analysed.Results All patients completed the treatment.With median follow-up of 15.1 months (range frome 3 to 30 months),7 patients survived at the end of follow-up.The local control rate (LC) was 79.2％,and 1-and 2-year overall survival rate(OS) were 82.7％ and 48.6％,respectively.There was a close corelation between the size of lesion and the LC.The LC (PR + CR) was much better at the size of lesion less than 14 cm3 than that at the size more than 65 cm3(x2 =4.17,P＜0.05).When the size was more than 180 cm3,the LC was zero.Toxicity included fatigue (60.7％),grade 1 and 2 digestive system toxicity (28.6％),a transient grade 1 and 2 bone marrow suppression (46.4％) and a transient increase in transaminase(17.8％).No grade 3 toxicity and above and late toxicity were observed.Conclusions Stereotactic body radiation therapy could be suggested as the first choice for the selected patients who suffer form colorectal liver metastases,especially for those who cannot undergo surgery.