[Diagnosis and differential diagnosis of large B-cell lymphoma with IRF4 rearrangement].

Objective: To analyze the clinicopathological features and differential diagnosis of large B-cell lymphoma with IRF4 rearrangement, aiming enhance its recognition and prevent misdiagnosis. Methods: The clinicopathological features, immunophenotype, and fluorescence in situ hybridization (FISH) results of six cases diagnosed with IRF4 rearrangement-positive B-cell lymphoma at the Affiliated Hospital of Xuzhou Medical University from 2015 to 2023 were retrospectively analyzed. Additionally, a comprehensive review of the literature was conducted. Results: Six patients with IRF4 rearrangement-positive large B-cell lymphoma were included. Patients 1 to 5 included three males and two females with a median age of 19 years ranging from 11 to 34 years. Four patients presented with head and neck lesions, while the other one had a breast nodule; all were in clinical Ann Arbor stages I to Ⅱ. Morphologically, entirely diffuse pattern was present in two cases, purely follicular pattern in one case, and diffuse and follicular patterns in other two cases. The tumor cells, predominantly centroblasts mixed with some irregular centrocytes, were of medium to large size, with a starry sky appearance observed in two cases. Immunophenotyping revealed all cases were positive for bcl-6 and MUM1, with a Ki-67 index ranging from 70% to 90%, and CD10 was positive in two cases. IRF4 rearrangement was confirmed in all cases by FISH analysis, with dual IRF4/bcl-6 rearrangements identified in two cases, leading to a diagnosis of LBCL-IRF4. Case 6, a 39-year-old female with a tonsillar mass and classified as clinical Ann Arbor stage Ⅳ, displayed predominantly diffuse large B-cell lymphoma (DLBCL) morphology with 20% high-grade follicular lymphoma characteristics. Immunohistochemistry showed negative CD10 and positive bcl-6/MUM1, with a Ki-67 index of approximately 80%. Triple rearrangements of IRF4/bcl-2/bcl-6 were identified by FISH, leading to a diagnosis of DLBCL with 20% follicular lymphoma (FL). All six patients achieved complete remission after treatment, with no progression or relapse during a follow-up period of 31-100 months. Conclusions: Large B-cell lymphoma with IRF4 rearrangement is a rare entity with pathological features that overlap with those of FL and DLBCL. While IRF4 rearrangement is necessary for diagnosing LBCL-IRF4, it is not specific and requires differentiation from other aggressive B-cell lymphomas with IRF4 rearrangement.

Agrobacterium-delivered VirE2 interacts with host nucleoporin CG1 to facilitate the nuclear import of VirE2-coated T complex.

Agrobacterium tumefaciens is the causal agent of crown gall disease. The bacterium is capable of transferring a segment of single-stranded DNA (ssDNA) into recipient cells during the transformation process, and it has been widely used as a genetic modification tool for plants and nonplant organisms. Transferred DNA (T-DNA) has been proposed to be escorted by two virulence proteins, VirD2 and VirE2, as a nucleoprotein complex (T-complex) that targets the host nucleus. However, it is not clear how such a proposed large DNA-protein complex is delivered through the host nuclear pore in a natural setting. Here, we studied the natural nuclear import of the Agrobacterium-delivered ssDNA-binding protein VirE2 inside plant cells by using a split-GFP approach with a newly constructed T-DNA-free strain. Our results demonstrate that VirE2 is targeted into the host nucleus in a VirD2- and T-DNA-dependent manner. In contrast with VirD2 that binds to plant importin α for nuclear import, VirE2 directly interacts with the host nuclear pore complex component nucleoporin CG1 to facilitate its nuclear uptake and the transformation process. Our data suggest a cooperative nuclear import model in which T-DNA is guided to the host nuclear pore by VirD2 and passes through the pore with the assistance of interactions between VirE2 and host nucleoporin CG1. We hypothesize that this large linear nucleoprotein complex (T-complex) is targeted to the nucleus by a "head" guide from the VirD2-importin interaction and into the nucleus by a lateral assistance from the VirE2-nucleoporin interaction.

[Mucinous tubular and spindle cell carcinoma of kidney: Clinicopathology and prognosis].

OBJECTIVE: To investigate and summarize the clinicopathological features, immunophenotype, differential diagnosis and prognosis analysis of mucinous tubular and spindle cell carcinoma (MTSCC). METHODS: The data of thirteen cases of MTSCC were retrospectively analyzed, the clinical and pathological characteristics and immunohistochemical expression were summarized, and fluorescence in situ hybridization was detected. RESULTS: Among the thirteen patients, four were males and nine females, with a male-to-female ratio of 1 ∶2.25. The average age was 57.1 years, ranging from 39 to 78 years. The maximum diameter of the tumor was 2-12 cm. All cases had no symptoms, and were accidentally discovered, 3 cases underwent partial renal resection, 10 cases underwent radical renal resection, 9 cases were located in the left kidney, and 4 cases were located in the right kidney. Most of the cases showed the classical morphological changes, with 11 cases of nuclear grading [World Health Organization (WHO)/International Society of Urological Pathology (ISUP) grading system] being G2 and 2 cases being G3. There were 6 cases of stage PT1a, 3 cases of PT1b, 2 cases of PT2a, and 1 case of PT2b and 1 case of PT3a. The positive rates of immunohistochemical staining were: vimentin, AE1/AE3, α-methylacyl-CoA racemase (αMACR) and cytokeratin (CK) 8/18, 100% (13/13); CK7, 92.3% (12/13); epithelial membrane antigen (EMA), 92.3% (12/13); CK20, 46.2% (6/13); CD10, 30.8% (4/13); synaptophysin (Syn), 7.7% (1/13); chromogranin A (CgA), CD57, WT1 and Ki-67, 0 (0/13), and fluorescence in situ hybridization showed that no trisomy of chromosomes 7 and 17 were observed in any of the cases. The follow-up period was 6 months to 7 years and 6 months, 2 cases died after lung metastasis (one with ISUP/WHO grade G3, one with necrosis), and the remaining 11 cases had no recurrence and metastasis. CONCLUSION: MTSCC is a unique type of low-grade malignancy kidney tumor, occurs predominantly in females, widely distributed in age, the current treatment method is surgical resection, and cases with necrosis and high-grade morphology are prone to recurrence and metastasis, although most cases have a good prognosis, but they still need close follow-up after surgery.

[Impact of different diagnostic criteria for assessing mild micro-hepatic encephalopathy in liver cirrhosis: an analysis based on a prospective, multicenter, real-world study].

Objective: To compare the differences in the prevalence of mild micro-hepatic encephalopathy (MHE) among patients with cirrhosis by using the psychometric hepatic encephalopathy score (PHES) and the Stroop smartphone application (Encephal App) test. Methods: This prospective, multi-center, real-world study was initiated by the National Clinical Medical Research Center for Infectious Diseases and the Portal Hypertension Alliance and registered with International ClinicalTrials.gov (NCT05140837). 354 cases of cirrhosis were enrolled in 19 hospitals across the country. PHES (including digital connection tests A and B, digital symbol tests, trajectory drawing tests, and serial management tests) and the Stroop test were conducted in all of them. PHES was differentiated using standard diagnostic criteria established by the two studies in China and South Korea. The Stroop test was evaluated based on the criteria of the research and development team. The impact of different diagnostic standards or methods on the incidence of MHE in patients with cirrhosis was analyzed. Data between groups were differentiated using the t-test, Mann-Whitney U test, and χ (2) test. A kappa test was used to compare the consistency between groups. Results: After PHES, the prevalence of MHE among 354 cases of cirrhosis was 78.53% and 15.25%, respectively, based on Chinese research standards and Korean research normal value standards. However, the prevalence of MHE was 56.78% based on the Stroop test, and the differences in pairwise comparisons among the three groups were statistically significant (kappa = -0.064, P < 0.001). Stratified analysis revealed that the MHE prevalence in three groups of patients with Child-Pugh classes A, B, and C was 74.14%, 83.33%, and 88.24%, respectively, according to the normal value standards of Chinese researchers, while the MHE prevalence rates in three groups of patients with Child-Pugh classes A, B, and C were 8.29%, 23.53%, and 38.24%, respectively, according to the normal value standards of Korean researchers. Furthermore, the prevalence rates of MHE in the three groups of patients with Child-Pugh grades A, B, and C were 52.68%, 58.82%, and 73.53%, respectively, according to the Stroop test standard. However, among the results of each diagnostic standard, the prevalence of MHE showed an increasing trend with an increasing Child-Pugh grade. Further comparison demonstrated that the scores obtained by the number connection test A and the number symbol test were consistent according to the normal value standards of the two studies in China and South Korea (Z = -0.982, -1.702; P = 0.326, 0.089), while the other three sub-tests had significant differences (P < 0.001). Conclusion: The prevalence rate of MHE in the cirrhotic population is high, but the prevalence of MHE obtained by using different diagnostic criteria or methods varies greatly. Therefore, in line with the current changes in demographics and disease spectrum, it is necessary to enroll a larger sample size of a healthy population as a control. Moreover, the establishment of more reliable diagnostic scoring criteria will serve as a basis for obtaining accurate MHE incidence and formulating diagnosis and treatment strategies in cirrhotic populations.

[Identification of metabolic biomarkers associated with the onset of type 2 diabetes based on a nested case-control study].

Objective: To explore metabolic biomarkers associated with the onset of type 2 diabetes. Methods: Cluster random sampling method was used to select 10 867 local residents aged ≥ 20 years in Liangxi district of Wuxi City, Jiangsu Province in 2007. The baseline survey and physical examination were conducted to collect participants' information, including demographic characteristics, behavior and lifestyles, disease history, family history of diabetes, height, weight, waist circumference and blood pressure, etc. Blood samples were collected and biochemical indexes (high density lipoprotein cholesterol, total cholesterol, triglyceride, fasting blood glucose, etc.) were tested. By June 30, 2020, 220 newly diagnosed patients with type 2 diabetes during the follow-up were selected as cases, and 220 healthy individuals were matched as controls with age (±5 years) and the same sex. High performance liquid chromatography mass spectrometer was used to detect and identify metabolites in serum samples of two groups at baseline. Lasso regression and multivariate conditional logistic regression were used to explore the metabolites associated with the onset of type 2 diabetes. Results: The age of participants at baseline was (53±7) years, and 41.82% were male. 25 out of 1 579 metabolites were selected to be potentially associated with the onset of type 2 diabetes in the lasso regression model. The multivariable conditional logistic regression analysis showed that only 7-Methylxanthine had an independent effect on type 2 diabetes (P=0.019). The area under the receiver operating characteristic curve (AUC) (95%CI) of the prediction model of type 2 diabetes based on traditional risk factors was 0.80 (0.76-0.85). After the 7-methylxanthine in the model, the AUC (95%CI) increased to 0.92 (0.89-0.95) (P<0.001). From the second year, 7-methylxanthine could improve the prediction performance (P=0.007). Conclusion: The level of 7-methylxanthine is related to the onset of type 2 diabetes, and can be used as a biomarker to predict its incidence risk.

[Efficacy and safety of percutaneous transluminal pulmonary angioplasty in patients with chronic thromboembolic pulmonary hypertension].

Objective: To explore the efficacy and safety of percutaneous transluminal pulmonary angioplasty (PTPA) in patients with chronic thromboembolic pulmonary hypertension (CTEPH). Methods: This prospective single arm study included 19 CTEPH patients (7 male, age(56.3±12.5)years) admitted to Wuhan Asia Heart Hospital from January 2017 to June 2019 and received PTPA interventional therapy. Baseline data, including age, sex, WHO functional class, 6-minute walk distance (6MWD), NT-proBNP, right heart catheterization values, were collected. Patients received single or repeated PTPA. Number of dilated vessels from each patient was analyzed, patients were followed up for 24 weeks and right heart catheterization was repeated at 24 weeks post initial PTPA. All-cause death, perioperative complications, and reperfusion pulmonary edema were reported. WHO functional class, 6MWD, NT-proBNP, right heart catheterization values were compared between baseline and at 24 weeks follow up. Results: Nineteen CTEPH patients received a total of 56 PTPA treatments. The pulmonary artery pressure (mPAP) decreased from (40.11±7.55) mmHg (1 mmHg=0.133 kPa) to (27.53±4.75) mmHg (P<0.001), and the total pulmonary resistance (TPR) decreased from (13.00±3.56) Wood U to (5.48±1.56) Wood U (P<0.001), cardiac output increased from (3.19±0.63) L/min to (5.23±0.94) L/minutes (P<0.01) at 24 weeks post PTPA. The WHO functional class improved significantly (P<0.001), 6MWD increased from (307.08±129.51) m to (428.00±112.64) m (P=0.002), the NT-proBNP decreased at 24 weeks post PTPA (P=0.002). During the follow-up period, there was no death; hemoptysis occurred in 4 patients during the operation, none of which resulted in serious adverse clinical consequences. One patient developed reperfusion pulmonary edema and recovered after treatment. Conclusion: PTPA treatment is safe and can significantly improve the hemodynamics and WHO functional class of patients with CTEPH.

[Analysis of lung cancer screening through low-dose spiral computed tomography among petroleum company staffs in Sichuan-Chongqing Area in 2020].

Objective: To provide scientific evidence for early lung cancer screening, to analyze the incidence of pulmonary nodules among petroleum company staffs in Sichuan-Chongqing Area. Methods: In January 2021 , 6002 petroleum company staffs in Sichuan-Chongqing Area which scanned by low-dose spiral computed tomography (LDCT) of chest in medical examination center in 2020 were retrospectively collected as objects. Their imaging and clinical data were collected. χ(2) test was used to analyze the differences in the detection rates of lung nodules and suspected lung cancer nodules among workers in petroleum company staffs of different genders, ages and types of work. Results: Among the 6002 objects, 3853 (64.2%) were male and 2149 (35.8%) were female, with an average age of (47.25±12.13) years old. A total of 431 cases (7.2%) of pulmonary nodules and 57 cases (0.9%) of suspected lung cancer nodules were detected. 45 cases were followed up with surgical treatment, and 41 cases (91.1%) of lung cancer were diagnosed by postoperative pathology. There were significant differences in the detection rates of pulmonary nodules and suspected lung cancer nodules between different age groups (χ(2)=51.23, 18.81 , P<0.001). The detection rates of pulmonary nodules in the age groups 51-60 years old and ≥61 years old were higher than those in the age groups≤40 years old and 41-50 years old (P<0.05). The detection rate of suspected lung cancer nodules in the age group≥ 61 years old was higher than those in the age groups≤40 years old, 41-50 years old and 51-60 years old (P< 0.05) . And the detection rate of suspected lung cancer pulmonary nodules in oil workers was higher than that of ordinary workers (P<0.05) . Among female objects, the detection rate of pulmonary nodules in oil workers was higher than that in ordinary workers (χ(2)=8.09, P=0.004) . The detection rate of pulmonary nodules in oil workers aged ≥61 years old was higher than ordinary workers (χ(2)=37.94, P<0.001) . Among male objects, the detection rate of suspected lung cancer pulmonary nodules in oil workers was higher than that in ordinary workers (χ(2)=8.42, P=0.004) . The detection rates of suspected lung cancer pulmonary nodules in oil workers aged 51-60 years old and ≥61 years old groups were higher than those of ordinary workers (χ(2)=4.70, 8.74; P=0.030, 0.003) . Conclusion: LDCT is suitable for early lung cancer screening for petroleum company staffs. During the clinical screening process, LDCT should be used as a routine physical examination item for petroleum workers older than 51 years old.

[Clinicopathological features and prognosis of fumarate hydratase deficient renal cell carcinoma].

OBJECTIVE: To investigate the clinicopathological features and prognosis of fumarate hydratase deficient renal cell carcinoma (FH-RCC). METHODS: Immunohistochemical (IHC) staining was used to detect the expression of fumarate hydratase (FH) in tumor tissues of 109 different types of renal cell carcinoma (RCC) patients aged 60 years and younger from the Department of Urology of Peking University First Hospital from January 2013 to December 2019. The clinicopathological data and prognosis of FH-RCC were collected and analyzed. RESULTS: There were eleven patients with FH-negative expression. Seven were males and four females. The age of onset ranged 16-53 years (mean age: 36.7 years), and four female patients all had a history of uterine leiomyoma. Only one first-degree relative of one patient had renal cancer, and none of the patients had a history or family history of cutaneous leiomyomas. The diameter of the tumor was 2.1-12.0 cm (mean: 8.83 cm). Renal sinus or perirenal fat invasion was seen in nine cases, tumor thrombus in renal vein or inferior vena cava in six cases, lymph node metastasis in seven cases, adrenal gland invasion in four cases and splenic capsule invasion in one case. The cases were initially diagnosed as type Ⅱ papillary RCC (7/49, 14.3%), collecting duct carcinoma (2/9, 22.2%) and unclassified RCC (2/51, 3.9%). Tumor histopathology mostly showed a mixture of different structures, such as papillary, tubular cystic, solid, and so on. The most common histological structures were papillary (9/11, 81.8%) and tubular (8/11, 72.7%). Three cases had sarcomatoid areas. At least focal eosinophilic nucleolus (WHO/grades Ⅲ-Ⅳ) and perinuclear halo could be seen in all cases. Immunohistochemical (IHC) stains of most tumors were negative for CA9, CD10 and CK7. The results of fluorescence in situ hybridization (FISH) showed that there was no translocation or amplification of TFE3 gene in two cases with TFE3 IHC expression. All the patients were followed up for 11-82 months. Mean survival was 24 months. Five cases died of distant metastasis 9-31 months after operation (mean: 19 months), and five of the six patients alive had became metastatic. CONCLUSION: Morphologically, FH-RCC overlaps with many types cell RCC. A mixture of papillary and tubular cystic arrangement is the most common growth pattern of FH-RCC. At least focally large and obvious eosinophilic nucleoli are an important histological feature of this tumor. The negative expression of FH can help to confirm the diagnosis. Young female RCC patients with uterine leiomyomas should be suspected of FH-RCC. Some FH-RCC cases lack clinical evidence. The suspicion raised by pathologists based on histological characteristics is often the key step to further genetic testing and the final diagnosis of the tumor.

[Feasibility multi-center study of artificial intelligence assistance in cervical fluid-based cytology diagnosis].

Objective: To propose a method of cervical cytology screening based on deep convolutional neural network and compare it with the diagnosis of cytologists. Method: The deep segmentation network was used to extract 618 333 regions of interest (ROI) from 5, 516 cytological pathological images. Combined with the experience of physicians, the deep classification network with the ability to analyze ROI was trained. The classification results were used to construct features, and the decision model was used to complete the classification of cytopathological images. Results: The sensitivity and specificity were 89.72%, 58.48%, 33.95% and 95.94% respectively. Among the smears derived from four different preparation methods, this algorithm had the best effect on natural fallout with a sensitivity of 91.10%, specificity of 69.32%, positive predictive rate of 41.41%, and negative predictive rate of 97.03%. Conclusion: Deep convolutional neural network image recognition technology can be applied to cervical cytology screening.

Clinical characteristics of COVID-19-infected cancer patients: a retrospective case study in three hospitals within Wuhan, China.

BACKGROUND: Cancer patients are regarded as a highly vulnerable group in the current Coronavirus Disease 2019 (COVID-19) pandemic. To date, the clinical characteristics of COVID-19-infected cancer patients remain largely unknown. PATIENTS AND METHODS: In this retrospective cohort study, we included cancer patients with laboratory-confirmed COVID-19 from three designated hospitals in Wuhan, China. Clinical data were collected from medical records from 13 January 2020 to 26 February 2020. Univariate and multivariate analyses were carried out to assess the risk factors associated with severe events defined as a condition requiring admission to an intensive care unit, the use of mechanical ventilation, or death. RESULTS: A total of 28 COVID-19-infected cancer patients were included; 17 (60.7%) patients were male. Median (interquartile range) age was 65.0 (56.0-70.0) years. Lung cancer was the most frequent cancer type (n = 7; 25.0%). Eight (28.6%) patients were suspected to have hospital-associated transmission. The following clinical features were shown in our cohort: fever (n = 23, 82.1%), dry cough (n = 22, 81%), and dyspnoea (n = 14, 50.0%), along with lymphopaenia (n = 23, 82.1%), high level of high-sensitivity C-reactive protein (n = 23, 82.1%), anaemia (n = 21, 75.0%), and hypoproteinaemia (n = 25, 89.3%). The common chest computed tomography (CT) findings were ground-glass opacity (n = 21, 75.0%) and patchy consolidation (n = 13, 46.3%). A total of 15 (53.6%) patients had severe events and the mortality rate was 28.6%. If the last antitumour treatment was within 14 days, it significantly increased the risk of developing severe events [hazard ratio (HR) = 4.079, 95% confidence interval (CI) 1.086-15.322, P = 0.037]. Furthermore, patchy consolidation on CT on admission was associated with a higher risk of developing severe events (HR = 5.438, 95% CI 1.498-19.748, P = 0.010). CONCLUSIONS: Cancer patients show deteriorating conditions and poor outcomes from the COVID-19 infection. It is recommended that cancer patients receiving antitumour treatments should have vigorous screening for COVID-19 infection and should avoid treatments causing immunosuppression or have their dosages decreased in case of COVID-19 coinfection.

The clinical significance of MIA gene in tumorigenesis of lung cancer.

Lung cancer is a common malignant disease in humans. Both the incidence rate and death rate keep growing in recent years and the prognosis of lung cancer patients is disappointing. Melanoma inhibitory activity (MIA) is a secreted protein and a serum marker for metastasis of melanoma. MIA was reported as an oncogene in several cancers. But its role in lung cancer was unknown. In this study, MIA level was shown to be increased in peripheral blood of 216 patients with lung cancer. And it was expressed much higher in tumor tissues than the normal control. Moreover, MIA expression was associated with the clinical stage of lung cancer. When MIA was knocked down, the viability, migration and invasion of A549 cells were remarkably suppressed. But the cell apoptosis rate was enhanced reversely. In contrast, overexpression of MIA promoted cell proliferation, migration and invasion while cell apoptosis was inhibited. Mechanically, the anti-apoptosis marker Bcl-2 was increased and pro-apoptosis marker Bax was decreased after MIA was overexpressed in A549 cells, and vice versa. The level of PCNA and PI3K/mTOR signaling molecules was also increased when MIA was upregulated but declined after knockdown of MIA. In conclusion, MIA plays an oncogenic role in lung cancer and might be a potential marker for the diagnosis of lung cancer.

Agrobacterium-delivered virulence protein VirE2 is trafficked inside host cells via a myosin XI-K-powered ER/actin network.

Agrobacterium tumefaciens causes crown gall tumors on various plants by delivering transferred DNA (T-DNA) and virulence proteins into host plant cells. Under laboratory conditions, the bacterium is widely used as a vector to genetically modify a wide range of organisms, including plants, yeasts, fungi, and algae. Various studies suggest that T-DNA is protected inside host cells by VirE2, one of the virulence proteins. However, it is not clear how Agrobacterium-delivered factors are trafficked through the cytoplasm. In this study, we monitored the movement of Agrobacterium-delivered VirE2 inside plant cells by using a split-GFP approach in real time. Agrobacterium-delivered VirE2 trafficked via the endoplasmic reticulum (ER) and F-actin network inside plant cells. During this process, VirE2 was aggregated as filamentous structures and was present on the cytosolic side of the ER. VirE2 movement was powered by myosin XI-K. Thus, exogenously produced and delivered VirE2 protein can use the endogenous host ER/actin network for movement inside host cells. The A. tumefaciens pathogen hijacks the conserved host infrastructure for virulence trafficking. Well-conserved infrastructure may be useful for Agrobacterium to target a wide range of recipient cells and achieve a high efficiency of transformation.

[A clinical analysis of 14 cases of prostatic stromal tumor of uncertain malignant potential].

OBJECTIVE: To investigate the clinical and pathologic characteristics, diagnosis, treatment, prognosis and survival of prostatic stromal tumor of uncertain malignant potential. METHODS: Overall 14 patients with prostatic stromal tumor of uncertain malignant potential were treated from October 2008 to April 2020, the patient age ranged from 27 to 78 years (mean 54 years). The disease duration was 1 to 180 months (mean duration of 46 months). The clinical manifestations mainly included urinary obstructive symptoms and urethral irritating symptoms. The tumors were located in the peripheral zone or the transition zone. Digital rectum examination indicated prostatic tumor. Serum prostatic specific antigen level was always normal or elevated. Transrectal ultrasonography and magnetic resonance imaging indicated prostatic tumor. Magnetic resonance imaging in showed large, round, well-defined masses, which were diffusely heterogeneous signal on T2 weighted imaging. Following the administration of intravenous contrast medium, the lesion had diffuse and heterogeneous enhancement. RESULTS: In the study, 3 cases underwent prostate biopsy, 2 cases underwent transurethral resection of the prostate, 9 cases underwent radical excision or transurethral resection of the prostate with definite diagnosis of pathologic features. Under the light microscope, the interstitial cells of stromal tumor of uncertain malignant potential were overgrowth and fusiform cells showed some degree of pleomorphism, nuclei with few mitotic figures, and necrosis was not often seen. Immunohistochemical staining showed that prostate specific antigen was negative, while vimentin was positive in the tumor tissue, CD34, progesterone receptor and smooth muscle actin were positive in the majority, and Ki67 positive index was 1%-20% (mean 6%). Twelve cases were followed-up, and the time of survival varied from 10 to 96 months (mean 65 months), two cases were lost to the follow-up, one case died of disease at the end of 10 months, nine cases were free of disease recurrence after surgery, two cases underwent more transurethral resection of the prostate due to local recurrence. CONCLUSION: STUMP is a very rare tumor of the specialized prostatic stroma with an unpredictable clinical behavior. The clinical manifestations, transrectal ultrasonography and magnetic resonance imaging are valuable for the diagnosis of prostatic stromal tumor of uncertain malignant potential. Its definite diagnosis depends on pathological examination. Up to now, early surgery and combined therapy are effective treatments for prostatic stromal tumor of uncertain malignant potential.

[Surgical treatment of the complex bicondylar tibial plateau fracture using a midline longitudinal incision].

Objective: To explore the outcomes after treatment of the complex bicondylar tibial plateau fracture through a midline longitudinal approach. Methods: A review of fifteen patients with an average age of (51.3±12.3) years old (range17-65yers;7 males, 8 females) surgically treated from October 2013 to Febuary 2018 were included. Sahatker Ⅴ in 10, Sahatker Ⅵ in 5; fractures of medial and lateral columns in 9, fractures of three columns in 6. All the patients were adopt a midline longitudinal approach combined with the posterior approach and bone grafting were conducted. Results: All cases were followed-up for (14.4±3.8) month, with an average of 12-24 month. All patients gained bone union during 12-16 weeks after operation, with an average of (15.2±1.3) weeks. There were significant differeces in both tibial plateau angle and posterior slope angle on radiography between preoperation and postoperation (P<0.05), there were no significant differeces in either tibial plateau angle or posterior slope angle on radiography between immediate postoperation and 12 months postoperation (P>0.05). At final follow-up,both the Lachman test and the Pivot-shift test were negative. All patients had complete knee extension, knee flexion angle 100°-135°, with an average of 117.7°±11.3°. The HSS (the Hospital for Special Surgery) score were 66-98, with an average of 85.1±9.3, six cases were excellent and seven cases were good, two cases was fair, the excellent and good rate was 86.7%. The Rasmussen radiological evaluationre were 9-18, with an average of 15.1±2.5, three cases were excellent and eleven cases were good, one cases was fair, the excellent and good rate was 93.3%. 1 patient had fat liquefactionof in antero incision, and got good outcomes after debridement dressing. Conclusion: The treatment of the complex bicondylar tibial plateau fracture through a midline longitudinal approach combined with the posterior approach can result in good exposure and satisfying knee function in short-term.

Real-Time Trafficking of Agrobacterium Virulence Protein VirE2 Inside Host Cells.

A. tumefaciens delivers T-DNA and virulence proteins, including VirE2, into host plant cells, where T-DNA is proposed to be protected by VirE2 molecules as a nucleoprotein complex (T-complex) and trafficked into the nucleus. VirE2 is a protein that can self-aggregate and contains targeting sequences so that it can efficiently move from outside of a cell to the nucleus. We adopted a split-GFP approach and generated a VirE2-GFP fusion which retains the self-aggregating property and the targeting sequences. The fusion protein is fully functional and can move inside cells in real time in a readily detectable format: fluorescent and unique filamentous aggregates. Upon delivery mediated by the bacterial type IV secretion system (T4SS), VirE2-GFP is internalized into the plant cells via clathrin adaptor complex AP2-mediated endocytosis. Subsequently, VirE2-GFP binds to membrane structures such as the endoplasmic reticulum (ER) and is trafficked within the cell. This enables us to observe the highly dynamic activities of the cell. If a compound, a gene, or a condition affects the cell, the cellular dynamics shown by the VirE2-GFP will be affected and thus readily observed by confocal microscopy. This represents an excellent model to study the delivery and trafficking of an exogenously produced and delivered protein inside a cell in a natural setting in real time. The model may be used to explore the theoretical and applied aspects of natural protein delivery and targeting.

The novel Jab1 inhibitor CSN5i-3 suppresses cell proliferation and induces apoptosis in human breast cancer cells.

Jab1 is a vital subunit of the CSN family and is reported to be overexpressed in numerous cancer types. Due to the importance of Jab1/CSN5 in cancer cell proliferation and survival, Jab1 is considered a promising therapeutic target. Therefore, we evaluated the anticancer effect of the novel Jab1 inhibitor CSN5i-3 in breast cancer cells. In our study, we found that Jab1 was overexpressed in breast cancer tissues and was correlated with poor prognosis in human breast cancer patients. An MTS assay revealed that CSN5i-3 suppressed cell proliferation in the breast cancer cell lines BT474 and SKBR3. We also found that CSN5i-3 significantly induced apoptosis and G1 phase cell cycle arrest in breast cancer cells. A mechanistic investigation revealed that CSN5i-3 inhibited Jab1 expression and increased the level of the apoptosis marker cleaved PARP and the cell-cycle-related protein p27 in BT474 and SKBR3 cells. A nude mouse xenograft model also indicated that CSN5i-3 exerted a potent anticancer effect in vivo. Overall, our study suggested that the Jab1 inhibitor CSN5i-3 might be a promising agent for the treatment of breast cancer in humans and should be studied further.

Evaluating the Efficiency of REPLI-g® Single Cell Kit for Trace DNA Amplification.

ABSTRACT: Objective To evaluate the efficiency of REPLI-g® Single Cell Kit for sample DNA amplification, and explore its application value in forensic trace DNA amplification. Methods Three DNA extraction kits were selected to extract DNA from peripheral blood of 10 unrelated individuals. The DNA yield and purity of the three DNA extraction kits were compared. According to the results of comparison, one DNA sample was selected to concentrate and dilute, then used as the initial sample of whole genome amplification （WGA）. REPLI-g® Single Cell Kit was used to amplify the initial sample at the whole genome level. The amplification yield and amplification times were calculated, and the distribution of DNA fragments was detected by agarose gel electrophoresis. Goldeneye® DNA ID System 20A Kit was used to perform the STR typing of the initial sample and DNA samples amplified at the whole genome level to evaluate the performance of REPLI-g® Single Cell Kit in trace DNA amplication in terms of purity and yield as well as the success rate of STR typing. Results After comparison, one DNA sample was selected from QIAsymphony® DNA Investigator® Kit extracts to concentrate and dilute as the initial sample of WGA. After amplifying the whole genome of a series of initial samples by REPLI-g® Single Cell Kit, the lowest average of amplification yield reached 8.77×103 ng, while the average of the corresponding amplification times reached 1.40×106. DNA fragments were large and concentrated. The STR typing success rate of WGA samples became lower with the decrease of initial samples used, but when the amount of samples was lower than 0.5 ng, the STR typing success rate of samples after DNA WGA was higher than that of samples without DNA WGA. Conclusion REPLI-g® Single Cell Kit can increase the yield of template DNA. Especially for trace DNA, the STR typing success rate can be improved to a certain extent.

Th17, synchronically increased with Tregs and Bregs , promoted by tumour cells via cell-contact in primary hepatic carcinoma.

Documented reports about T helper type 17 (Th17) cells have revealed that Th17 plays a critical role in inflammation and autoimmunity diseases. However, the role of Th17 in cancer remains contradictory. The interplay between Th17 and tumour cells in the tumour microenvironment of primary hepatic carcinoma (PHC) needs to be explored further and the relationship between Th17, regulatory T cells (Tregs ) and regulatory B cells (Bregs ) has not been defined completely. In this study, numerous experiments were undertaken to elucidate the interaction of Th17 and Treg /Breg cells involved in PHC. Our work demonstrated that an increased Th17 was detected in the peripheral circulation and in tumour tissues in PHC patients. In addition, increases in peripheral blood Th17 corresponded with tumour-node-metastasis (TNM) stage progression. Also, further studies indicated that Th17 cells were promoted by tumour cells in the PHC tumour microenvironment through both contact-dependent and -independent mechanisms, but cell-contact played the major important role in promoting the production and proliferation of Th17. When isolated CD4+ CD25+ CD127low Tregs and CD4+ CD25- CD127+ non-Tregs were cultured with autologous tumour cells, it implied that the phenotype of Th17 and Tregs was modified by tumour cells in the tumour microenvironment. As well as this, Th17 cells were also found to correlate positively with CD4+ forkhead box protein 3+ Tregs and CD19+ CD5+ CD1dhi Bregs in PHC. Notably, Th17 increased synchronically with Tregs and Bregs in PHC. These findings may provide new clues to reveal the mechanisms of immune escape in PHC.

Optimization of the medium for the growth of Nitrobacter winogradskyi by statistical method.

Although Nitrobacter winogradskyi is an important chemoorganotrophic organism for the study of nitrite-oxidizing bacteria physiology as well as nitrification, until now, the mixotrophic medium for this organism growth has not been optimized, comprehensively. In this study, we aimed to improve the growth medium of N. winogradskyi using the one-factor-at-a-time (NaNO2 , glycerol, pH) method. In addition, a further experimental design was carried out based on central composite design with response surface methodology. Different combinations of the three cultural parameters were fitted by multiple regression analysis to calculate the predicted response. Our results suggest that optimal culture condition for the growth of N. winogradskyi was a modified DSMZ 756a medium containing NaNO2 (5·74 g l-1 ) and glycerol (37·88 mmol l-1 ), pH 7·83, a temperature of 28°C and agitation at 120 rev min-1 . The results from a validation experiment (bacterial growth: OD600 1·0293) were close to the value predicted by the quadratic model (OD600 1·0994). In addition, we uncovered the potential mechanism at the cellular and ultrastructural levels. The results indicated that glycerol in the media enhanced the rate of cell division and cell growth by increasing the accumulation of polyphosphates and phosphorus, and high concentrations of NaNO2 provided sufficient energy for growth and contributed to the generation of carboxysomes in cells for CO2 fixation. SIGNIFICANCE AND IMPACT OF THE STUDY: Due to the extremely slow growth rate and the low growth yield of ammonia-oxidizing bacteria and NOB (nitrite-oxidizing bacteria), nitrification is still the rate-limiting step of nitrogen cycle in the current research. Nitrobacter winogradskyi, an important NOB, participates in the second step of nitrification in water and soil. This study reported an optimized culture condition for N. winogradskyi, which increased the growth yield by 5·06 times than that in the basal medium and uncovered the potential mechanism. We expect our study will contribute to the research on water and soil nitrogen cycle. In addition, the optimized culture conditions have the potential to be suitable for the chemoorganotrophic growth of other nitrifiers.

Regulatory functions of trehalose-6-phosphate synthase in the chitin biosynthesis pathway in Tribolium castaneum (Coleoptera: Tenebrionidae) revealed by RNA interference.

RNA interference (RNAi) is a very effective technique for studying gene function and may be an efficient method for controlling pests. Trehalose-6-phosphate synthase (TPS), which plays a key role in the synthesis of trehalose and insect development, was cloned in Tribolium castaneum (Herbst) (TcTPS) and the putative functions were studied using RNAi via the injection of double-stranded RNA (dsRNA) corresponding to conserved TPS and trehalose-6-phosphate phosphatase domains. Expression analyses show that TcTPS is expressed higher in the fat body, while quantitative real-time polymerase chain reaction results show that the expression of four trehalase isoforms was significantly suppressed by dsTPS injection. Additionally, the expression of six chitin synthesis-related genes, such as hexokinase 2 and glutamine-fructose-6-phosphate aminotransferase, was suppressed at 48 and 72 h post-dsTPS-1 and dsTPS-2 RNA injection, which were two dsTPS fragments that had been designed for two different locations in TcTPS open reading frame, and that trehalose content and trehalase 1 activity decreased significantly at 72 h post-dsRNA injection. Furthermore, T. castaneum injected with dsTPS-1 and dsTPS-2 RNA displayed significantly lower levels of chitin and could not complete the molting process from larvae to pupae, revealing abnormal molting phenotypes. These results demonstrate that silencing TPS gene leads to molting deformities and high mortality rates via regulation of gene expression in the chitin biosynthetic pathway, and may be a promising approach for pest control in the future.