AAV9-based PMM2 gene replacement augments PMM2 expression and improves glycosylation in primary fibroblasts of patients with phosphomannomutase 2 deficiency (PMM2-CDG).

Inherited deficiency of phosphomannomutase 2 (PMM2) (aka PMM2-CDG) is the most common congenital disorders of glycosylation (CDG) and has no cure. With debilitating morbidity and significant mortality, it is imperative to explore novel, safe, and effective therapies for the disease. Our Proof-of-Concept study showed that AAV9-PMM2 infection of patient fibroblasts augmented PMM2 expression and improved glycosylation. Thus, AAV9-PMM2 gene replacement is a promising therapeutic strategy for PMM2-CDG patients.

[A multicenter cross-sectional study of quality of life and nonsurgical treatment in patients with knee osteoarthritis].

Objective: To explore the influencing factors of health-related quality of life (HRQoL) in patients with knee osteoarthritis, and to analyze the non-surgical treatment of knee osteoarthritis. Methods: Demographic variables, treatment modalities, imaging data, and 12-item short form health survey (SF-12) scores of patients with knee osteoarthritis in orthopedic outpatient departments of five hospitals in Beijing from December 2017 to November 2018 were collected to analyze influencing factors of HRQoL and non-surgical treatment. Results: A total of 2 034 patients were included. There were 530 males (26.1%) and 1 504 females (73.9%), with a mean age of (59.17±10.22) years. In terms of physical quality of life, female patients with knee osteoarthritis had lower physical components summary (PCS) compared with male patients (ß=-0.521, P=0.036); patients aged ≥64 years had lower PCS than those aged<55 years (ß=-0.636, P=0.026). Patients with an education of more than 12 years had higher PCS than those with less than 10 years (ß=1.063, P<0.001). Compared to patients with mild clinical symptoms, the PCS of patients with moderate clinical symptoms was lower (ß=-0.860, P=0.002), while the PCS of those with severe clinical symptoms was much lower (ß=-1.126, P<0.001). Patients treated with combination therapy had higher PCS than untreated patients (ß=0.731, P=0.005). In terms of mental quality of life, compared to patients engaged in sedentary work, the mental components summary (MCS) of patients engaged in mild manual labor jobs was lower (ß=-0.712, P=0.015); Compared to patients with a Charson comorbidity index of 0, patients with a Charlson comorbidity index ≥ 2 had lower MCS (ß=-1.183, P=0.007). In the past 12 months, 648 (31.9%), 143 (7.0%), 406 (20.0%), 680 (33.4%), 343 (16.9%), 681 (33.5%), 170 (8.4%) patients had used non-steroid anti-inflammatory drugs (NSAIDs), acetaminophen, glucosamine/chondroitin formulations, physical therapy, articular cavity puncture injection, traditional Chinese medicine treatment and exercise therapy, respectively. Total of 451 patients (22.2%) received monotherapy and 889 patients (43.7%) received combination therapy. Conclusions: The major non-surgical treatment methods for patients with knee osteoarthritis in Beijing are NSAIDs, physiotherapy and traditional Chinese medicine. Combination therapy is used more frequently than monotherapy. Physical quality of life is related to gender, age, education, severity of symptoms and treatment, while mental quality of life is related to occupational labor and comorbidities.

Analysis of Approaches in the Microsurgical Treatment of 102 Cases of Petroclival Meningioma in a Single Center.

Objectives: We identified the optimal approaches for treating the diverse tumor subtypes of petroclival meningioma (PM) by analyzing the clinical benefits of various surgical approaches adopted for each subtype. Methods: Tumors in 102 PM patients from a single center who underwent surgical treatment were classified as upper clivus (UC), cavernous sinus (CS), tentorium (TE), or petrous apex (PA) types based on the attachment site of the tumor base and the displacement of the trigeminal nerve. The therapeutic effects of different surgical approaches among the subtypes were evaluated according to the patient outcomes. Results: The subtemporal (33.33%), retrosigmoid (16.67%), and Kawase approaches (50%) were used for the UC type. Simpson I/II resection was achieved in 46.66% of patients with the Kawase approach. Significant differences were found between the other two approaches (P = 0.044) and in the follow-up Karnofsky performance scale (KPS) scores (P = 0.008). The subtemporal (60%) and Kawase approaches (40%) were used for the CS type; neither approach achieved Simpson I/II resection. The retrosigmoid (25.81%) and Kawase approaches (74.19%) were used for the TE type. The Simpson I/II resection rates of the two approaches were 55.55 and 86.95%, respectively, and a significant difference was observed between them (P = 0.039). The retrosigmoid (43.75%) and Kawase approaches (56.25%) were used for the PA type. The Simpson I/II resection rates of the two approaches were 31.25 and 50%, respectively. The resection degrees of the two approaches and the KPS scores at follow-up were significantly different (P = 0.034). Conclusion: The individual microsurgical approaches adopted for the various PM tumor subtypes can provide maximal safe resection and good KPS scores. The Kawase approach is more suitable for PM, especially for UC- and PA-type PM tumors.

Clinical and biological impact of LINC02544 expression in breast cancer after neoadjuvant chemotherapy.

OBJECTIVE: Analysis of breast cancer and cancer tissue after neoadjuvant chemotherapy (nCT) may be helpful to find new biomarkers. It is known that long non-coding RNAs (lncRNAs) are involved in the carcinogenic pathway and drug resistance of breast cancer. Our aim was to determine the role of LINC02544 in the behavior and outcome of post-nCT breast cancer. PATIENTS AND METHODS: The expression level of LINC02544 in breast cancer and its effect on the survival time of patients were predicted by lnCAR database. In vitro, EdU, Wound-healing and transwell assays were used to detect the effects of LINC02544 on the proliferation, migration and invasion of MCF-7 cells, and the related regulatory networks were analyzed by the database. Quantitative Reverse Transcriptase-Polymerase Chain Reaction (QRT-PCR) was used to detect the expression of LINC02544 in 147 cases of nCT before and after treatment, and the relationship between the expression of LINC02544 and survival time and clinicopathological features was analyzed. RESULTS: LINC02544 was highly expressed in breast cancer and led to poor prognosis. Overexpression of LINC02544 promoted proliferation, invasion and migration of breast cancer cells. Compared with the residual tumor after nCT with low expression of LINC02544, the high expression of LINC02544 in the residual tumor after nCT was significantly correlated with overall survival and disease-free survival. CONCLUSIONS: In this study, it is suggested that LINC02544 has a potential application prospect as a biomarker and therapeutic target for breast cancer patients and neoadjuvant therapy patients.

Metformin Ameliorates Lipopolysaccharide-Induced Depressive-Like Behaviors and Abnormal Glutamatergic Transmission.

Metformin, a first-line drug for type 2 diabetes mellitus (T2DM), has been found to reduce depressive symptoms in patients with comorbid depression and other diseases. However, it is largely unclear how metformin ameliorates depressive-like behaviors. Here, we used lipopolysaccharide (LPS) to induce depressive-like behaviors in mice and found that LPS-treated mice exhibited increased immobility in the forced swimming test (FST) and tail suspension test (TST), as well as increased glutamatergic transmission. Furthermore, metformin administration in the LPS-treated mice ameliorated depressive-like behaviors and elevated glutamatergic transmission. Our results suggest that metformin has antidepressant effects and can correct abnormal glutamatergic transmission, providing an insight into the underlying mechanism by which metformin acts against depression.

Whole-genome resequencing of Dulong Chicken reveal signatures of selection.

1. Dulong Chickens (DLCs) live at high altitude (~3000 m) and humidity (~90%), are endemic to the Yunnan province, and have gradually developed unique physiological characteristics, but their genetic basis is still unclear. Using the fixation index (FST ) approach, based on whole-genome resequencing, DLCs were analysed to uncover the genomic architecture of the population and candidate genes involved in selection during domestication. 2. A total of 469 candidate genes were obtained to be putatively under selection in DLCs. Further investigations revealed the genic footprint for local adaptation (high-altitude and high-humidity) as the genic signatures that are involved in economic traits (related to egg production). 3. Candidate genes were identified that may be associated with disease resistance, aggressiveness, small body size and positive selection of vision in DLCs. 4. These data revealed loci of selective signals that operate during selection for production at high altitude and humidity.

[Application value of diffusion-weighted imaging signal pattern in the qualitative diagnosis of renal lesions with T(2)WI hypointensity].

Objective: To analyze the diagnostic value of diffusion-weighted imaging (DWI) signal pattern in renal lesions with T(2)WI hypointensity. Methods: Retrospective analysis was performed on DWI imaging datasets of 135 renal lesions with hypointentsity on T(2)WI confirmed by surgery, biopsy, or follow-up in the First Medical Center of Chinese PLA General Hospital from February 2016 to February 2017.One hundred and thirty-five renal lesions,43 benign lesions(age from 28 to 70 years,mean age was 43.5 years, male 18 lesions and female 25 lesions) and 92 malignant lesions (age from 17 to 86 years, mean age was 54.1 years, male 62 lesions and female 30 lesions). DWI signal pattern was classified into six categories: homogeneously high signal, homogeneously low signal, heterogeneously high signal, high halo signal, high halo and nodular signal, and high nodular signal. The agreement between two observers were tested using kappa statistic. The statistical difference between DWI signal characteristics in benign and malignant lesions was analyzed with Chi-Square test. Diagnostic efficacy in differentiation of benign and malignant renal lesions using DWI signal pattern were calculated. Results: One hundred and thirty-five lesions were detected in 135 cases with T(2)WI hypointensity. There were 43 benign lesions and 92 malignant lesions. The agreement between two observers was very good (kappa value=0.878 6). In renal T(2)WI hypointensity lesions, the proportion of DWI homogeneous high signal, homogeneous low signal, heterogeneous high signal, high halo signal, high halo and nodular signal, high nodular signal was 4.7% (2/43), 25.6% (11/43), 30.2% (13/43), 18.6% (8/43), 11.6% (5/43), 9.3% (4/43), respectively. The proportion of malignant lesions was 10.9% (10/92), 0 (0/92), 17.4% (16/92), 13.0% (12/92), 56.5% (52/92) and 2.2% (2/92), respectively. The difference of high halo and nodules signal and homogeneous low signal was statistically significant (all P<0.01). The sensitivity, specificity, positive prediction value (PPV) and negative prediction value (NPV) of high halo and nodular signal for malignancy were 56.5% (52/92), 88.4%(38/43), 91.2% (52/57) and 48.7%(38/78), respectively and homogeneous low signal for benign lesions were 25.6% (11/43), 100.0% (92/92), 100.0% (11/11) and 74.2% (92/124), respectively. Conclusions: DWI signal features may facilitated the accurate diagnosis of renal lesions with T(2)WI hypointensity. Malignant lesions exhibit a higher propensity with high halo and nodular signal on DWI while benign lesions with homogeneous low signal.

MiR-99a suppresses cell migration and invasion by regulating IGF1R in gastric cancer.

OBJECTIVE: Gastric cancer is a common kind of gastrointestinal malignancies. Increasing evidence indicates dysregulation of microRNA-99a (miR-99a) in gastric cancer, and has been extensively investigated in terms of cancer formation, progression, diagnosis, therapy, and prognosis. The purpose of this study is to explore how miR-99a worked in gastric cancer on migration and invasion. PATIENTS AND METHODS: The mRNA and protein levels of miR-99a and insulin-like growth factor 1 receptor (IGF1R) in gastric cancer were measured by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot. Transwell assay was employed to analyze the migratory and invasive capacities. The Dual-Luciferase reporter assay was performed to confirm miR-99a mediated the expression of IGF1R by directly targeting its mRNA 3'-untranslated regions (3'-UTR) in gastric cancer cells. RESULTS: MiR-99a was discovered to be significantly downregulated while IGF1R was upregulated in gastric cancer tissues and cell lines. The expression of miR-99a had a negative correlation with the IGF1R expression in gastric cancer tissues. Moreover, miR-99a was low expressed in gastric cancer cells HGC-27 and MGC-803 compared to the normal cell line. MiR-99a suppressed the migration and invasion through directly binding to the 3'-UTR of IGF1R mRNA in HGC-27 cells. In addition, IGF1R could reverse partial roles of miR-99a on migration and invasion in gastric cancer. CONCLUSIONS: MiR-99a inhibited the migratory and invasive abilities by regulating the expression of IGF1R. MiR-99a was downregulated while IGF1R was upregulated in gastric cancer cell lines. The newly identified miR-99a/IGF1R axis provides novel insight into the pathogenesis of gastric cancer.

[Microbiology analysis of periprothetic joint infection post total hip and knee arthroplasty of 9 centers in Beijing between 2014 and 2016].

Objective: To investigate the microbiological test, antibiotic sensitivity and surgical treatment of periprosthetic joint infection(PJI) cases in post total hip arthroplasty (THA) and total knee arthroplasty (TKA) patients. Methods: A retrospective cross-sectional survey was conducted on 318 patients who underwent THA or TKA in 9 clinical centers in Beijing from January 2014 to December 2016.The data of microbiology, antibiotic sensitivity and surgical treatment were collected.The average age of patients was (62.3±13.1) years old (range: 21-86 years old), including 145 males and 173 females.The body mass index was (25.6±3.8) kg/m (2) (range: 15.6-38.1 kg/m(2)). Results: In total, 318 patients had microorganisms detected by periprosthetic tissue culture or synovial fluid culture, 209 cases (65.7%) had Gram-positive bacteria, 29 cases (9.1%) had Gram-negative bacteria, 10 cases (3.1%) had fungi, 3 cases (0.9%) had non-tuberculous mycobacteria, 72 cases (22.6%) were negative, 69 cases (21.7%) had methicillin-resistant bacteria. The antibiotic sensitivity results showed that the overall resistance rate of penicillin, cefuroxime, amoxicillin+clavulanic acid was 79.9%, 69.9%, and 68.1%, respectively; meropenem, vancomycin, and linezolid resistance rate was 0. For the treatment methods of hip and knee PJI, two-stage revision surgery acounted for 72.9% (108/148) and 64.1% (109/170), respectively. One-stage revision surgery accounted for 21.6% (32/148) and 7.6% (13/170), and open debridement surgery accounted for 4.7%(7/148) and 26.4% (45/170). Conclusions: Gram-positive bacteria was still the main pathogen of PJI.The methicillin-resistant bacteria and rare bacteria should be payed attention to. The Majority of hip and knee PJI cases were treated by two-stage revision surgery.

[Application of multimode MRI in prediction of nuclear grade of clear cell renal cell carcinoma].

Objective: To investigate the predictive value of multimode MRI features for nuclear grade of clear cell renal cell carcinoma (ccRCC). Methods: From January 2016 to October 2017, 381 patients (387 tumors) with ccRCC proven by pathology in Chinese PLA General Hospital First Medical Center were enrolled (male 293, female 88, age 24-87 years old). The clinical and imaging data of these patients were retrospectively analyzed, including clinical information (gender, age, BMI, smoke, hypertension) and preoperative renal MRI. Pre-and post-contrast MRI features were subjectively scored. The largest diameter of each lesion was measured. Two-sample t-test,Chi-squared test and binuary Logistic regression analysis were used to evaluate the predictive efficacy of clinical and MRI data. Results: According to WHO/ISUP nuclear grade system,all ccRCCs (n=387) were divided into low grade (n=322) and high grade group (n=65). Between two groups, there were significant differences in age and diameter((54±12) vs (59±10) years old, P=0.001; (4.1±2.2) vs (6.2±3.0) cm, P<0.01). In MRI scores,there were significant differences in scores of pseudocapsule, shape and margin,hemorrhage,enhancement degree,cystic-solid,intratumoral vessel,peritumoral vessel, renal sinus invasion, vein thrombosis, lymphadenopathy, necrosis, perinephric invasion and metastasis, DWI signal intensity between high grade group and low grade group (all P<0.01). Binuary Logistic regression analysis showed that shape and margin, enhancement degree and DWI signal intensity were independent predictors for high grade ccRCC (OR=0.181, 95%CI 0.049-0.666; OR=0.393, 95%CI 0.182-0.846; OR=0.336, 95%CI 0.155-0.728). A nomogram model for predicting the risk of high grade ccRCC was constructed. Conclusions: Multimode MRI features can differentiate low grade and high grade ccRCC. The nomogram developed in this study might aid urologist in the pre-operative prediction of nuclear grade of ccRCC,which might contribute to developing treatment strategy.

Research on miR-126 in glioma targeted regulation of PTEN/PI3K/Akt and MDM2-p53 pathways.

OBJECTIVE: To study the correlations of micro ribonucleic acid (miR)-126 expression with pathogenesis and prognosis of glioma, and to screen potential biological targets for the diagnosis, treatment and prognosis of glioma. PATIENTS AND METHODS: miR-126 expression in cancer tissues, normal brain tissues, U87MG cells and normal astrocytes in glioma patients was quantitatively analyzed via quantitative reverse transcription-polymerase chain reaction (qRT-PCR). U87MG cells were transfected with miR-126 mimics or miR-126 inhibitor, followed by verification via qRT-PCR. The cell proliferation, apoptosis, migration and invasion after transfection were analyzed using methyl thiazolyl tetrazolium (MTT) assay, Annexin V/propidium iodide (PI) assay, wound healing assay and transwell assay, respectively. The expression levels of proteins related to phosphatase and tensin homolog deleted on chromosome ten/phosphatidylinositol 3-kinase/protein kinase B (PTEN/PI3K/Akt) pathway and double mouse minute 2 homolog (MDM2)-p53 pathway were detected via Western blotting. Moreover, the prognostic analysis was performed using the Kaplan-Meier method and log-rank test. RESULTS: Results of qRT-PCR showed that the miR-126 expression in highly malignant glioma tissues and U87MG cells were significantly lower than those in normal brain cells, and its expression level was significantly higher or lower than that in negative control group after transfection with miR-126 mimics or inhibitor. Analyses of cell proliferation, apoptosis, migration and invasion revealed that the up-regulation of miR-126 could remarkably inhibit the in-vitro proliferation, migration and invasion and promote apoptosis of glioma cells, and vice versa. Results of Western blotting manifested that after overexpression of miR-126, PI3K, p-Akt and MDM2 protein levels in U87MG cells were significantly decreased compared with those in control group, but PTEN and p53 protein expressions were significantly increased, and vice versa. Besides, according to prognostic analysis, the prognosis of patients with a low miR-126 level was poorer. CONCLUSIONS: The miR-126 expression is abnormally low in glioma cells, and miR-126 inhibits the course of glioma through targeted regulation of PTEN/PI3K/Akt and MDM2-p53 pathways, which, therefore, can be used as a new potential biomarker for the diagnosis, treatment and prognosis of glioma.

[Magnetic resonance imaging findings and differential diagnosis of renal epithelioid angiomyolipoma comparing with renal no-epithelioid angiomyolipoma].

Objective: To investigate whether MRI findings can differentiate renal epithelioid angiomyolipoma (EAML) from renal no-epithelioid (typical) angiomyolipoma. Methods: A total of 44 patients were collected from General Hospital of PLA.These cases were obtained from January 2009 to June 2015.To retrospectively analyze these mainly MRI findings among 12 cases of EAML (age from 27 to 61 years, male 2 cases, female 10 cases, mean age was 46.7 years); 32 cases of renal no-epithelioid AML (age from 34 to 70 years old, male 9 cases, female 23 cases, mean age was 53.4 years old) as case control study. MRI findings included gender, T(2)WI, the signal on gross fat, pseudocapsle, necrosis or cystic degeneration, DWI, hemorrhage and the peak in three phases of dynamic enhancement.All data were analyzed statistically using SPSS version 19.0 (IBM, Armonk, NY, USA). χ(2) test and a single order chart were used to analyze the enumeration data. Results: Comparing with renal no-epithelioid angiomyolipoma, minimal fat, necrosis or cystic degeneration and hemorrhage were statistical significance. P values were 0.002, 0.007, 0.025, respectively.Gender, solid components of tumors on T(2)WI, the signal of DWI, pesudocapsule and the peak of enhancement had no statistical significance. P values were 0.863, 0.053, 0.479, 0.460, respectively. Conclusion: Comparing with renal no-epithelioid AML, necrosis or cystic degeneration and hemorrhage with minimal fat are characteristic MRI findings of EAML.

The challenge of diagnosing seborrheic keratosis by reflectance confocal microscopy.

BACKGROUND: Seborrheic keratosis (SK) is one of the most common skin tumors seen by dermatologists. It should be differentiated with many diseases, especially skin tumors. Reflectance confocal microscopy (RCM) has been applied for evaluation of SK. There are a few studies that describe the RCM of SK. The aim of the study was to find the challenge of diagnosing seborrheic keratosis by reflectance confocal microscopy. METHODS: A total of 390 patients with a clinical suspicious diagnosis of seborrheic keratosis were enrolled in this study, and lesions from each patient were imaged with RCM. Thirty-seven of these patients performed a biopsy in order to be given a histological diagnosis. We retrospectively analyzed the outcomes of RCM diagnosis and histological diagnosis, and then found the RCM characteristics of biopsy-proven lesions. RESULTS: According to RCM images, 258 of 390 (66.2%) patients were diagnosed with SK, 97 of 390 (24.9%) patients could not be diagnosed by the dermatologist according to RCM. Of all 37 biopsied lesions, 23 were SK, 6 were actinic keratosis, 2 were basal cell carcinoma, and 2 were squamous cell carcinoma. CONCLUSION: It is challenge to diagnose seborrheic keratosis by reflectance confocal microscopy. It may due to the variable clinical and RCM appearances of SK, and limited depth of RCM.

Early immune anergy towards recall antigens and mitogens in patients at onset of septic shock.

The pathology of sepsis is typically characterized by an infection and excessive initial inflammation including a cytokine storm, followed by a state of immune suppression or paralysis. This classical view of a two peak kinetic immune response is currently controversially discussed. This study was a sub-study of the randomized clinical Trial SISPCT registered with www.clinicaltrials.gov (NCT00832039, Registration date: 29/01/2009). Blood samples from 76 patients with severe sepsis and septic shock were incubated for 48 h at 37 °C in vitro with bacterial or fungal recall-antigens or specific mitogen antigens within 24 hours of sepsis onset. Recall-antigen stimulation led to a severe dampening of normal cytokine release. This immunologic anergy was similarly observed after mitogen stimulation. Moreover, patients under hydrocortisone therapy or with lowered arterial oxygen tension had further reductions in cytokine levels upon B- and T-cell mitogen stimulation. This investigation reveals an early onset of immunoparalysis during sepsis. This immune incompetence in mounting an adequate response to further infections includes previously sensitized pathogens, as seen with recall-antigens. Also, the immune-suppressive role of hydrocortisone and low PaO2 is highlighted. Aside from early broad-spectrum antimicrobial therapy, our findings reinforce the need for maximal immunological support and protection against further infections at the onset of sepsis.