[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.

[Effect of posterior tibial slope on the short-term outcome in mobile-bearing unicompartmental knee arthroplasty].

OBJECTIVE: To evaluate the relationship between postoperative knee function and the sagittal position of tibial component in unicompartmental knee arthroplasty (UKA). METHODS: We retrospectively enrolled the patients who underwent UKA from January 2016 to May 2020. They were assigned into 2 groups according to postoperative posterior tibial slope (PTS): the normal PTS group (PTS≥3° and PTS < 8°) and the abnormal PTS group (PTS < 3° or ≥8°). The patients were followed up for at least 12 months. The postoperative Knee Society Clinical Score (KSS-C), Knee Society Functional Score (KSS-F) and knee range of motion (ROM) were compared between the two groups. RESULTS: A total of 72 patients (82 knees) were included with 51 patients (58 knees) in PTS normal group and 21 patients (24 knees) in PTS abnormal group. All the patients were followed up with median of 23.6 months. There was no significant difference in the general data [gender, age, body mass index (BMI)], pre-operative knee range of motion, preoperative KSS-C score and KSS-F score (P > 0.01). The KSS-C score, KSS-F score, and knee range of motion significantly improved after surgery (P < 0.01) for all the patients. The postoperative KSS-C score in normal PTS group (88.76±2.79) was significantly higher than the KSS-C score in abnormal PTS group (84.42±3.35, P < 0.01), but no significant difference between the 2 groups was observed in postoperative KSS-F score and knee range of motion (P > 0.01). In addition, there was no correlation between the change of PTS and postoperative KSS-C score (r=-0.034, 95%CI: -0.247 to 0.186, P = 0.759), KSS-F score (r = -0.014, 95%CI: -0.238 to 0.198, P = 0.901) and knee range of motion (r= 0.045, 95%CI: -0.214 to 0.302, P = 0.686). CONCLUSION: The posterior tibial slope between 3° and < 8° can be recommended to improve knee joint function in mobile UKA, and excessive or insufficient PTS should be avoided.

The key aroma compounds and sensory characteristics of commercial Cheddar cheeses.

To study the key aroma components and flavor profile differences of Cheddar cheese with different maturity and from different countries, the flavor components of 25 imported commercial Cheddar cheese samples in the China market were determined by gas chromatography-mass spectrometry. The quality and quantity of 40 flavor compounds were analyzed by gas chromatography-olfactometry among 71 aroma compounds determined by gas chromatography-mass spectrometry. Combined with odor activity value calculation, principal component analysis (PCA) was conducted to analyze the relationship among 26 flavor compounds with odor activity values >1 and the maturity of Cheddar cheese. The PCA results showed significant differences between the group of mild Cheddar cheese and the groups of medium Cheddar cheese and mature Cheddar cheese, and no significant differences were observed between medium Cheddar cheese and mature Cheddar cheese. According to the results of PCA and consumers' preference test, representative Cheddar cheese samples with different ripening times were selected for the flavor profile analysis. Partial least squares regression analysis was conducted to obtain the relationship between sensory properties and flavor compounds of different Cheddar cheeses. Based on partial least squares regression analysis, 1-octen-3-one, hexanal, acetic acid, 3-methylindole, and acetoin were positively correlated with milky, sour, and yogurt of mild Cheddar cheese. Dimethyl trisulfide, phenylacetaldehyde, ethyl caproate, octanoic acid, and furaneol and other compounds were positively correlated with fruity, caramel, rancid, and nutty notes of the medium and mature Cheddar cheeses.

Key aroma compounds identified in Cheddar cheese with different ripening times by aroma extract dilution analysis, odor activity value, aroma recombination, and omission.

To determine the odor-active compounds in Cheddar cheeses with different ripening times (6, 10, and 14 mo), 39 potent odorants of Cheddar cheeses were identified with a flavor dilution factor range between 1 and 512 by aroma extract dilution analysis. To further determine their contribution to the overall aroma profile of Cheddar cheeses, odor activity values of 38 odorants with flavor dilution factors ≥1 were calculated. A Cheddar cheese matrix was developed to determine the concentrations and the odor thresholds of these key aroma compounds. The result of the aroma recombinant experiment prepared by mixing the key aroma compounds in the concentrations in which they occurred in Cheddar cheeses showed that the overall aroma profile of the recombinant sample was very similar to that of Cheddar cheese. The main different compounds in Cheddar cheese with different ripening time were acetic acid, butanoic acid, dimethyl trisulfide, methional, hexanal, (E)-2-nonenal, acetoin, 1-octen-3-one, δ-dodecalactone, furaneol, hexanoic acid, heptanal, and ethyl caproate. This study could provide important information for researching and developing Cheddar cheese-related products.

SCARA5 is a Novel Biomarker in Colorectal Cancer by Comprehensive Analysis.

BACKGROUND: SCARA5 has been demonstrated to be a tumor suppressor gene, with its expression downregulated in many cancer types. However, only few studies have investigated its role in colorectal cancer (CRC). The current study evaluated SCARA5 expression levels in CRC and its potential value as a diagnostic biomarker for CRC. METHODS: Data were downloaded from the TCGA, GEO, and Oncomine databases to evaluate SCARA5 mRNA expression levels in CRC. The prognosis value of SCARA5 was assessed using the online tool Cutoff Finder via the Kaplan-Meier plotter (n = 484). Immunohistochemistry was performed to analyze and compare the SCARA5 protein expression levels in CRC and normal tissues from 67 CRC clinical specimens. Relevant CRC CNV data were downloaded from TCGA and cBioPortal for Cancer Genomics databases to assess the associated genetic alterations. GSEA was used to explore the underlying molecular mechanisms of SCARA5. The correlation between SCARA5 mRNA levels and cell cycle-associated genes was explored using GEPIA database. RESULTS: SCARA5 mRNA levels were found to be downregulated in CRC tissues compared with normal tissues. Survival analysis showed that low SCARA5 expression was associated with poor prognosis. These results were validated in clinical specimens, wherein the SCARA5 protein levels were significantly downregulated in CRC tissues compared with paracarcinoma tissues. Deep deletion was the most common genetic alteration and was consistent with the downregulated SCARA5 expression in CRC tissues. GSEA indicated that the gene sets of CELL CYCLE, G2M CHECKPOINT, and E2F TARGETS were negatively related to SCARA5 mRNA expression. GEPIA indicated that the mRNA expression of some cell cycle-associated genes was negatively correlated with that of SCARA5 in CRC. CONCLUSIONS: Thus, SCARA5 may act as a human cancer suppressor gene in CRC, and its expression level may be a reliable adjuvant parameter to diagnose CRC and predict tumor metastasis and prognosis.

Characterization of the key odorants in kurut with aroma recombination and omission studies.

Kurut is a traditional acid-coagulated cheese from the northwest region of China. Using gas chromatography olfactometry and aroma extract dilution analysis, we identified 21 potent odorants from kurut within the flavor dilution factor range of 8 to 256. We developed a kurut matrix to determine the odor thresholds of key aroma compounds in kurut. The odor activity values of these 21 potent volatile compounds revealed that 18 were present at concentrations above their odor threshold values and therefore contributed to the overall aroma of kurut. The result of aroma recombinant experiments prepared by mixing the 18 most important odorants at the concentrations in which they occurred in kurut showed that the overall aroma profile of the recombinant sample was very similar to that of kurut. Omission experiments indicated that acids, furans, esters, and ketones were the most important volatile compounds in kurut. Some aroma compounds-such as butanoic acid, homofuraneol, ethyl hexanoate, and ethyl butanoate-play the most important roles in the overall flavor profile of kurut. Some odorants with a high flavor dilution factor, such as sulfur compounds, may have little effect. The study of key aroma compounds in kurut could provide important information for researching and developing traditional Chinese cheese products.

MiR-302 a/b/c suppresses tumor angiogenesis in hepatocellular carcinoma by targeting MACC1.

OBJECTIVE: Hepatocellular carcinoma (HCC) is a hypervascularized tumor. Aberrant angiogenesis is the main cause, which results in cancer growth and progression. It has been showed that microRNA-302 cluster (miR-302) may be associated with angiogenesis. Here, we aimed to identify the role of miR-302a/b/c in the regulation of cell angiogenesis in HCC. PATIENTS AND METHODS: MRNA expression of miR-302a/b/c and MACC1 was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The protein of MACC1 was measured using Western blot. Cells proliferation, migration, and invasion abilities were investigated via Cell Counting Kit-8 (CCK-8) assay or transwell assay, respectively. Tube formatting assays were used to explore the tube formation capacity. The interaction among miR-302a/b/c was analyzed by luciferase assay. RESULTS: The expression of miR-302a/b/c was greatly reduced while MACC1 expression, whether mRNA or protein was conspicuously elevated in HCC tissues and cells. Then, functional experiment results showed miR-302a/b/c overexpression and MACC1 down-regulation inhibited the proliferation, migration, invasion ability, and tube formation capacity of HUVECs. In addition, we detected that miR-302a/b/c directly targeted MACC1 and suppressed MACC1 expression, and miR-302a/b/c could suppress tumor angiogenesis in HCC by targeting MACC1. CONCLUSIONS: MiR-302a/b/c may function as a potential suppressor of tumor angiogenesis in HCC by targeting MACC1, indicating a promising target for HCC therapy.

[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.

[Measurement of the tibial alignment after total knee replacement without the extramedullary cutting guide].

OBJECTIVE: To evaluate the accuracy and analysis of the influencing factors of the tibia prosthetic coronal alignment after total knee arthroplasty without tibia extramedullary cutting guide. METHODS: In the study, 76 patients (96 knees) who had primary total knee arthroplasty in Peking University First Hospital from February 2012 to April 2016 were selected retrospectively. All of the cases were performed by the same experienced orthopedic surgeon team. The patients were divided into 2 groups randomly: tibia cutting without guide group and tibia cutting with extramedullary guide group. Lower extremity standard anterior and posterior X-ray films of the knees were taken before and after surgery, and the anatomical tibial angle (AT), the posterior slope angle (PSA) before and after surgery, the tibia component angle (TCA) were measured in the X-ray films. We also recorded the beginning and finishing time of the operation, the operation sequence of the day respectively at the same time. The data were analyzed by correlation analysis, t test and chi square test between the two groups. RESULTS: The age, gender, body mass index, AT/PSA before the surgery and TCA/PSA after the surgery of the two groups were no significant differences (P>0.05). The rate of 90-TCA≥ 3° was 31.5% (17 knees) and 31% (12 knees) respectively, there were no significant differences (P=0.956) between the two groups. The postoperative TCA of two groups was not correlated with age, gender, body mass index (BMI), operation side (P>0.05), there was also no correlation between the postoperative TCA and the start time of the operation, the whole operation time, and the operation sequence of the day (P>0.05). The two groups were divided into subgroups according to 90-AT before the operation (0°≤90-AT<3°, 3°≤90-AT<5°, 5°≤90-AT<8°, 90-AT≥8°), and there was no difference among them. But we found there was a much higher rate of 90-TCA<3° in the group without cutting guide than the group with extramedullary guide when 90-AT≥8° before the surgery (the rates were 71.4 % and 42.9%, P<0.05). CONCLUSION: There is no significant difference of the tibia prosthetic coronal alignment accuracy between the tibia cutting without guide and the traditional extramedullary guided bone cutting by experienced surgeons. Only if when patients already have suffered severe malformation of knee joint (90-AT≥8°) before the operation, tibia cutting without cutting guide is more effective to rectify the tibia prosthetic coronal alignment.

Granny Smith apple procyanidin extract upregulates tight junction protein expression and modulates oxidative stress and inflammation in lipopolysaccharide-induced Caco-2 cells.

The present work is undertaken to characterize a Granny Smith apple procyanidin extract (AE) and investigate the beneficial effect of the AE in the intestine in vitro. Each AE was characterized via LC-ESI-MS. Caco-2 cells were used to study the preventive actions of the AE against the downregulation of tight junction protein expression, oxidative stress and inflammation induced by lipopolysaccharides (LPS). Phenolic compounds present in the AE, including chlorogenic acid, catechin, epicatechin, proanthocyanidin dimers, and proanthocyanidin trimers, were characterized. The expression of the tight junction protein, including occludin and zona occludens (ZO)-1, increased significantly in LPS + AE treated Caco-2 cells, compared to LPS induced Caco-2 cells. Proanthocyanidin dimers had the most potent effect on increasing tight junction protein expression. The addition of LPS to Caco-2 cells induced oxidative stress and inflammation. However, incubation with proanthocyanidin dimers prevented LPS-mediated oxidative stress, including the increase of SOD, HO-1, CAT, and GSH-Px mRNA expression, and counteracted LPS-mediated inflammation as evidenced by the down-regulation of inflammatory markers (NF-κß, IL-6, and TNF-α mRNA expression). Our findings provide evidence that AE could upregulate tight junction protein expression, probably acting via the reduction of oxidative stress and inflammation.

Bioinformatic and expression analysis of the OMT gene family in Pyrus bretschneideri cv. Dangshan Su.

With high nutritional value in its fruits, Dangshan Su pear has been widely cultivated in China. The stone cell content in fruits is a key factor affecting fruit quality in pear, and the formation of stone cells has been associated with lignin biosynthesis. O-Methyltransferase (OMT) is a key enzyme involved in lignin metabolism within the phenylpropanoid pathway. Here, we screened 26 OMT genes from the Pyrus bretschneideri cv. Dangshan Su genome using the DNATOOLs software. To characterize the OMT gene family in pear, gene structure, chromosomal localization, and conserved motifs of PbOMTs were analyzed. PbOMTs were divided into two categories, type I (designated PbCCOMTs) and type II (designated PbCOMTs), indicating the differentiation of function during evolution. Based on the analysis of multiple sequence alignment, cis-element prediction, and phylogenetic relationships, two candidate genes, PbCCOMT1 and PbCCOMT3, were selected for the analysis of temporal and spatial gene expression in pear. The promoter regions of both PbCCOMT1 and PbCCOMT3 contain regulatory motifs for lignin synthesis. Moreover, the two genes show high similarity and close phylogenetic relationships with CCOMTs in other species. Expression analysis showed that transcript levels of two PbCCOMTs were positively associated with the contents of both stone cells and lignin during the development of pear fruit. These results suggest that PbCCOMT1 and PbCCOMT3 are closely associated with lignin biosynthesis. These findings will help clarify the function of PbOMTs in lignin metabolism and to elucidate the mechanisms underlying stone cell formation in pear.

[Hydrogen sulfide in cartilage and its inhibitory effect on matrix metalloproteinase 13 expression in chondrocytes induced by interlukin-1ß].

OBJECTIVE: To investigate whether endogenous hydrogen sulfide (H2S) was involved in the pathogenesis of osteoarthritis (OA) and its underlying mechanism, to detect H2S and its synthases expression in knee cartilage in patients diagnosed with different severity of OA, and to explore the transcription and expression of gene MMP-13 in chondrocytes treated with IL-1ß or H2S. METHODS: Synovial fluids of the in-patients with different severity of OA hospitalized in Peking University First Hospital were collected for measurement of H2S content using methylene blue assay. Articular cartilages of the patients who underwent knee arthroplasty were collected for the cell culture of relatively normal chondrocytes. The chondrocytes were cultured to the P3 generation and H2S molecular probes were used for detection of endogenous H2S generation in the chondrocytes. Immunocytochemistry was used to detect the localization of H2S synthases including cystathionine ß-synthase (CBS), cystathionine-γ-lyase (CSE), and mercaptopyruvate sulfurtransferase (MPST) in OA chondrocytes. Western blot was used to quantify the protein expressions of CSE, MPST, and CBS in cartilage tissues of the patients who were diagnosed with OA and underwent knee arthroplasty. The relatively normal human chondrocytes were cultured to passage 3 and then divided into 4 groups for different treatments: (1)the normal control group, no reagent was added; (2)the IL-1ß group, 5 µg/L of IL-1ß was added; (3)the IL-1ß+H2S group, 200 µmol/L of NaHS was added 30 min before adding 5 µg/L of IL-1ß;(4)the H2S group, 200 µmol/L of NaHS was added. The transcription and expression of gene MMP-13 in chondrocytes of each group were determined with Real-time PCR and Western blot, respectively. And the total NF-κB p65 and phosphorylated NF-κB p65 in chondrocytes were detected with Western blot. RESULTS: The content of H2S in the synovial fluid of degenerative knee was (14.3±3.3) µmol/L. Expressions of endogenous H2S and its synthases including CBS, CSE and MPST were present in the cytoplasm of chondrocytes.CSE protein expression in Grade 3 (defined by outerbridge grading) cartilage tissues was significantly increased as compared with that of Grade 1 cartilage tissues (1.67±0.09 vs. 1.26±0.11, P< 0.05). However, no significant difference of CBS or MPST expression among the different groups was observed. The expression of MMP-13 protein in the IL-1ßgroup was significantly higher than that in the normal chondrocytes (1.87±0.67 vs. 0.22±0.10, P<0.05), and that in the IL-1ß+H2S group was significantly decreased than that in the IL-1ß group (0.55±0.11 vs. 1.87±0.67, P< 0.05), and that in the H2S group had no significant difference compared with that in the normal control group. The transcription of MMP-13 protein in the IL-1ß group was significantly higher than that in the normal chondrocytes (31.40±0.31 vs. 1.00±0.00, P<0.05), and that in the IL-1ß+H2S group was significantly decreased than that in the IL-1ß group (24.41±1.28 vs. 31.40±0.31, P<0.05), and that in the H2S group had no significant difference compared with that in the normal control group. The total NF-κB p65 in the IL-1ß group was significantly higher than that in the normal chondrocytes (2.13±0.08 vs. 0.73±0.08, P< 0.05), and that in the IL-1ß+H2S group was significantly decreased than that in the IL-1ß group (1.24±0.13 vs. 2.13±0.08, P<0.05), and that in the H2S group had no significant difference compared with that in the normal control group. The phosphorylated NF-κB p65 in IL-1ß group was significantly higher than that in the normal chondrocytes (1.30±0.13 vs. 0.19±0.04, P<0.05), and that in IL-1ß+H2S group was significantly decreased than that in the IL-1ß group (0.92±0.26 vs. 1.30±0.13, P<0.05), and that in the H2S group had no significant difference compared with that in the normal control group. CONCLUSION: H2S affected the cartilage degeneration by partly inhibiting the degradation of extracellular matrix.

Transthoracic closure of atrial septal defect and ventricular septal defect without cardiopulmonary bypass.

The minimally invasive surgical transthoracic occlusion of an atrial septal defect (ASD) or a ventricular septal defect (VSD) is an increasingly widespread alternative treatment for congenital heart disease. The aim of this study is to summarize our clinical experience with minimally invasive surgical transthoracic occlusion of ASD and VSD without cardiopulmonary bypass (CPB). Between April 2011 and October 2012, 27 patients with ASD and 95 patients with VSD (78 men and 44 women) were considered for minimally invasive surgical transthoracic occlusion without CPB. A small infrasternal incision (2.0-4.0 cm) was made under general anesthesia, under transesophageal echocardiography (TEE) guidance; the ASD and VSD were closed by using an appropriate occluder; and TEE was performed simultaneously to demonstrate the position of the device, any residual shunting, or encroachment on the atrioventricular valve, coronary sinus, or aortic valve. Successful transthoracic occlusion was performed in all 122 patients without complications. No complications such as third-degree atrioventricular block and residual shunting occurred after the procedures. The ventilation time was 2.2 ± 1.2 h, and the average length of hospital stay was 4.7 ± 1.7 days. All patients received aspirin at 3 mg·kg(-1)·day(-1) (maximum 100 mg/day) 24 h after the procedure. Minimally invasive surgical transthoracic occlusion without CPB is a new treatment that has many advantages such as causing little trauma, promoting quick recovery, having less complications, and avoiding radiation damage. However, the appropriate selection of patients is still key to improving the success rate of the operation.

[Bone marrow mesenchymal stem cells in Sprague-Dawley rat model of osteoarthritis].

OBJECTIVE: To investigate the efficacy of single time intra-articular different concentration of allogeneic bone marrow mesenchymal stem cells (BM-MSCs) injection in the treatment of Sprague-Dawley (SD) rat model of osteoarthritis (OA). METHODS: In the study, 32 SD rats were equally randomized into 4 groups: control group, high concentration group (1×10(7)/mL BM-MSCs), low concentration group (5×10(6)/mL BM-MSCs) and high vs. low concentration group. The two knees of each rat were set up to a pair. The induction of OA was performed surgically randomly at one side in model group, and bilaterally in the other groups, which were through anterior cruciate ligament transaction (ACLT) and medial meniscus excising. After the operation, the SD rats were allowed free movement. Four weeks later, different concentrations of allogeneic BM-MSCs isolated from the SD rats, expanded in vitro and suspended in phosphate buffered solution (PBS) were delivered in the articular cavity of both knees; PBS was used as the control. After injection, we excised the femoral nerve and sciatic nerve to disuse the low limb. The cartilage histological sections of knees were scored by Mankin scoring system to assess the severity of the pathology. mRNA of collagen II was detected by real time polymerase chain reaction (RT-PCR). eGFP was detected by fluorescence microscope. Assessments were carried out 4 weeks after the operation in model group, and 3 weeks after injection in the other groups. RESULTS: Mankin scores of the BM-MSCs side and control side were 6.60±0.40 vs. 10.00±0.32 in low concentration group (P<0.05), and 5.40±0.51 vs. 9.60 ±0.51 in high concentration group (P<0.05). Mankin scores of high vs. low concentration group were 6.40±0.51 vs. 7.60±0.75 (P>0.05). mRNA expression of collagen II of the BM-MSCs side in low concentration group was 106%±1% in contrast to the control side. As in high concentration group it was 108%±1%, and 102%±1% in high vs. low concentration group. Labeled BM-MSCs were detected unexpectedly in the synovial membrane but not in cartilage tissue three weeks from injection. CONCLUSION: BM-MSCs could promote cartilage repair and inhibit OA progression through a trophic mechanism. There was no difference between the two concentrations.

Single nucleotide polymorphisms on SHIP2 is associated with Type 2 diabetes mellitus in Chinese Han population.

OBJECTIVE: Type 2 diabetes mellitus (T2DM) is a chronic disease characterized by insulin resistance in the target tissue of insulin with insufficient insulin secretion in pancreatic ß-cells. Src homology 2-containing 5'-inositol phosphatase 2 (SHIP2) is a lipid phosphatase that hydrolyzes PI3-kinase product PI(3,4,5)P3 to PI(3,4)P2, which contributes to the negative regulation of insulin signaling both in vitro and in vivo. Some polymorphisms of SHIP2 have been reported to be associated with the metabolic syndrome including T2DM and hypertension in British, French and Japanese T2DM population. PATIENTS AND METHODS: In our present study, we investigated the relation between single nucleotide polymorphisms (SNPs) on SHIP2 gene and the pathogenesis of T2DM in Chinese Han population. RESULTS AND CONCLUSIONS: Our results indicated that the genotype and allele frequency of SHIP2 (+1893CC/AA) locus in T2DM patients showed significantly different from between the healthy control population. In addition, the G allele of SHIP2 (+2945A/G) seemed to increase the susceptibility to hypertension for T2DM patients.

Forced limb-use enhanced neurogenesis and behavioral recovery after stroke in the aged rats.

Constraint-induced movement therapy (CIMT) after stroke enhances not only functional reorganization but also structural plasticity of the brain in the adult rats. We examined whether forced limb-use which mimicked CIMT could influence ischemia-induced neurogenesis, apoptosis and behavioral recovery in the aged rats. Aged rats were divided into a sham group, an ischemia group, and an ischemia group with forced limb-use. Focal cerebral ischemia was induced by injection of endothelin-1. Forced limb-use began on post-stroke day 7 by fitting a plaster cast around the unimpaired upper limbs of rats for 3 weeks. Behavioral recovery was evaluated by tapered/ledged beam-walking test on postoperative day 32. The expression of doublecortin, neuronal nuclei, glial fibrillary acidic protein and Iba-1 were measured by single or double immunohistochemistry, and apoptosis was measured by TdT-mediated dUTP-biotin nick-end labeling (TUNEL) assay. The production of neuroblasts in the subventricular zone (SVZ) was significantly increased after stroke. Forced limb-use enhanced the proliferation of newborn neurons in the SVZ, as well as increased the long-term survival of newborn neurons. Furthermore, forced limb-use suppressed apoptosis and improved the motor functions after stroke in the aged rats. Forced limb-use exerted few effects on inflammation. Neither the number nor dendritic complexity of newborn granule cells in the hippocampus was affected by forced limb-use. Forced limb-use is effective in enhancing neurogenesis and behavioral recovery after stroke even in the aged rats.

Cloning and characterization of 5-enopyruvylshikimate-3-phosphate synthase from Pantoea sp.

The shikimate pathway enzyme 5-enopyruvylshikimate-3-phosphate synthase (EPSPS) is the target of the broad spectrum herbicide glyphosate. A novel aroA gene encoding an EPSPS from Pantoea sp was identified and subcloned into the pET-28a vector to construct the recombinant pET-AroAPantoea sp plasmid. Amino acid sequence analysis indicated that AroAPantoea sp is a class I AroA enzyme. When expressed in Escherichia coli, it conveyed high tolerance to glyphosate. AroAPantoea sp may be used to generate transgenic glyphosate-tolerant plants.

All-trans retinoic acid induces cell-cycle arrest in human cutaneous squamous carcinoma cells by inhibiting the mitogen-activated protein kinase-activated protein 1 pathway.

BACKGROUND: All-trans retinoic acid (ATRA) has been tried for the treatment and prevention of a number of epithelial cancers. However, the precise mechanism by which ATRA inhibits the growth of cutaneous squamous cell carcinoma (cSCC) remains elusive. AIMS: To determine the suppressive effects of ATRA on the human cSCC cell line SCL-1, and explore the possible mechanisms involved. METHODS: SCL-1 cells were treated with ATRA, then cell proliferation was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, while apoptosis and cell cycle progression were analysed by flow cytometry. Protein levels of cell-cycle regulatory proteins and the activation of extracellular signal-regulated kinase (ERK) and Jun kinase (JNK) were detected by western blotting analysis. Transcriptional activity of activator protein (AP)-1 was examined by luciferase reporter assay. RESULTS: ATRA inhibited the proliferation of SCL-1 cells and had modest proapoptotic effects. ATRA also induced G1 cell-cycle arrest, inhibited the expression of cyclin D1/cyclin-dependent kinase (CDK)4 and cyclinE/CDK2, and increased the expression of the cyclin-dependent kinase inhibitors p21 and p27. In addition, ATRA significantly decreased the phosphorylation of ERK1/2 and JNK1/2, and inhibited AP-1 transcriptional activity. CONCLUSIONS: ATRA induces cell-cycle arrest in human cSCC cells by inhibiting the mitogen-activated protein kinase (MAPK)-AP1 pathway, and could be effective in the prevention and chemotherapy of human cSCC.

Deletion of estrogen receptor beta accelerates early stage of bone healing in a mouse osteotomy model.

UNLABELLED: This study examined the role of estrogen receptor (ER) beta during mouse femoral fracture healing by employing ER knockout (KO) mice. The fracture healing in KO mice was enhanced in the early stage of neovascularization and the middle stage of endochondral ossification. INTRODUCTION: This study was conducted to examine the role of ER beta during fracture healing. METHODS: Female ERbeta knockout (KO) mice (18 weeks old) and age-matched female wild-type (WT) mice underwent open osteotomy on the right femur. They were sacrificed at 1, 2, 4 and 6 weeks post-fracture. The sera and callus samples were subjected to the following analyses: micro-computed tomography (CT)-based angiography, micro-CT evaluation, histological examination, histomorphometry examination, real-time polymerase chain reaction (PCR) analysis, biochemical marker, and mechanical testing. RESULTS: Micro-CT-based angiography showed that the total vessel volume at the fracture site was larger in the KO group than the WT group at 1 and 2 weeks post-fracture. Micro-CT analysis revealed that the callus volume was significantly higher in the KO group from week 2 to week 4 post-fracture when compared with the WT group consistent with the histological data. Analysis of biochemical markers indicated that circulating P1NP levels in the KO mice were significantly higher than in the WT mice from week 2 to week 4 and that temporal expression of circulating C-terminal telopeptide of type I collagen (CTX) levels was also higher in the KO mice than in the WT mice. These results were consistent with quantitative real-time PCR analysis. The ultimate load, stiffness, and energy to failure were significantly higher in the KO mice than in the WT mice at week 4. CONCLUSIONS: The fracture healing in KO mice was enhanced in the early stage of neovascularization and the middle stage of endochondral ossification, but not by the end of healing. Blockade of ERbeta can be considered as another therapeutic strategy for osteoporotic fracture and non-union fracture.

[Unusual manifestation of cerebral cysticercosis].

We have surgically treated two patients with cerebral cysticercosis, which were pre-operatively unexpected due to unusual CT and MRI manifestations. A 24 year old chinese farmer with intermittent severe headache for 6 years was examined by CT and MRI. A huge cystic lesion with same density or intensities to CSF occupied retroclival posterior fossa to upper cervical spinal canal and displaced neural structures backwardly. By suboccipital craniectomy, watery clear cyst fluid of 44ml was aspirated and the shrunken cyst was pulled out with ease. It was unable to predict until an electron microscopic study, which revealed a larva with three layers of microvilli, vesicular tegmentum and infrategmental muscle bundles and vesicles. The findings were same to those of racemose cysticercus of human brain. Giant cysts of cysticercosis were infrequently present supratentorially, although infratentorial or basal cysticercosis were racemose and small. The present case may be the first case of a huge cyst of cysticercosis in posterior fossa. A 44 year old Japanese businessman with 15 years history of general convulsion underwent craniotomy due to a small, but growing cystic granulomatous mass in right inferior frontal gyrus. The CT scan examined at 3 years after his first general convulsion revealed a small round low density mass, 5mm in diameter, in right frontal lobe and its wall was weakly enhanced by contrast media. The follow-up CT scans and MRIs revealed that the enhancing cystic mass became thick and deformed, and perifocal edema came to evident and progressed stepwise by attacks of general convulsions.(ABSTRACT TRUNCATED AT 250 WORDS)