Three-dimensional Reconstruction and Comparison of Temporomandibular Joint Space Volume Before and After Orthognathic Surgery in Patients with Skeletal Class III Malocclusion With Mandibular Deviation.

OBJECTIVE: To look into the association between the degree of deviation and the changing trend in the temporomandibular joint (TMJ) space volume after orthognathic surgery in patients with skeletal Class III malocclusion. METHODS: Twenty patients having combined orthodontic-orthognathic treatment for skeletal Class III malocclusions with mandibular deviation were chosen, and craniofacial spiral CT was performed before (T0), two weeks after (T1), and six months after (T2) surgery. Using 3D volume reconstruction, further partitioning, and analysis of each domain's volume changes over time, the TMJ space volume is to be obtained. The differences in changes between groups A (mild deviation group) and B (severe deviation group) were examined to examine the impact of the degree of deviation on the TMJ space volume. RESULTS: A statistically significant difference ( P <0.05) existed between the postoperative TMJ space volume in group A and the preoperative overall, anterolateral, and anteroinferior space volume; the same difference also existed between the postoperative TMJ space volume in the NDS and the preoperative posterolateral, posteroinferior space volume. In group B, the postoperative TMJ space volume was statistically significant ( P <0.05) compared with the preoperative total and anteroinferior space volume in the DS; the difference between the total volume of the T1 stage on the NDS and the total volume of the T0 stage was statistically significant ( P <0.05). The two groups showed substantial differences in the space volume changes between the T1-T0 phase and the T2-T1 period. CONCLUSION: Patients with skeletal Class III malocclusion and mandibular deviation after orthognathic surgery see a change in the TMJ space volume. All patient types experience a largely consistent space volume change trend two weeks after surgery, and the degree of mandibular deviation is correlated with the intensity and longevity of the alteration.

ZEB1 Mediates Bone Marrow Mesenchymal Stem Cell Osteogenic Differentiation Partly via Wnt/ß-Catenin Signaling.

Zinc finger E-box-binding homebox 1 (ZEB1) is a zinc-finger transcription factor best known for its role in promoting the epithelial-mesenchymal transition, which is also related to osteogenesis. Here, ZEB1 was investigated for its role in the commitment of bone marrow mesenchymal stem cells (BMSCs) to osteoblasts. In vitro, ZEB1 expression decreased following osteogenic differentiation. Furthermore, silencing of ZEB1 in BMSCs promoted osteogenic activity and mineralization. The increase in osteogenic differentiation induced by si-ZEB1 could be partly rescued by the inhibition of Wnt/ß-catenin (si-ß-catenin). In vivo, knockdown of ZEB1 in BMSCs inhibited the rapid bone loss of ovariectomized (OVX) mice. ZEB1 expression has also been negatively associated with bone mass and bone formation in postmenopausal women. In conclusion, ZEB1 is an essential transcription factor in BMSC differentiation and may serve as a potential anabolic strategy for treating and preventing postmenopausal osteoporosis (PMOP).

Oat-Derived ß-Glucans Induced Trained Immunity Through Metabolic Reprogramming.

Trained immunity has been recently identified in innate immune cells, which undergo long-term epigenetic and metabolic reprogramming after exposure to pathogens for protection from secondary infections. (1, 3)/(1, 6)-ß-glucan derived from fungi can induce potent trained immunity; however, the effect of (1, 3)/(1, 4)-ß-glucan (rich in dietary fiber oat) on trained immunity has not been reported. In the present study, two cell culture systems for trained immunity induction were validated in monocytes/macrophages from mouse bone myeloid and human THP-1 cells exposed to positive inducers of trained immunity, including ß-glucan from Trametes versicolor or human-oxidized low-density lipoprotein. Primed with oat ß-glucan, the mRNA expression and production of TNF-α and IL-6 significantly increased in response to re-stimulation of TLR-4/2 ligands. Moreover, the expression of several key enzymes in glycolytic pathway and tricarboxylic acid cycle was significantly upregulated. In addition, inhibiting these enzymes decreased the production of TNF-α and IL-6 boosted by oat ß-glucan. These results show that oat ß-glucan induces trained immunity through metabolic reprogramming. This provides important evidence that dietary fiber can maintain the long-term responsiveness of the innate immune system, which may benefit for prevention of infectious diseases or cancers.

PTH1-34 improves bone healing by promoting angiogenesis and facilitating MSCs migration and differentiation in a stabilized fracture mouse model.

OBJECTIVE: PTH1-34 (parathyroid hormone 1-34) is the only clinical drug to promote osteogenesis. MSCs (mesenchymal stem cells) have multidirectional differentiation potential and are closely related to fracture healing. This study was to explore the effects of PTH1-34 on proliferation and differentiation of endothelial cells and MSCs in vitro, and on angiogenesis, and MSCs migration during fracture healing in vivo. METHODS: Mice with stabilized fracture were assigned to 4 groups: CON, PTH (PTH1-34 40 µg/kg/day), MSC (transplanted with 105 MSCs), PTH+MSCs. Mice were sacrificed 14 days after fracture, and callus tissues were harvested for microCT scan and immunohistochemistry analysis. The effects of PTH1-34 on angiogenesis, and MSCs differentiation and migration were assessed by wound healing, tube formation and immunofluorescence staining. RESULTS: Treatment with either PTH1-34, or MSCs promoted bone healing and vascular formation in fracture callus. The callus bone mass, bone volume, and bone mineral density were all greater in PTH and/or MSC groups than they were in CON (p<0.05). PTH1-34 increased small vessels formation (diameter ≤50µm), whereas MSCs increased the large ones (diameter >50µm). Expression of CD31 within calluses and trabecular bones were significantly higher in PTH1-34 treated group than that of not (p<0.05). Expression of CD31, VEGFR, VEGFR2, and vWF was upregulated, and wound healing and tube formation were increased in MSCs treated with PTH1-34 compared to that of control. CONCLUSIONS: PTH1-34 improved the proliferation and differentiation of endothelial cells and MSCs, enhancing migration of MSCs to bone callus to promote angiogenesis and osteogenesis, and facilitating fracture healing.

Development and validation of a radiomics nomogram for identifying invasiveness of pulmonary adenocarcinomas appearing as subcentimeter ground-glass opacity nodules.

The aim of the present study was to develop and validate a radiomics-based nomogram for differentiation of pre-invasive lesions from invasive lesions that appearing as ground-glass opacity nodules (GGNs) ≤10 mm (sub-centimeter) in diameter at CT. A total of 542 consecutive patients with 626 pathologically confirmed pulmonary subcentimeter GGNs were retrospectively studied from October 2011 to September 2017. All the GGNs were divided into a training set (n = 334) and a validation set (n = 292). Researchers extracted 475 radiomics features from the plain CT images; a radiomics signature was constructed with the least absolute shrinkage and selection operator (LASSO) based on multivariable regression in the training set. Based on the multivariable logistic regression model, a radiomics nomogram was developed in the training set. The performance of the nomogram was evaluated with respect to its calibration, discrimination, and clinical-utility and this was assessed in the validation set. The constructed radiomics signature, which consisted of 15 radiomics features, was significantly associated with the invasiveness of subcentimeter GGNs (P < 0.0001 for both training set and validation set). To build the nomogram model, radiomics signature and mean CT value were used. The nomogram model demonstrated good discrimination and calibration in both training set (C-index, 0.716 [95% CI, 0.632 to 0.801]) and validation set (C-index, 0.707 [95% CI, 0.625 to 0.788]). Decision curve analysis (DCA) indicated that radiomics-based nomogram was clinically useful. A radiomics-based nomogram that incorporates both radiomics signature and mean CT value is constructed in the study, which can be conveniently used to facilitate the preoperative individualized prediction of the invasiveness in patients with subcentimeter GGNs.

Self-Replication-Assisted Rapid Preparation of DNA Nanowires at Room Temperature and Its Biosensing Application.

A rapid room-temperature DNA nanowires preparation strategy on the basis of self-replicating catalyzed hairpin assembly (SRCHA) was reported. In this system, three hairpin probes (P1, P2, and P3) were well-designed and partially hybridize to each other, and two split trigger DNA sequences were integrated into P1 and P3, respectively. When the SRCHA was initiated by the trigger DNA, a series of DNA assembly steps based on the toehold-mediated DNA strand displacement were activated, and the Y shaped DNA (P1-P2-P3) was formed. In that case, the two split trigger DNA sequences will come into close-enough proximity to form the trigger DNA replicas, which can initiate the additional SRCHA reaction cycles for DNA nanowire preparation, and eventually a rapid room-temperature DNA nanowires preparation strategy without need of fuel strands was successfully developed. Furthermore, the prepared DNA nanowires have been used to develop a rapid and signal amplified sensing platform for sensitive adenosine triphosphate (ATP) detection.

Management of prolactinoma: a survey of endocrinologists in China.

Objective To assess the current management of prolactinoma among endocrinologists in China. Methods An online survey of a large sample of endocrinologists was conducted in China. The questionnaire included 21 questions related to controversial issues about the management of prolactinomas. Doctors in the endocrinology department of a university-affiliated hospital or a comprehensive secondary hospital in 12 cities from East, West, South, North and Middle China were surveyed. Results A total of 290 valid questionnaires were collected, and the response rate was 40%. When hyperprolactinemia occurred, 97% of the respondents would test thyroid-stimulating hormone routinely. 22% of the respondents considered that prolactin levels <100 ng/mL exclude the presence of a prolactinoma. Only 9% of the respondents believed that prolactin >250 ng/mL could occur in all the following situations as macroprolactinoma, mircoprolactinoma, macroprolactinemia and drug-induced hyperprolactinemia. Surgery was not recommended by 272 (94%) endocrinologists as the first choice for treating microprolactinomas. 58% and 92% of endocrinologists would start drug treatment for microprolactinomas and macroprolactinomas at diagnosis. 70% and 40% chose to withdraw treatment after 2-3 years of prolactin normalization in microprolactinomas and macroprolactinomas. In case of pregnancy, 57% of the respondents considered bromocriptine as choice for women patients. Drug discontinuation after pregnancy was advocated in 63% and 27% for microprolactinoma and macroprolactinoma. Moreover, 44% of endocrinologists believed that breastfeeding was allowable in both micro- and macroprolactinoma. Conclusion This is the first study to investigate the management of prolactinomas among endocrinologists in China. We found that the current clinical treatment was not uniform. Therefore, it is necessary to strengthen the training of endocrinologists to improve clinical diagnosis and treatment practices.

Signs analysis and clinical assessment: phase-contrast computed tomography of human breast tumours.

PURPOSE: To analyse the diagnostic signs present in slices of human breast tumour specimens using synchrotron radiation phase-contrast imaging computed tomography (PCI-CT) for the first time and assess the feasibility of this technique for clinical applications. MATERIALS AND METHODS: The ethics committee of our university and relevant clinical hospital approved this prospective study, and written informed consent was obtained from all patients. PCI-CT of human breast tumour specimens with synchrotron radiation was performed at the Shanghai Synchrotron Radiation Facility (SSRF). A total of 14 specimens of early-stage carcinomas and 8 specimens of adenomas were enrolled. Based on raw data reconstruction, the diagnostic signs present in the slices were analysed and correlated with histopathology. We proposed a criterion for clinical diagnosis according to the evaluated signs and the Breast Imaging Reporting and Data System (BI-RADS) for reference. The criterion was then assessed by clinicians in a double-blind method. Finally, descriptive statistics were evaluated, depending on the assessment results. RESULTS: The 14 carcinoma specimens and 8 adenoma specimens were diagnosed as malignant and benign tumours, respectively. The total coincidence rate was 100%. CONCLUSION: Our study results demonstrate that the X-ray diagnostic signs observed in the specimen slices and the criterion used for clinical diagnosis were accurate and reliable. The criterion based on signs analysis can be used to differentiate early-stage benign or malignant tumours. As a promising imaging method, PCI-CT can serve as a possible and feasible supplement to BI-RADS in the future.

Peripheral neuropathy is associated with insulin resistance independent of metabolic syndrome.

BACKGROUND: To determine the association of insulin resistance, metabolic syndrome (MetS) with peripheral neuropathy (PN). METHODS: This cross-sectional study consisted of 2035 subjects in Shanghai who were classified as with MetS and without MetS. The new International Diabetes Federation (IDF) criterion was used to define MetS. HOMA-IR was applied to evaluate insulin resistance. All subjects underwent complete foot examination. PN was assessed according to the neuropathy symptom and neuropathy disability scores. Binary logistic regression was performed to analyze the contributions of insulin resistance, features of MetS to PN. RESULTS: (1) The percentage of PN was 4.0% in our study. Patients with MetS (47.7%) had a higher percentage of PN (5.5% vs. 2.6%, respectively, P = 0.001). With the components of MetS increased (non-MetS, three, four, five), a linear increase in the proportion of peripheral neuropathy was observed (2.6%, 4.8%, 5.6% and 7.2%; respectively, P for trend = 0.001). (2) In patients with PN, the average age of patients was significantly older than the corresponding non-PN patients. Waist circumference, fasting blood glucose, HbA1c, proportion of treatment for diabetes and hypertension were significantly higher in PN group compared with non-PN group in MetS patients. (3) The frequency of dysglycemia was the highest in PN patients both with and without MetS (96.2% and 82.1%, P = 0.084). (4) After adjusting for gender and smoking history, the PN was associated with MetS [odds ratio (OR) 2.0; 95% confidence interval (CI) 1.2, 3.2; P = 0.006], and age (OR 1.1; 95% CI 1.1, 1.1; P < 0.001). When HOMA-IR was added to this binary logistic regression, the association of PN with MetS disappeared (P = 0.110), but the PN was still associated with HOMA-IR (OR 1.2; 95% CI 1.1, 1.4, P < 0.001). CONCLUSIONS: In metabolic syndrome, insulin resistance might play an important role in the development of peripheral neuropathy.

Purification of a Pd20-TNFα fusion protein that prevents liver metastasis of gastric cancer.

The specific binding peptide pd20 of gastric cancer cells with a high potential for liver metastasis was fused with human tumour necrosis factor (TNF) α, and a prokaryotic expression vector was established to express the pd20-TNFα fusion protein. After purification and identification, the preventive effects of the fusion protein on liver metastasis of gastric cancer were observed in mice. The whole gene synthesis method was used for pd20-TNFα fusion gene preparation, and a pd20-TNFα prokaryotic expression vector was constructed. The vector was induced and expressed in Escherichia coli BL21. The expression products were analysed and verified by SDS-PAGE electrophoresis and Western blot analysis. The Ni-NTA column method was used to purify the fusion protein, and the L929 cytotoxicity method was used to detect biological activity. Flow cytometry apoptosis experiments and invasion assays were performed to observe the effects of the fusion protein on apoptosis and metastasis of gastric cancer cells with high potential for liver metastasis. Thirty nude mice with liver metastasis of gastric cancer were established and then randomly divided into three groups of ten mice each. The Pd20-TNFα recombinant protein (1.2 × 10(6) U/kg day) or standard TNFα (1.2 × 10(6) U/kg day) saline was administered via tail vein injection for 7 consecutive days. The pathological changes in various organs of nude mice were observed 4 weeks later. The size of the gastric cancer, the incidence of liver metastasis and the number of liver metastases were measured and calculated. We successfully constructed a Pd20-TNFα recombinant plasmid and prepared the fusion protein. Detection of the pd20-TNFα protein by immunofluorescence showed a very strong expression in liver tissue, suggesting a targeting of the fusion protein to the liver. The L929 cytotoxicity assays showed that the pd20-TNFα fusion purified protein had a significant lethal effect on L929 cells, with a killing activity of up to 7.6 × 10(6) IU/ml. The apoptosis experiments showed that as the concentration of the fusion protein increased, the early gastric cancer cell apoptosis also increased, with the early apoptosis rate increasing from 5.99 % to 9.04 %. Cell invasion experiments showed that the purified pd20-TNFα fusion protein significantly inhibited the in vitro invasion of XGC9811-L cells, with the penetrating cells being significantly decreased compared with the control group per unit time (P < 0.01). Vector experiments showed that the pd20-TNFα recombinant protein group had significantly reduced cancer lesions and liver metastasis in nude mice compared with the control group. We successfully purified a pd20-TNFα fusion protein and confirmed that it had significant biological activity promoting early gastric cancer cell apoptosis, thereby inhibiting gastric cancer cell invasion.

[Preliminary study on effect of surgical delay on anti-infection ability of reverse fascio-cutaneous flap].

OBJECTIVE: To improve the success rate of the reverse fascio-cutaneous flap in repairing the infected wound, to observe the effect of surgical delay on the anti-infection ability of the reverse fascio-cutaneous flap by establishing an oryctolagus cuniculus model of reverse fascio-cutaneous flap based on sural nerve on the lateral side of left later limb. METHODS: Sixteen 5-month-old Japanese white rabbits weighing 2.0-2.5 kg (mean, 2.3 kg) were randomly divided into experimental group (n = 8) and control group (n = 8). The reverse fascio-cutaneous flap of 4 cm x 2 cm was designed, based on 1 cm above the lateral malleolar as pedicle in sural nerve region in the lateral left later limb. In the experimental group, the full-thickness of the flap distal end half was harvested according to the design; and after 10 days delay, the full-thickness flap was obtained according to the design, and 0.5 mL Staphylococcus aureus solution was implanted at a density of 3.8 x 10(6)/mL in 2 groups. The general observation was performed postoperatively; the venous blood of the marginal ear vein was collected to observe white blood cell (WBC) count before implantation of staphylococcus aureus solution and after 1, 3, 5, 7, 10, and 14 days of implantation. The flap survival rate and the colony counting of necrosis flap tissue were calculated after 10 days of implantation; the blood vessel caliber and the peak value of peroneal artery blood flow of flap proximal end were measured after 14 days of implantation. RESULTS: All animals survived to the end of the experiment, and all incisions healed primarily. Inflammatory reaction with different degrees was observed after implantation in 2 groups, and it was obvious at 3-5 days. Inflammatory reaction in the experimental group was slighter than that in the control group. Except for no significant difference before implantation and after 14 days of implantation between 2 groups (P > 0.05), there were significant differences in WBC count at other time points between 2 groups (P < 0.05). The flap survival rate of the experimental group (93.20% +/- 4.62%) was significant higher than that of the control group (72.65% +/- 7.80%) after 10 days of implantation (P < 0.05). The colony counting of necrosis flap tissue in the experimental group [(20.63 +/- 5.76) x 10(3) colony/g] was significantly lower than that in the control group [(32.38 +/- 6.14) x 10(3) colony/g] after 10 days of implantation (P < 0.05). The blood vessel caliber of the experimental group and the control group were (1.03 +/- 0.10) mm and (0.75 +/- 0.09) mm, respectively, and the peak value of peroneal artery blood flow in the experimental group and the control group were (20.73 +/- 2.46) cm/s and (13.83 +/- 1.51) cm/s, respectively, after 14 days of implantation; showing significant differences between 2 groups (P < 0.05). CONCLUSION: Surgical delay has the ability of enhancing survival and anti-infection of the rabbit reverse fascio-cutaneous flap.

[Effect of hepatitis B virus X gene on the apoptosis of HepG2 cells].

OBJECTIVE: To investigate the effect of hepatitis B virus X gene on the apoptosis in X gene-transfected HepG2 cells. METHODS: HBX gene eukaryon expression vector pcDNA3.1-X was transfected into HepG2 cells by lipid-mediated transfection to establish HepG2/HBX cell model for HBX expression. HepG2 transfected with pcDNA3.1 was used as controls. At 24 h, 48 h, 72 h and 96 h after transfection, cell apoptotic rates were detected by flow cytometry. RT-PCR and Western blot were applied to evaluate the expression levels of HBX, Fas, FasL, and the levels of p-JNK and p-c-Jun protein. RESULTS: HBX mRNA was detected in HepG2/ HBX cells 24 h after transfection, and the expression of HBX mRNA level was gradually up-regulated after transfection (P < 0.05); whereas no expression of HBX gene in the control cells. At 24 h, 48 h, 72 h and 96 h after transfection, all of the apoptotic rate in HepG2/HBX group were much higher than those in the controls (P < 0.05). The expression levels of Fas and FasL protein as well as the levels of p-JNK and p-c-Jun protein were significantly higher than those in the controls (P < 0.05). A positive correlationship was observed between the expression of HBX mRNA level and the hepatocyte apoptotic rate (P < 0.05), the same correlation of HBX mRNA level with the levels of Fas, FasL, p-JNK and p-c-Jun were also observed. (P < 0.05). CONCLUSION: HBX can induce hepatocyte apoptosis by up-regulating the levels of p-JNK and p-c-Jun to promote the expression of FasL.