PVH-Peri5 Pathway for Stress-Coping Oromotor and Anxious Behaviors in Mice.

Stressful stimuli can activate the hypothalamic-pituitary-adrenal (HPA) axis. Clinically, it has been widely reported that stressful events are often accompanied by teeth clenching and bruxism, while mastication (chewing) can promote coping with stress. Trigeminal motoneurons in the trigeminal motor nucleus supplying the chewing muscles receive direct inputs from interneurons within the peritrigeminal premotor area (Peri5). Previous studies found that the paraventricular hypothalamic nucleus (PVH) participates in trigeminal activities during stressful events. However, the neural pathway by which the stress-induced oral movements alleviate stress is largely unknown. We hypothesized that paraventricular-trigeminal circuits might be associated with the stress-induced chewing movements and anxiety levels. First, we observed the stress-coping effect of wood gnawing on stress-induced anxiety, with less anxiety-like behaviors seen in the open field test and elevated plus maze, as well as decreased corticosterone and blood glucose levels, in response to stress in mice. We then found that excitotoxic lesions of PVH reduced the effect of gnawing on stress, reflected in more anxiety-like behaviors; this emphasizes the importance of the PVH in stress responses. Anterograde, retrograde, transsynaptic, and nontranssynaptic tracing through central and peripheral injections confirmed monosynaptic projections from PVH to Peri5. We discovered that PVH receives proprioceptive sensory inputs from the jaw muscle and periodontal ligaments, as well as provides motor outputs via the mesencephalic trigeminal nucleus (Me5) and Peri5. Next, pathway-specific functional manipulation by chemogenetic inhibition was conducted to further explore the role of PVH-Peri5 monosynaptic projections. Remarkably, PVH-Peri5 inhibition decreased gnawing but did not necessarily reduce stress-induced anxiety. Moreover, neuropeptide B (NPB) was expressed in Peri5-projecting PVH neurons, indicating that NPB signaling may mediate the effects of PVH-Peri5. In conclusion, our data revealed a PVH-Peri5 circuit that plays a role in the stress response via its associations with oromotor movements and relative anxiety-like behaviors.

[Influences of leukocytes in patients with type 2 diabetes and periodontitis to the effects of periodontal treatment on glycemic control].

Objective: To analyze the influences of leukocytes on improving blood glucose control in patients with type 2 diabetes mellitus (T2DM) and periodontitis after periodontal mechanical therapy. Methods: Thirty-five patients visiting Peking University Third Hospital from March 2011 to August 2012, as well as thirty-four patients visiting Peking University Hospital of Stomatology from March 2011 to August 2012 and December 2016 to December 2018 were selected in this research. These subjects were non-smokers, and with moderate to severe chronic periodontitis and T2DM. The full set of periodontal examinations including probing depth (PD), attachment loss (AL), bleeding index (BI) and plaque index (PLI) were conducted. Besides, counts of white blood cells (WBC), parameters of glucose and lipids metabolites such as fasting blood glucose (FBG), glycosylated hemoglobin (HbA1c), total cholesterol (TC), triglyceride (TG), high density lipoprotein (HDL) and low density lipoprotein (LDL) in serum were examined before treatment. Then, oral hygiene instruction, scaling and root planing (SRP) were carried out. Three months after SRP, the baseline examinations were repeated in all patients. According to the baseline leukocyte counts, the patients were divided into subgroups: low WBC group (WBC<6.19×109/L) and high WBC group (WBC≥6.19×109/L). Paired t-test for comparison of changes after treatment, analysis of co-variance for comparing the intervention effects between subgroups, and multifactor Iogistic regression analysis were performed. Results: Three months after SRP, all periodontal indexes were significantly improved in both groups. Leukocyte counts decreased significantly in high WBC group (7.64±1.51 vs. 6.89±1.53, P=0.008). In high WBC group, HbA1c (7.67±1.35 vs. 7.18±1.09, P=0.001) and LDL (3.28±0.76 vs. 2.67±0.85, P=0.042) decreased significantly, while there were no such differences in low WBC group. Influence of leukocyte level on HbA1c (OR=0.12, P=0.038) and LDL (OR=0.15, P=0.001) improvement was statistically significant. Hierarchical analysis showed such improvement notably perform in female [HbA1c (OR=0.30, P=0.021), LDL (OR=0.34, P=0.001)] and severe periodontitis group [HbA1c (OR=0.15, P=0.025), LDL (OR=0.24, P=0.017)]. Through interaction test, female and leukocyte counts at baseline had relative excess risk affecting the effect of periodontal intervention on HbA1c (P=0.036) and LDL (P=0.005). Conclusions: SRP could significantly improve the blood glucose and lipid control in patients who had T2DM and chronic periodontitis with relative higher leukocytes level. Female patients with severe periodontitis showed more obviously effects.

Effect of Al substitution on the magnetization reversal and complex magnetic properties of NiCr2O4 ceramics.

A series of polycrystalline NiCr2-xAlxO4 (0.1 ≤ x ≤ 0.25) spinel ceramics have been synthesized using a sol-gel method. DC magnetization measurements are carried out at different temperatures and magnetic fields. A novel magnetization reversal has been observed in the field cooling process for the x = 0.2 sample, which can be ascribed to the competition between two magnetic sublattices due to their different temperature dependences. The magnetic interaction evolution, related to the complex magnetic properties, is revealed by exchange constants that have been estimated according to ferrimagnetic Curie-Weiss fitting and mean field theory. The fitting result confirmed the evolution of antiferromagnetic and ferromagnetic components with Al substitution, which is supported by the observations from the isothermal magnetization measurements. The positive and negative values of the magnetic moment can be utilized for storage applications based on the results of magnetic switching effect measurements.

[Effects of LINC00839 targeting miR-3666 on proliferation, migration and invasion of hepatocellular carcinoma cells].

Objective: To investigate the effects of lncRNA LINC00839 on the proliferation, migration and invasion of hepatocellular carcinoma cells and its mechanism. Methods: Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of LINC00839 and miR-3666 in hepatocellular carcinoma tissues and adjacent tissues. Pearson correlation was used to analyze the correlation between LINC00839 and miR-3666 expression in liver cancer tissues. Hepatocellular carcinoma cells MHCC97H were cultured in vitro and divided into si-NC group, si-LINC00839 group, miR-NC group, miR-3666 group, si-LINC00839+ anti-miR-NC group, and si-LINC00839+ anti-miR-3666 group. Methylthiazoletrazolium (MTT) method and clone formation experiment were used to detect cell proliferation. Transwell array was used to detect the cell migration and invasion. Western blot was used to detect the protein expressions of p21, E-cadherin and MMP-2. The double luciferase reporter gene experiment was used to verify the regulatory relationship between LINC00839 and miR-3666. Results: Compared with adjacent tissues, the expression level of LINC00839 in hepatocellular carcinoma tissues increased (2.82±0.27 vs. 0.96±0.10, P<0.001), but the expression level of miR-3666 decreased (0.23±0.02 vs. 1.01±0.10, P<0.001). The expression levels of LINC00839 and miR-3666 in liver cancer tissue were negatively correlated (r=-0.658, P<0.001). The survival rate of MHCC97H cells in the si-LINC00839 group [(53.91±5.41)% vs. (100.53±10.22)%], the number of clones formed (92.0±8.0 vs. 164.0±14.3), the number of migration (131.0±12.7 vs. 247.0±22.4), the number of invasion (66.0±6.4 vs. 120.0±11.6) and the protein level of MMP-2 (0.20±0.02 vs. 0.67±0.06) were lower than those in the si-NC group (P<0.001). However, the protein levels of p21 (0.76±0.07 vs. 0.25±0.02) and E-cadherin (0.78±0.08 vs. 0.14±0.01) were higher than those in the si-NC group (P<0.001). LINC00839 targeted and negatively regulated the expression of miR-3666. The survival rate of MHCC97-H cells in the miR-3666 group [(47.93±4.86)% vs. (100.11±10.21)%], the number of clone formation (78.0±7.7 vs. 166.0±15.9), the number of migration (117.0±12.1 vs. 250.0±25.0), the number of invasion (57.0±5.7 vs. 121.0±12.3) and the protein level of MMP-2 (0.16±0.01 vs. 0.69±0.07) were lower than those in the miR-NC group (all P<0.001). However, the protein levels of p21 (0.83±0.08 vs. 0.24±0.02) and E-cadherin (0.87±0.09 vs. 0.13±0.01)were higher than those in the miR-NC group (all P<0.001). The survival rate of MHCC97-H cells in the si-LINC00839+ anti-miR-3666 group [(89.94±9.05)% vs. (54.12±5.39)%], the number of clones (143.0±13.8 vs. 94.0±9.4), the number of migration (208.0±19.8 vs. 129.0±12.6), the number of invasion (108.0±10.1 vs. 65.0±6.4) and the protein level of MMP-2 (0.31±0.03 vs 0.66±0.06) were higher than those in the si-LINC00839+ anti-miR-NC group (P<0.001). However, the protein levels of p21 (0.31±0.03 vs. 0.74±0.07) and E-cadherin (0.28±0.03 vs. 0.80±0.08) were lower than those int the si-LINC00839+ anti-miR-NC group (P<0.001). Conclusion: Inhibition of LINC00839 expression may inhibit the proliferation, migration and invasion of hepatocellular carcinoma cells by targeting up-regulation of miR-3666 expression.

[Research of prognostic immunophenotypes in 163 patients of diffuse large B-cell lymphoma].

Objective: To screen and analyze the prognostic protein biomarkers of DLBCL, and to explore their value in the prognostic evaluation. Methods: 163 cases of confirmed DLBCLs from January 2011 to December 2016 were collected with their clinical, pathological and follow-up data, which were all from our hospital. The expression of protein markers were tested using immunohistochemical staining (IHC) . The immune phenotypes independent of the International Prognostic Index (IPI) that affect overall survival (OS) and progression-free survival (PFS) of DLBCL were explored by COX regression model, and the effect of their co-expression on the prognosis were also analyzed. Result: BCL6 negative (PFS: HR=1.652, 95%CI 1.030-2.649, P=0.037) , P53 positive (OS: HR=1.842, 95%CI 1.008-3.367, P=0.047) , and BCL2 strong positive expressions (S+) (OS: HR=2.102, 95%CI 1.249-3.537, P=0.005; PFS: HR=2.126, 95%CI 1.312-3.443, P=0.002) are adverse prognostic factors of DLBCL that are independent of IPI. BCL6(-) (PFS: HR=2.042, 95%CI 1.021-4.081, P=0.043) , P53(+) (OS: HR=3.069, 95%CI 1.244-7.569, P=0.015) and BCL2(S+) (OS: HR=2.433, 95%CI 1.165-5.082, P=0.018; PFS: HR=3.209, 95%CI 1.606-6.410, P=0.001) are adverse prognostic factors in the group of age≤60-year-old; in the group of IPI score 0-2, cases with BCL6(-) (OS: HR=2.467, 95%CI 1.322-4.604, P=0.005; PFS: HR=2.248, 95%CI 1.275-3.965, P=0.005) and BCL2(S+) (PFS: HR=2.045, 95%CI 1.119-3.735, P=0.020) have worse prognosis. The co-expression of BCL6(-) and BCL2(S+) has significant influence on prognosis of DLBCL (P=0.005 and P<0.001) , in which BCL6(+)/non-BCL2(S+) (n=86) has the best prognosis[3-year-OS (71.6±4.9) %, 3-year-PFS (67.0±5.1) %], and BCL6(-)/BCL2(S+) (n=10) has the worst prognosis[3-year-OS (20.0±12.6) %, 3-year-PFS (10.0±9.5) %]; the co-expression of BCL6(-) and P53(+) has no significant influence on prognosis (P=0.061 and P=0.089) , however, those cases with BCL6(+)/P53(-) (n=98) often get better prognosis[3-year-OS (70.6±4.7) %, 3-year-PFS (64.6±4.9) %] than others; the co-expression of P53(+) and BCL2(S+) has significant influence on prognosis of DLBCL (P<0.001 and P<0.001) , and P53(+)/BCL2(S+) (n=5) has the worst prognosis (3-year-OS and 3-year-PFS are both 0) ; BCL2(S+) cases get shorter OS and PFS, regardless of the expression of BCL6 and P53. Conclusion: The expression and co-expression of BCL6 negative, P53 positive and BCL2(S+) have certain value in the prognostic evaluation of DLBCL, especially in the group of age≤60-year-old and IPI score 0-2.

[Clinicopathological features of intravascular peripheral T-cell lymphoma].

Objective: To summarize the clinical and pathological features of intravascular NK and T cell lymphoma for better understanding of such disease to reduce misdiagnosis and miss-diagnosis. Methods: Clinical and pathological features were analyzed retrospectively in one case of intravascular peripheral T-cell lymphoma, not otherwise specified (IVPTCL, NOS) , with literatures review. Results: The case presented in this study was a 66-year-old man. PET/CT scan showed multiple lymph nodes enlargement throughout the body. Normal lymph node structure could not be observed by tissue biopsy, while lymph follicles were partially disrupted. High-power light microscope revealed a large number of blood vessels with diffuse proliferation and dilation, where atypical lymphoid cell mass was restricted in the lumen and partially infiltrated the large blood vessel wall. These tumor cells were medium to large with moderate cytoplasm. The nucleus was irregular, single or multiple nucleoli could be seen, chromatin was condensed, some were empty and bright, and mitotic figures could be seen. Immunohistochemical staining showed that the neoplastic cells were positive for expression of CD3, CD43, CD8, GrB, TIA-1 and perforin. EBER in situ hybridization result was negative. Polymerase chain reaction test identified a clonal gene rearrangement of T-cell receptor γ. The patient was treated with CHOP in combination with chidamide, but died of infection and cardiopulmonary failure within 2 months. 56 cases of intravascular NK/T cell lymphoma with definite classification were collected from relevant literatures, including 47 cases with nasal type of extranodal NK/T cell lymphoma (27 were male and 20 were female) , 8 cases with anaplastic large cell lymphoma (3 males and 5 females) , and only one case with de nova IVPTCL, NOS in brain. We report the second case of IVPTCL,NOS, and notably originated from lymph node for the first time. Conclusions: Intravascular NK/T cell lymphoma is a highly aggressive disease with no effective treatment at present. Involvement of Lymph node has rarely been reported, and further studies on more cases are necessary.

A hyperspectral approach for recovering agent excretion biodistributions using whole-body fluorescence cryo-imaging.

Understanding the uptake and clearance kinetics of new drugs and contrast agents is an important aspect of drug development that typically involves a combination of imaging and analysis of harvested organs. Although these techniques are well-established and can be quantitative, they generally do not preserve high resolution biodistribution information. In this context, fluorescence whole-body cryo-imaging is a promising technique for recovering 3D drug/agent biodistributions at a high resolution throughout an entire study animal at specific time points. A common challenge associated with fluorescence imaging in tissue is that agent signal can be confounded by endogenous fluorescence signal which is often observed in the visible window. One method to address this issue is to acquire hyperspectral images and spectrally unmix agent signal from confounding autofluorescence signals using known spectral bases. Herein, we apply hyperspectral whole-body cryo-imaging and spectral unmixing to examine the distribution of multiple fluorescent agents in excretion organ regions.

Herbaceous Dominant the Changes of Normalized Difference Vegetation Index in the Transition Zone Between Desert and Typical Steppe in Inner Mongolia, China.

Asymmetric responses of aboveground net primary productivity (ANPP) to precipitation were identified as a signal to predict ecosystem state shifts at temperate grassland zones in Inner Mongolia, China. However, mechanism studies were still lacking. This study hypothesized that the enhanced growth and newly emerged herbaceous after increased precipitation resulted in the highest asymmetry at the transition zone between desert and typical steppe. We monitored the responses of the normalized difference vegetation index (NDVI) of different species to precipitation events using un-manned aerial vehicle technology to test this hypothesis. NDVI and species richness were measured twice at fixed points in July and August with a time interval of 15 days. Results showed that: (1) From July to August, NDVI in the transition zone increased significantly after precipitation (P < 0.05), but NDVI in both the desert and typical steppe showed a non-significant change (P > 0.05). (2) In the transition zone, NDVI increases from the shrub and herbaceous contributed to 37 and 63% increases of the site NDVI, respectively. (3) There was a significant difference in species richness between July and August in the transition zone (P < 0.05), mainly caused by the herbaceous (Chenopodiaceae, Composite, Convolvulaceae, Gramineae, Leguminosae, and Liliaceae), which either emerged from soil or tillers growth from surviving plants. This study demonstrated that herbaceous dominant the changes of NDVI in the transition zone, which provides a scientific basis for the mechanism studies of ANPP asymmetric response to precipitation and warrants long-term measurements.

[Quality-based soil fertility quality assessment of American ginseng main production areas in Shandong province].

The soil fertility quality is one of the most critical indicators of soil productivity. It directly affects the yield, quality and agricultural efficiency of Chinese medicinal materials. In order to establish the American ginseng planting soil fertility quality evaluation method based on the effective components of American ginseng, Wendeng district, Weihai city, Shandong province, the main producing area of American ginseng, was cited as a case for the study. Twenty-two 4-years American ginseng sampling sites are located at 7 towns. The samples of soil and plant root were collected in the autumn of 2017-2019. The saponin contents of American ginseng and 11 soil chemical properties were measured. The minimum data set(MDS) for assessment of the quality of soil fertility quality was established by correlation analysis and principal component analysis. The evaluation indexes were normalized by membership function. Soil quality index(SQI) that indicates soil comprehensive fertility quality level was calculated according to the critical value of membership function and weight value of each soil index in MDS. The results showed that the total saponin(Rg_1+Re+Rb_1) content of American ginseng in samples ranged from 1.76% to 7.94%. The yield of 8 plots in 2019 ranged from 3 818.7 kg·hm~(-2) to 8 996.4 kg·hm~(-2). MDS includes organic matter, alkaline nitrogen, exchangeable calcium, exchangeable magnesium, effective iron, effective copper, and effective zinc. Based on the mean of 4.825% of total saponin, threshold value of SQI for the region was determined to be 0.15, and 86.36% of soil samples in the county were above the threshold value. The methods and parameters are applicable to selection of high quality American ginseng planting sites and guiding rational fertilization. It also provides a reference for the evaluation of soil fertility quality of other medicinal plants.

[Effect of fragmented sleep on postoperative cognitive function and central neuroinflammation].

Objective: To investigate the effect of sleep fragmentation on perioperative neurocognitive disorders (PND) and central neuroinflammation by simulating sleep patterns of postoperative patients with sleep fragmentation in aged mice. Methods: Thirty-two elderly ICR mice were randomly divided into four groups (n=8): normal group (C), surgery group (S), fragmented sleep group (F), and surgery+fragmented sleep group (D). Fragmented sleep was conducted after internal fixation of tibia fractures, cognitive function was evaluated by novel object recognition (NOR) and fear conditioning (FC) test, and changes in expression of inflammatory cytokines in hippocampus were detected by ELISA. Results: NOR test: the recognition index (RI) of mice in group C, group S, group F and group D was 0.69±0.07, 0.48±0.07, 0.54±0.10 and 0.50±0.12, respectively. The RI of mice in group S, group F and group D was significantly lower than that in group C (t=4.885, 3.521 and 4.433, all P<0.01). There was no significant difference in RI between group S and group D (t=0.967 1, P>0.05). Contextual FC test: the freezing time of mice in group C, group S, group F and group D was(21.34±6.48), (13.83±4.26), (11.50±6.25) and (6.17±4.77) s, respectively. The freezing time of mice in group S, group F and group D was significantly lower than that in group C (t=2.722, 3.566, 5.496, P<0.05 or P<0.01). The freezing time of mice in group D was significantly lower than that in group S (t=2.774, P<0.05). Cue FC test: the freezing time of mice in group C, group S, group F and group D was (74.36±17.09), (43.91±9.71), (46.34±13.43) and (24.90±14.21) s, respectively. The freezing time of mice in group S, group F and group D was significantly lower than that in group C (t=4.393, 4.043 and 7.136, all P<0.01). The freezing time of mice in group D was significantly lower than that in group S (t=2.743, P<0.05). The levels of TNF-α, IL-6 and IL-1ß in hippocampus of mice in group S, F and D were significantly higher than those in group C, while the levels of TNF-α and IL-6 in hippocampus of mice in group D were significantly higher than those in group S, with statistically significant differences (P<0.05 or P<0.01). Conclusion: Postoperative fragmented sleep aggravates postoperative cognitive impairment and increases the hippocampal neuroinflammation in aged mice.

Negative magnetization, complex magnetic ordering and applications of Cr-doped Co2TiO4.

Polycrystalline Co2Ti1-xCrxO4 (0 ≤ x ≤ 0.2) inverse spinel ceramics have been synthesized via a sol-gel technique. The dc magnetization measurement in the field-cooled mode shows that negative magnetization could be observed until x reaches 0.2. The exchange constants are calculated using the ferrimagnetic Curie-Weiss fitting and the mean-field theory. This reveals that the strength of the inter sublattice magnetic interaction presents a non-monotonic trend with the increase in Cr content and reaches the minimum at x = 0.1, giving rise to the highest compensation temperature in the x = 0.1 sample. The applicability of the x = 0.1 sample is investigated in light of two prominent magnetic effects: (i) the stable magnetic switching effect indicates the potential applications in magnetic switching and data storage and (ii) the coexistence of normal and inverse magnetocaloric effects suggests a potential application in a constant temperature bath at 54 K.

Evaluating uptake of multiple fluorescent contrast agents in brain tumors simultaneously using whole animal multi-spectral cryo-imaging.

MRI images of gadolinium-based contrast agents (GBCA's) acquired before surgery are often registered to patients and used to guide surgical resection of intracranial tumors. Yet, the accuracy of these MR images in describing the surgical field degrades as surgery progresses; a well-recognized problem which has prompted efforts to develop new techniques that provide updated guidance information on residual tumor location. These efforts span a wide array of technologies, including image updating with deformation models, intraoperative MRI, and fluorescence guided surgery, among others. However, introduction of a straightforward technique that provides surgeons with a current view of GBCA distribution in real time remains an important goal. In this context, development of a fluorescent agent that recapitulates the kinetic behavior of GBCA's could provide familiar information directly in the surgical field in real time. To advance this strategy, we have begun identifying fluorescent contrast agents that show similar kinetic behavior to GBCA's. Using a novel hyperspectral whole body cryo-imaging system, we acquired high-resolution 3-D volumes of the distribution of multiple candidate fluorophores in whole heads bearing orthotopic brain tumors. Preliminary results reveal significant differences in the distribution of candidate optical agents, some of which show strong similarity to the GBCA uptake. Identification and eventual translation of a reliable GBCA-optical analog could improve and simplify surgical resection of brain tumors.

[Modified memory sub-test of Syndrom Kurz test in middle-aged and elderly Chinese].

Objective: To investigate the applicability of the modified memory sub-test of syndrom kurz test (SKT-M) in middle-aged and elderly Chinese. Methods: Between March 1, 2017, and October 31, 2017, at HwaMei Hospital, 132 patients receiving elective great saphenous vein high ligation and stripping operation and 96 their accompanying dependents, 55-75 years old, were randomly divided into the SKT-M group (n=121) and auditory verbal learning test -huashan version (AVLT-H) group (n=107) using random numeral method. The two groups underwent two corresponding neuropsychological tests respectively on the day before surgery and the second day after surgery. Results: There was no significant difference in the baseline characteristics and all the neuropsychological indices at the two time points between patients and dependents (P>0.05). As a consequence, the data of the patients and dependents were integrated to compare the applicability of SKT-M and AVLT-H. The "low-score" ratio of SKT-M immediate recall (2.4%) was lower than that of AVLT-H test (12.1%) (χ(2)=8.138, P<0.01). Besides, the "low-score" ratio of SKT-M delayed recall (5.7%) was also lower than that of AVLT-H test (20.5%) (χ(2)=11.167, P<0.01). The influence factors of SKT-M were less than that of AVLT-H test. However, the learning effect of SKT-M immediate recall was more significant, for its first testing sore (23.1±5.4) was significantly higher than the second one (21.9±5.1) (t=-3.971, P<0.001). Conclusion: The SKT-M has better applicability to 55-75 years old Chinese than AVLT-H test, but its learning effect should be noted.

A rare case of thoracic spinal intradural extramedullary enterogenous cyst with acute onset: case report and literature review.

The intraspinal enterogenous cyst, also called an neurenteric cyst, is a rare congenital disease. It was reported to be local to the C1 to L2 spinal segments, with the majority located in the cervicothoracic region. Most patients present with symptoms of progressive focal pain, myelopathic signs or radicular symptoms. We report a rare case of thoracic spinal intradural extramedullary enterogenous cyst with rapidly progressive weakness of both lower extremities. Additionally, we analysed the literature concerning the clinical features, diagnosis and prognosis of this disease.

[Experimental studies on the repair and restitution of cartilage by cartilage acellular extracellular matrix and adipose tissue-derived stem cells].

Objective: To investigate the effects of the repair and restitution of ear-shaped cartilage by adipose tissue-derived stem cells(ADSC) and cartilage acellular extracellular matrix. Methods: ADSC were extracted by digesting with collagenase type II from the adipose tissue from 32 patients with adiposity whose fats were drawn, and were cultured and subcultured in vitro. The natural biological scaffolds were prepared by acellular method using porcine ear cartilage, and then the second generation ADSC(5.0×10(7)/ml) were inoculated on the preformed natural bio-scaffold scaffold by culturing in vitro for 3 days to form a cell scaffold complex. 32 New Zealand white rabbits were randomly divided into the experimental groups, the control group A, the control group B and the control group C. All New Zealand white rabbits were modeled by ear cartilage defects. The cell scaffolds composite was implanted into the experimental group of the ear cartilage defects of rabbits, the ADSC were implanted into the control group A, the cartilage acellular extracellular matrix scaffold was implanted into the control group B and the control group C was modeled only by ear cartilage defects. After 16 weeks, the animals were sacrificed and the repair effect was observed by gross appearance and histological examinations including HE, Toluidine blue staining, Safranin O and typeⅡ collagen staining. Its were quantitatively analyzed by positive staining results of type Ⅱ collagen. Ear cartilage tissue elasticity was detected. SPSS 17.0 software was used to analyze the data. Results: The cartilage defects in the experimental group were repaired well by general shape observation and those in the control group was filled in with granulation tissue. There were significant differences between the experimental group and the control group in the wet weight(P<0.05). HE staining showed that cartilage cavities formed in articular cartilage defects, and only the fibrous tissue was filled with the ear cartilage defect in the control groups. In the repair area, Toluidine blue staining, Safranin O and type Ⅱ collagen staining were positive in the experimental group, and negative in the control groups. There was no significant difference between the experimental group and the normal ear cartilage in the ear cartilage elastic constant detection(P>0.05). Conclusions: The mechanics and histology of rabbit ear neonatal cartilage constructed by ADSC combined with cartilage acellular matrix are close to normal ear cartilage. Cartilage acellular matrix material combined with adipose-derived stem cells has good repair and reconstruction ability for ear cartilage defects, which possesses potential clinical application value.

[Prognostic evaluation of P53 and BCL2 proteins in MYC/BCL2 double expression DLBCL].

Objective: To investigate the strong expression (S+) of P53 and BCL2 proteins in MYC/BCL2 double-expression DLBCL (DEL) and whether they can be used for the prognostic evaluation and stratified diagnosis of DELs. Methods: Tissue microarray were made by filed FFPE blocks of 174 DLBCL cases. The translocation of MYC, BCL2 and BCL6 genes were detected by FISH, and the proteins were detected by IHC. Data of clinicopathologic features and follow up of patients were collected and OS (overall survival) and PFS (progression free survival) were analyzed by statistics. Results: Eight double-hit lymphomas (DHLs) were identified in all cases, and 45 DELs were selected from 166 remaining cases, which have no significant difference in OS and PFS compared with non-DEL cases (P=0.668 and P=0.790) . Of 42 DEL-cases with follow up data, 24 cases with P53+ or/and BCL2 (S+) are significantly shorter OS and PFS than others (P=0.003 and P=0.000) , in which the cases with P53+/BCL2 (S+) co-expression were the worst prognosis, and P53/BCL2 co-weaker positive DEL cases even have superior OS and PFS than those non-DELs. Although statistics showed that the cases of P53+ or/and BCL2 (S+) have a lower OS and PFS in total cases (P=0.063 and P=0.024) , it is not the case when the DEL-cases take out from total cases, that is the cases with P53+ or/and BCL2 (S+) are as similar OS and PFS as others in non-DEL group (P=0.590 and P=0.550) . Conclusion: The strong expression of P53 and BCL2 proteins can be used as indicators of stratified diagnosis and poor prognosis of DEL.

[Comparison of effects of two anesthesia methods on the first night sleep quality in middle-aged and elderly patients undergoing surgery for lower extremity varicose vein].

Objective: To investigate effects of two anesthesia methods on the first night sleep quality in middle-aged and elderly patients after surgery. Methods: This was a prospective study conducted from November 2017 to March 2018. Sixty patients, aged 50-70, undergoing elective surgery for unilateral lower extremity varicose vein at Ningbo No.2 Hospital, with American Society of Anesthesiologists (ASA) grade Ⅰ or Ⅱ, were enrolled and randomly allocated to two groups (n=30), general anesthesia group and spinal anesthesia group. On the first day before surgery, the patient's general data were collected and the Pittsburgh sleep quality index (PSQI) was used to assess the patient's sleep status in the past month. The postoperative mean arterial pressure (MAP), heart rate (HR) and pulse oxygen saturation (SpO(2)) in the ward were recorded with a multi-function monitor on first night after surgery. The total sleep time and arousal time were obtained by bispectral index (BIS) monitoring from 20: 00 (the first day) to 6: 00 (the second day). Athens Insomnia Scale (AIS) was recorded at 18: 00 at the second day after surgery. Results: There was no significant difference in general data and PSQI scale scores between the two groups of patients (all P>0.05). And there were no significant differences in MAP, HR, SpO(2) and BIS every 2 hours between the two groups from 20: 00 (the first day) to 6: 00 (the second day)(all P>0.05). Compared with the general anesthesia group, the first night of total sleep time in the spinal anesthesia group was significantly shorter[(357.2±83.4)min vs (275.1±64.8)min, t=-9.635, P<0.05], while the rate of wakefulness, total sleep time, overall sleep quality, daytime mood and daytime physical function were significantly higher[(25.9%, 22.2%, 25.9%, 18.5%18.5%) vs (51.7%, 51.7%, 55.2%, 48.3%44.8%), χ(2)=3.901, 5.192, 4.941, 5.523 and 4.437, all P<0.05], and the cases of postoperative urinary retention and lower limb discomfort were significantly higher[(8 and 6) vs (1 and 0), all P<0.05]. Conclusion: Both anesthesia methods can be safely and effectively applied to middle-aged and elderly patients with lower extremity varicose veins surgery, but patients with general anesthesia show fewer adverse reactions on the first night after surgery and have better sleep quality.

[A clinic analysis of thoracolumbar vertebral fracture cascade].

Objective: To observe the clinical characteristics of thoracolumbar vertebral fracture cascade, analyze the relationship between the baseline fractures and the subsequent fractures and compare the distribution differences of subsequent fractures following vertebral augmentation or non-operation. Methods: From July 2012 to August 2016, 1 363 patients admitted to the First Affiliated Hospital of Soochow University with vertebral augmentation for the treatment of vertebral fractures were retrospectively analyzed.There were 190 cases of vertebral fracture cascade, 160 females and 30 males, with an average age of (74±9) years.The location and sequence of all vertebral fractures were recorded.The relationships between the baseline and the subsequent fractures were analyzed.According to different treatment on the baseline vertebral fractures, 190 cases were divided into vertebral augmentation group and non-operation group.The distribution differences of the subsequent fractures following vertebral augmentation and non-operation were compared with chi-square test. Results: Vertebral fracture cascade mainly located in the thoracolumbar spine T(11)-L(2) with an incidence of 52.0%.According to the direction of fracture development, the fracture cascade could be divided into up, down, centrifugation and concentration, and the incidence was 39.8%, 39.2%, 8.4% and 12.6%, respectively.The closer the vertebral body to the baseline fractures, the subsequent fractures incidence was higher.For distance with zero, one, two, three and four vertebrae, the incidence of subsequent vertebral fractures was 36.5%, 26.2%, 15.2%, 11.5% and 3.7%, respectively.A linear relationship was found between the subsequent fractures and the baseline fractures with a correlation coefficient of 0.90.The distribution difference of subsequent fractures between vertebral augmentation and non-operation group was not significant (χ(2)=17.16, P>0.05). Conclusions: The main directions of vertebral fracture cascade is up or down spiral development.The closer the vertebral body to the baseline fractures, the subsequent fractures incidence is higher.Vertebral augmentation doesn't affect the distribution of subsequent fractures.