Correlation study between bone metabolic markers, bone mineral density, and sarcopenia.

OBJECTIVE: To investigate the correlation between bone metabolism markers, bone mineral density (BMD), and sarcopenia. METHODS: A total of 331 consecutive patients aged ≥ 60 years who were hospitalized between November 2020 and December 2021 were enrolled. Participants were divided into sarcopenia and non-sarcopenia groups according to the Asian Working Group on Sarcopenia criteria (AWGS, 2019). The clinical data, bone metabolism markers (ß-CTX, N-MID, and TP1NP), and BMD were compared between the two groups. RESULTS: Age, ß-CTX, and N-MID of the sarcopenia group were higher than those of the non-sarcopenia group (P < 0.05), but the BMD T values were lower than those of the non-sarcopenia group (P < 0.05). Binary logistic regression analysis showed that increased femoral neck BMD (FNBMD) was a protective factor for sarcopenia, while increased ß-CTX was a risk factor. Pearson/Spearman correlation analysis showed that the diagnostic indices of sarcopenia were positively correlated with FNBMD and negatively correlated with ß-CTX and N-MID. Multiple linear regression analysis revealed that BMI and FNBMD significantly positively affected muscle strength and appendicular skeletal muscle mass (ASM). The FNBMD significantly positively affected physical performance, while ß-CTX significantly negatively affected muscle strength, ASM, and physical performance. CONCLUSION: Increased FNBMD may be a protective factor against sarcopenia, and increased ß-CTX may be a risk factor. The FNBMD significantly positively affected the diagnostic indices of sarcopenia, while ß-CTX significantly negatively affected them. BMD and bone metabolism marker levels may be considered in early screening for sarcopenia.

[A study on the distribution characteristics of peripheral retinal defocus in children and adolescents].

Objective: To explore the distribution characteristics of peripheral retinal defocus in children and adolescents. Methods: This cross-sectional study included 500 individuals aged 3 to 18 years, who visited the People's Hospital of Lincang, the First Affiliated Hospital of Dali University and Dali Ophthalmology Hospital between January and December 2021. Data of the right eye of each participant was analyzed. There were 226 males (45.20%) and 274 females (54.80%), with an average age of (10.79±3.79) years. All participants underwent post-cycloplegic refraction, optical biometry, and intraocular pressure measurement to obtain spherical equivalent, average corneal curvature, axial length, and intraocular pressure. Multispectral refraction topography was performed to obtain topographic maps and values at various field angles and orientations of peripheral retinal defocus. Based on multispectral refraction topography, peripheral retinal defocus values were categorized as crater type, hemilateral upturn type, saddle type, and relatively flat type. The distribution of different refractive states was analyzed. Results: The spherical equivalent of the 500 participants was(-1.51±2.61) D, axial length was (24.10±1.28) mm, and average corneal curvature was (43.20±1.22) D. Among the 500 eyes, 382 exhibited hyperopic peripheral retinal defocus values, with 316 eyes (82.72%) being myopic. Myopic peripheral retinal defocus values were observed in 118 eyes, with 15 eyes (12.72%) being myopic. Among different types of peripheral retinal defocus values, 112 eyes (22.4%) exhibited a crater type, 153 eyes (30.6%) exhibited a hemilateral upturn type, 107 eyes (21.4%) exhibited a saddle type, and 128 eyes (25.6%) exhibited a flat type. The proportion of myopia was 82.14% (92 eyes), 69.28% (106 eyes), 60.75% (65 eyes), and 3.90% (5 eyes), respectively. The peripheral retinal defocus values at 15°, 30°, and 45° were (0.01±0.08) D, (0.06±0.21) D, and (0.20±0.37) D, respectively. The peripheral retinal defocus values at temporal, inferior, nasal, and superior locations were (0.58±0.69) D, (0.52±0.63) D, (0.21±0.64) D, and (-0.26±0.67) D, respectively. Notably, the superior primarily manifested as myopic, while the others were predominantly hyperopic. Conclusions: Approximately three-fourths of children and adolescents exhibit hyperopic peripheral retinal defocus values, with a higher prevalence of myopia in this subgroup. The hyperopia peripheral retinal defocus value increases with the distance from the retina to the macula. The peripheral retinal defocus values between superior and inferior, nasal and temporal locations are asymmetrical, with the temporal hyperopic peripheral retinal defocus value being most prominent and the superior myopic peripheral retinal defocus value being most evident.

[Advances in the epidemiology of chronic thromboembolic pulmonary hypertension].

The early symptoms of chronic thromboembolic pulmonary hypertension (CTEPH) are not specific, and there is a high rate of misdiagnosis, missed diagnosis, and lack of awareness among clinicians. Understanding the current epidemiological characteristics of CTEPH is helpful to raise the understanding level of Chinese clinicians on CTEPH and improve the current status of prevention and treatment. However, epidemiological information and relevant reviews on CTEPH are currently lacking in China. In this review, we combined the published epidemiological literature on CTEPH in the real world, summarized the research overview, prevalence, incidence, survival rate and risk factors of CTEPH, and provided an outlook for the development of multicenter and high-quality CTEPH epidemiological research in China.

Mitochondrial quality control in cartilage damage and osteoarthritis: new insights and potential therapeutic targets.

Osteoarthritis (OA) is a multifactorial arthritic disease of weight-bearing joints concomitant with chronic and intolerable pain, loss of locomotion and impaired quality of life in the elderly population. Although the prevalence of OA increases with age, its specific mechanisms have not been elucidated and effective therapeutic disease-modifying drugs have not been developed. As essential organelles in chondrocytes, mitochondria supply energy and play vital roles in cellular metabolism, proliferation and apoptosis. Mitochondrial quality control (MQC) is the key mechanism to coordinate various mitochondrial biofunctions, primarily through mitochondrial biogenesis, dynamics, autophagy and the newly discovered mitocytosis. An increasing number of studies have revealed that a loss of MQC homeostasis contributes to the cartilage damage during the occurrence and development of OA. Several master MQC-associated signaling pathways and regulators exert chondroprotective roles in OA, while cartilage damage-related molecular mechanisms have been partially identified. In this review, we summarized known mechanisms mediated by dysregulated MQC in the pathogenesis of OA and latent bioactive ingredients and drugs for the prevention and treatment of OA through the maintenance of MQC.

[The correlation between plasma heat shock proteins 90α levels and white matter hyperintensity in patients with cerebral small vessel disease].

Objective: To investigate the relationship between plasma heat shock proteins 90α(Hsp90α) levels and the white matter hyperintensity(WMH) in patients with cerebral small vessel disease(SVD). Methods: Patients admitted to the Department of Neurology, the First Affiliated Hospital of Zhengzhou University from March to August 2021 and diagnosed with WMH by magnetic resonance examination (MRI) were selected as the case group, matched with physical examination patients who visited the Department of Medical Examination during the same period and showed no WMH on MRI and no history of neurological diseases as the control group, and the level of plasma Hsp90α was quantitatively detected by enzyme-linked immunosorbent assay. Mann-Whitney U test was used to compare whether there was a difference in plasma Hsp90α levels between the control group and the case group.Multivariate logistic regression analysis was used to explore the related factors of WMH in patients with SVD. Results: Of the 183 subjects, the control group (n=73) consisted of 28 males and 45 females, aged (54±10) years, while the case group (n=110) consisted of 71 males and 39 females, aged (64±10) years old. Plasma Hsp90α level was higher in the case group than that of the control group [53.33(35.33, 70.09) ng/ml vs 35.02(18.51, 54.95) ng/ml, P<0.001]. After adjusting for confounding factors by multivariate analysis, the results showed that plasma Hsp90α levels greater than 58.34 ng/ml was associated with WMH (P=0.002, OR=5.931, 95%CI:1.955-17.995). Conclusion: Higher level of plasma Hsp90α is associated with WMH in patients with SVD.

[Recommendations for the medication of rheumatic diseases in periconceptional period].

Rheumatic diseases are a kind of chronic inflammatory diseases mainly involving joints and surrounding tissues. Most patients with rheumatic diseases need long-term treatment, which is difficult to be avoided during pregnancy. Treatment efficacy, as well as maternal and fetal safety should be taken into account in the medical decision. Based on the domestic and foreign guidelines, consensus, diagnosis and treatment experience, Chinese Rheumatology Association developed the standardization of medication use in patients with rheumatic diseases preparing and during pregnancy, aiming on the application and precautions of commonly used medicines for rheumatic diseases in preparing pregnancy, pregnancy and lactation.

[Rational use of antibiotics in severe acute pancreatitis:controversy and progress].

Infection of pancreatic necrosis is the most frequent cause of late mortality in severe acute pancreatitis(SAP). Most clinical guidelines of acute pancreatitis recommended that prophylactic antibiotics should be avoided. Prophylactic antibiotics can not reduce the pancreatic infection rate or mortality in patients with SAP ornecrotizing pancreatitis. Definitive infection is the only indication for rational use of antibiotics in SAP patients. Broad-spectrum antibiotics for treatment should cover enteric bacteria, and the bacteriology and antibiotic pharmacokinetics of SAP should be considered when selecting antibiotics.

[The role of intestinal failure in the pathogenesis of severe acute pancreatitis].

Severe acute pancreatitis can induce systemic and local complications, with infectious pancreatic necrosis and sepsis leading to the second death peak. Enterogenous infection caused by intestinal failure is considered to be an important mechanism of secondary infection of pancreatic or peripancreatic necrosis. Therefore, the prevention and treatment of intestinal failure is the key point in the treatment of severe acute pancreatitis and has an important influence on the course and prognosis of the disease. Individualized treatment should be selected according to the advantages of treatment centers and the characteristics of patients.

[Safety of laparoscopic cholecystectomy for gallbladder cancer: a meta-analysis].

Objective: To evaluate the effect and safety of laparoscopic cholecystectomy (LC) on the treatment of patients with gallbladder cancer (GBC), compared with patients undergoing open cholecystectomy (OC). Methods: PubMed, EMBASE, Web of Science, CNKI, CQVIP and WANFANG DATA and the Cochrane Library were searched for all Chinese and English literatures of randomized or non-randomized concurrent controlled trials of OC and LC treatment of GBC from the database establishment to March 2020. Two reviewers selected the studies according to inclusion and exclusion criteria, extracted the data, and then a meta-analysis was subsequently performed by the RevMan 5.3 software provided by the Cochrane Library. Results: A total of 15 studies (1 074 patients) including 14 retrospective studies and 1 prospective study met the inclusion criteria. The meta-analysis showed that compared with OC, LC has significant short-term efficacy in the treatment of patients with gallbladder cancer, including shorter operation time (mean difference (MD) =-18.78, 95% confidence interval (CI) : -36.68-0.88; P=0.04), less intraoperative blood loss (MD=-166.57, 95%CI: -248.51--84.63; P<0.000 1), shorter post-operative hospital stays (MD=-5.00, 95%CI: -6.43--3.57; P<0.000 1), less complication rate (OR=0.47, 95%CI: 0.28-0.79; P=0.004), but there was no significant difference on the aspects of recurrence rate (OR=0.48, 95%CI: 0.21-1.11; P=0.09), 5-year overall survival (HR=0.93, 95%CI: 0.54-1.61, I2=33.5%, P=0.198) and long-term survival. Conclusion: Whether it is radical cholecystectomy (RC) or simple cholecystectomy (SC), the short-term efficacy of LC is more significant than that of OC, and the long-term survival rate has no significant statistical difference. Limited by the quality of literature and experiments, the above conclusions still need to be supported by higher quality research.

[Clinicopathological analysis of 16 cases of pyrrolizidine alkaloids-associated hepatic sinusoidal obstruction syndrome].

Objective: To observe the histopathological manifestations of liver biopsy in patients with hepatic sinusoidal obstruction syndrome (HSOS) induced by pyrrolizidine alkaloid (PA). Methods: Patients diagnosed with PA-HSOS from 2012 to 2017 were selected, and the general conditions, liver function indexes, medication history, liver biopsy time, histopathological slides of liver biopsy, and follow-up data of clinical prognosis after 6 months of onset were collected. Clinical staging with clinical data was used to observe the histopathological manifestations of patients at different clinical stages. Wilcoxon rank-sum test, unpaired t-test and univariate linear regression analysis were used for data analysis. Results: A total of 16 cases were collected. Alanine transaminase and aspartate transaminase was 59.25 U/L and 25.50 U/L, 108 U/L and 45 U/L, respectively, after 6 months of onset and follow-up, and the differences were statistically significant. Moreover, total bile acids and albumin was 35 µmol/L and 36.15 µmol/L, and 32.45 g/L and 31 g/L, respectively, and the differences were not statistically significant. PA-HSOS pathological development process was divided into early, middle and late stages. In the early stage, the central lobular sinusoidal endothelium integrity was impaired and the entry of erythrocytes had interspersed thin reticular fibers and perisinusoidal space. In the middle stage (hemorrhagic zone), erythrocytes, reticular fibers and collagen fibers were lysed, densely collapsed and deposited. The cavity of the bloodstream was hyperemic and dilated, and the cavity was covered with sinus endothelial cells. The hepatic plate regenerated around the hemorrhagic zone and some of the hepatic sinuses were decompensated. In the late stage, deposited collagen in the hemorrhagic zone had formed a large fibrous scar, and most of the dilated cavity in the bloodstream was covered with vascular endothelium. The marginal zone hepatic cells were regenerated in two rows and gradually inserted into the fibrous septum. Different hepatic lobular lesions obtained from the same patients liver biopsy tissues were changed at different stages. Hepatic lobule injury proportion with severe internal bleeding in liver biopsy tissue had no relation with the prognosis of patients. Conclusion: In the early stage of PA-HSOS, erythrocytes in the central zone of lobules enter the perisinusoidal space through the damaged sinus endothelium, which is manifested as hepatic plate hemorrhagic necrosis. In the middle and late stage, liver plate regeneration and vascular remodeling occurred, so most of the patients' clinical course was self-limited. Pathological staging and liver biopsy time have an apparent correlation, but the prognosis of patients cannot be judged based on the extent of hemorrhage and injury of biopsy samples.

[Analysis of treatment modalities and prognosis of patients with gallbladder cancer in China from 2010 to 2017].

Objective: To evaluate the clinical characteristics and prognosis of gallbladder cancer (GBC) patients in China. Methods: This retrospective multicenter cohort study enrolled 3 528 consecutive GBC patients diagnosed between January 2010 to December 2017 in 15 hospitals from 10 provinces. There were 1 345 (38.12%) males and 2 183 (61.88%) females.The age of diagnosis was (63.7±10.8) years old (range: 26 to 99 years old) .There were 213 patients (6.04%) in stage 0 to Ⅰ, whereas 1 059 (30.02%) in stage Ⅱ to Ⅲ, 1 874 (53.12%) in stage Ⅳ, and 382 (10.83%) unavailable. Surgery was performed on 2 255 patients (63.92%) . Three hundred and thirty-six patients received chemotherapy or radiotherapy (9.52%; of which 172 were palliative); 1 101 (31.21%) received only supportive treatment.The patient source, treatment and surgery, pathology, concomitant gallstone, and prognosis were analyzed. Results: Among the 3 528 GBC patients, 959 (27.18%) were from East China, 603 (17.09%) from East-North China, 1 533 (43.45%) from Central China, and 433(12.27%) from West China. Among the 1 578 resectable tumor, 665 (42.14%) underwent radical surgery, 913 (57.86%) underwent surgery that failed to follow the guidelines.Eight hundred and ninety-one (56.46%) patients were diagnosed before surgery, 254 (16.10%) during surgery, and 381 (24.14%) after surgery (time point of diagnosis couldn't be determined in 52 patients) .Among the 1 578 patients with resectable tumor, 759 (48.10%) had concomitant gallstone.Among the 665 patients underwent radical surgery, 69 (10.4%) showed positive resection margin, 510 (76.7%) showed negative resection margin, and 86 (12.9%) unreported margin status.The 5-year overall survival rate (5yOS) for the 3 528-patient cohort was 23.0%.The 5yOS for patients with resectable tumor was 39.6%, for patients with stage ⅣB tumor without surgery was 5.4%, and for patients with stage ⅣB tumor underwent palliative surgery was 4.7%. Conclusions: More than half GBC patients in China are diagnosed in stage Ⅳ.Curative intent surgery is valuable in improving prognosis of resectable GBC.The treatment of GBC needs further standardization.Effective comprehensive treatment for GBC is in urgent need.

Comparative studies of mucosal humoral and cellular immune responses to 2009 pandemic H1N1 influenza virus in mice.

The nasal-associated lymphoid tissues (NALTs), embedded in the submucosa of murine upper respiratory tract, represents an important site of induction for local mucosal immune responses to airborne pathogens and intranasal vaccines. Here, we systematically investigated the mucosal humoral and cellular immune responses of NALTs in mice infected with A/Beijing/501/2009 (BJ501) and A/Puerto Rico/8/1934 (PR8) viruses. Compared with PR8 infection, BJ501 induced a more rapid increase of virus-specific IgA and IgG antibodies in the nasal lavage fluid and a higher ratio of IgG1/IgG2a, indicating a stronger Th2 response to BJ501 in mucosal immunity. In addition, using virus-specific enzyme-linked immunospot assay (ELISpot assay), we observed higher and earlier responses of virus-specific IgA and IgG antibody-secreting cells (ASCs) and IFN-γ and IL-4 cytokine-secreting cells (CSCs) in NALTs of mice intranasally infected with BJ501 virus. In particular, the frequency of BJ501-specific IFN-γ-CSCs significantly correlated with the kinetics of BJ501 virus load in NALTs, suggesting an important role of IFN-γ-CSCs-associated mucosal cellular immune responses in BJ501 virus clearance. Collectively, BJ501 induced a more comprehensive and rapid mucosal immune responses in NALTs of mice, providing further understanding of the immune responses elicited by 2009 pandemic H1N1 virus in upper respiratory tract. Keywords: nasal-associated lymphoid tissues (NALTs); influenza virus; mucosal immune response; Th1/Th2 response.

Unique genetic profiles from cerebrospinal fluid cell-free DNA in leptomeningeal metastases of EGFR-mutant non-small-cell lung cancer: a new medium of liquid biopsy.

Background: Leptomeningeal metastases (LM) are more frequent in non-small-cell lung cancer (NSCLC) with epidermal growth factor receptor (EGFR) mutations. Due to limited access to leptomeningeal lesions, the purpose of this study was to explore the potential role of cerebrospinal fluid (CSF) as a source of liquid biopsy in patients with LM. Patients and methods: Primary tumor, CSF, and plasma in NSCLC with LM were tested by next-generation sequencing. In total, 45 patients with suspected LM underwent lumbar puncture, and those with EGFR mutations diagnosed with LM were enrolled. Results: A total of 28 patients were enrolled in this cohort; CSF and plasma were available in 26 patients, respectively. Driver genes were detected in 100% (26/26), 84.6% (22/26), and 73.1% (19/26) of samples comprising CSF cell-free DNA (cfDNA), CSF precipitates, and plasma, respectively; 92.3% (24/26) of patients had much higher allele fractions in CSF cfDNA than the other two media. Unique genetic profiles were captured in CSF cfDNA compared with those in plasma and primary tissue. Multiple copy number variations (CNVs) were mainly identified in CSF cfDNA, and MET copy number gain identified in 47.8% (11/23) of patients was the most frequent one, while other CNVs included ERBB2, KRAS, ALK, and MYC. Moreover, loss of heterozygosity (LOH) of TP53 was identified in 73.1% (19/26) CSF cfDNA, which was much higher than that in plasma (2/26, 7.7%; P < 0.001). There was a trend towards a higher frequency of concomitant resistance mutations in patients with TP53 LOH than those without (70.6% versus 33.3%; P = 0.162). EGFR T790M was identified in CSF cfDNA of 30.4% (7/23) of patients who experienced TKI progression. Conclusion: CSF cfDNA could reveal the unique genetic profiles of LM and should be considered as the most representative liquid biopsy medium for LM in EGFR-mutant NSCLC.

Bone morphogenetic proteins for articular cartilage regeneration.

Degeneration of articular cartilage (AC) tissue is the most common cause of osteoarthritis (OA) and rheumatoid arthritis. Bone morphogenetic proteins (BMPs) play important roles in bone and cartilage formation. This article reviews the experimental and clinical applications of BMPs in cartilage regeneration. Experimental evidence indicates that BMPs play an important role in protection against cartilage damage caused by inflammation or trauma, by binding to different receptor combinations and, consequently, activating different intracellular signaling pathways. Loss of function of BMP-related receptors contributes to the decreased intrinsic repair capacity of damaged cartilage and, thus, the multifunctional effects of BMPs make them attractive tools for the treatment of cartilage damage in patients with degenerative diseases. However, the development of BMP therapy as a treatment modality for cartilage regeneration has been hampered by certain factors, such as the eligibility of participants in clinical trials, financial support, drug delivery carrier safety, availabilities of effective scaffolds, appropriate selection of optimal dose and timing of administration, and side effects. Further research is needed to overcome these issues for future routine clinical applications. Research and development leading to the successful application of BMPs can initiate a new era in the treatment of cartilage degenerative diseases like OA.

[The effects of microRNA-7 on proliferation and invasion of hepatocellular carcinoma HepG2 cells].

Objective: To investigate the effects of overexpression of microRNA-7 (miR-7) on the proliferation and invasion of HepG2 cells and the underlying mechanism in vitro. Methods: The relative expression levels of miR-7 and Raf1 in hepatocellular carcinoma (HCC) tissues and adjacent normal tissues (ANT) were detected by quantitative real time-PCR (qRT-PCR). The relationship between the expression of miR-7 and the characteristics of HCC patients was analyzed. Cells were divided into blank control group, negative control (NC) group and miR-7 mimics transfected group, miR-7 mimics and NC were transfected into HepG2 cells by Lipofectamine™2000. The relative expression of miR-7 was detected by qRT-PCR. The proliferation ability of HepG2 cells was detected by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay. The invasion of HepG2 cells was detected by Transwell assay. The target genes of miR-7 were predicted by TargetScan and the binding effect of miR-7 on the 3'UTR of Raf1 was verified by dual luciferase reporter assay.The expressions of Raf1 protein in hepatocellular carcinoma tissues, normal tissues and miR-7 mimics transfected HepG2 cells was detected by Western blot. The correlation of the levels of miR-7 and Raf1 mRNA was determined by Pearson correlation analysis. Results: The relative expression level of miR-7 in HCC was 0.49±0.02, significantly lower than in ANT (1.21±0.05, P<0.01). The level of miR-7 was significantly correlated the tumor volume, metastasis and prognosis of HCC patients (P<0.05). The relative expression level of miR-7 in miR-7 mimics transfected HepG2 group was 12.67±0.40, significantly higher than that in blank group (P<0.01). Compared with the blank group, the A value and invasion ability of miR-7 mimics transfected group were significantly down-regulated at 48 hours and 72 hours after transfection (P<0.01). Compared with miR-7 NC group, the luciferase activity of wild-type Raf1 reporter gene in miR-7 mimics transfected group was significantly reduced (P<0.01). The relative expression of Raf1 protein in HCC was 3.15±0.10, significant higher than in ANT (0.53±0.03, P<0.01). The relative expression of Raf1 protein in miR-7 mimics transfected group was 0.24±0.01, significantly lower than in miR-7 NC group (0.98±0.02, P<0.01). Furthermore, an negative correlation was observed between the levels of miR-7 and Raf1 in HCC tissues (P<0.05). Conclusions: The expression of miR-7 in HCC is significantly decreased and inversely correlated with poor survival of HCC patients. Overexpression of miR-7 can inhibit the proliferation and invasion ability of hepatocellular carcinoma cells HepG2 by downregulating Raf1 in vitro.

[Establishment and optimization of hepatocyte steatosis model].

Objective: To establish a new model of hepatic steatosis cells by optimizing the original ethanol or high fat, the present study proposed an in vitro hepatocyte steatosis model for the study of fatty liver. Methods: Oil red O staining was used to observe the effects of fetal bovine serum, oleic acid and ethanol on lipid accumulation in human liver cell line L02 in a concentration- and time-dependent manner. RT-PCR was used to detect the mRNA expression levels of PPAR-γ and AP-2, and the suitable conditions for the establishment of hepatocyte steatosis model were screened out. A t-test was used for comparison between the two groups, and one-way Analysis of Variance (ANOVA) was used in more than three groups. Results: Oil red O staining showed the number of reddish-orange lipid droplets in L02 cells gradually increased with the increase of fetal bovine serum, oleic acid and ethanol in a concentration - and time-dependent manner. Compared with 0.00% oleic acid and 2% ethanol, the count value of red particle was 100.00% ± 17.63% at the beginning and after 24 h, 0.003% oleic acid and 2% ethanol jointly acted in L02 cells. After incubation for 48 hours with 2% ethanol and serum-free DMEM medium, the accumulation of lipid droplets was the highest with a count value of 802.38%+71.06%(t = 42.36, P < 0.001). RT-PCR analysis showed the lipid accumulation induced by this method was positively correlated with the mRNA expression of PPAR-γ and AP-2. Conclusion: L02 cells were successfully exposed to high fat and ethanol, and the hepatocyte steatosis model was established and optimized, suggesting that the occurrence of hepatic cell steatosis was related to the up-regulation of PPAR-γ and AP-2.

A clip domain serine protease involved in moulting in the silkworm, Bombyx mori: cloning, characterization, expression patterns and functional analysis.

Clip domain serine proteases (CLIPs), characterized by one or more conserved clip domains, are essential components of extracellular signalling cascades in various biological processes, especially in innate immunity and the embryonic development of insects. Additionally, CLIPs may have additional non-immune functions in insect development. In the present study, the clip domain serine protease gene Bombyx mori serine protease 95 (BmSP95), which encodes a 527-residue protein, was cloned from the integument of B. mori. Bioinformatics analysis indicated that BmSP95 is a typical CLIP of the subfamily D and possesses a clip domain at the N terminus, a trypsin-like serine protease (tryp_spc) domain at the C terminus and a conserved proline-rich motif between these two domains. At the transcriptional level, BmSP95 is expressed in the integument during moulting and metamorphosis, and the expression pattern is consistent with the fluctuating 20-hydroxyecdysone (20E) titre in B. mori. At the translational level, BmSP95 protein is synthesized in the epidermal cells, secreted as a zymogen and activated in the moulting fluid. Immunofluorescence revealed that BmSP95 is distributed into the old endocuticle in the moulting stage. The expression of BmSP95 was upregulated by 20E. Moreover, expression of BmSP95 was downregulated by pathogen infection. RNA interference-mediated silencing of BmSP95 led to delayed moulting from pupa to moth. These results suggest that BmSP95 is involved in integument remodelling during moulting and metamorphosis.

Neuromolecular imaging, a nanobiotechnology for Parkinson's disease: advancing pharmacotherapy for personalized medicine.

Evaluating each patient and animal as its own control achieves personalized medicine, which honors the hippocratic philosophy, explaining that "it is far more important to know what person has the disease than what disease the person has." Similarly, individualizing molecular signaling directly from the patient's brain in real time is essential for providing prompt, patient-based treatment as dictated by the point of care. Fortunately, nanotechnology effectively treats many neurodegenerative diseases. In particular, the new medicinal frontier for the discovery of therapy for Parkinson's disease is nanotechnology and nanobiotechnology. Indeed, the unique nanotechnology of neuromolecular imaging combined with the series of nanobiosensors enables continuous videotracking of molecular neurotransmitters in both the normal physiologic and disease states with long-term electrochemical operational stability. This nanobiotechnology is able to track a signal in real time with excellent temporal and spatial resolution directly from each patient's brain to a computer as subjects are behaving during movement, normal and/or dysfunctional including prion-like Parkinson's behavioral biometrics. Moreover, the molecular signaling performed by these nanobiosensors live streams directly online and originates from precise neuroanatomic brain sites such as, in this case, the dorsal striatum in basal ganglia. Thus, the nanobiotechnology studies discussed herein imaged neuromolecules with and without L-3,4-dihydroxyphenylalanine (L-DOPA) in dorsal striatal basal ganglia neurons. Parkinsonian and non-Parkinsonian animals were video-tracked, and images were readily seen on a laptop via a potentiostat using a semiderivative electrical circuit. Administered L-DOPA doses were 50 and 100 mg/kg intraperitoneally (ip); the same experimental paradigm was used to image and then contrast data. Results showed that the baseline release of biogenic amine molecules was significantly above detection limits in non-Parkinsonian animals. After administration of L-DOPA, biogenic amines significantly increased in these non-Parkinson's animals. Nevertheless, it is intriguing to see that L-DOPA could not enable synaptic dopamine release in Parkinson's animals, thereby demonstrating that biogenic amines are biomarkers for Parkinson's disease. Biomarkers are biochemical, genetic, or molecular measures of biological reactions. Importantly, there were other significant biomarkers present in Parkinsonian animals and absent in non-Parkinsonian animals; these were peptide neurotransmitters that include dynorphin and somatostatin in the brain with detection limits of 40 nM for dynorphin and 37 nM for somatostatin (see Table 1). Furthermore, L-DOPA significantly increased these peptide biomarkers, dynorphin and somatostatin, in Parkinson's animals. Targeting biomarkers enables new diagnostic devices and treatments for Parkinson's disease through nanotechnology and nanobiotechnology.