Objective:To establish and validate a method for the determination of particle size and size distribution of gefitinib. Methods:A Malvern Mastersizer 3000 laser size analyzer and a Hydro LV wet autosampler were used,a light scattering method for the determination of particle size and size distribution was adopted to analyze the particle size of gefitinib,and then the methodological study was carried out. The pump speed was 2 000 r·min -1 ,the obscuration was 8% -20% ,the measurement time of background and sample was 10 s,the sample RI was 1. 500,and the sample absorbance was 0. 00. Results:The D10 of 3 batches of gefitinib was less than 7 μm,the D50 was less than 15 μm and the D90 was less than 25 μm,which all met the preparation requirements of final products. Conclusion:The method is accurate,simple and repeatable,and suitable for the particle size analysis of gefitinib.

OBJECTIVELow-intensity pulsed ultrasound (LIPUS) has been reported to enhance proliferation and to alter protein production in various kinds of cells. In the present study, we measured the neurites length after LIPUS treatment to define the effectiveness of LIPUS stimulation on neurons, and then we examined the acticity of GSK-3beta to study the intracellular mechanism of neurite's outgrowth.

METHODSLIPUS was applied to cultured primary rat cortical neurons for 5 minutes every day with spatial- and temporal average intensities (SATA) of 10 mW/cm(2), a pulse width of 200 microseconds, a repetition rate of 1.5 KHz, and an operation frequency of 1 MHz. Neurons were photographed on the third day after LIPUS treatment and harvested at third, seventh, and tenth days for immunoblot and semi-quantitative RT-PCR analysis.

RESULTSMorphology change showed that neurite extension was enhanced by LIPUS. There was also a remarkable decrease of proteins, including p-Akt, p-GSK-3beta, and p-CRMP-2, observed on the seventh and tenth days, and of GSK-3beta mRNA expression, observed on the seventh day, in neurons treated with LIPUS.

CONCLUSIONLIPUS can enhance elongation of neurites and it is possible through the decreased expression of GSK-3beta.

Animals ; Base Sequence ; Cells, Cultured ; DNA Primers ; Glycogen Synthase Kinase 3 ; antagonists & inhibitors ; Glycogen Synthase Kinase 3 beta ; Neurites ; Protein Kinase Inhibitors ; pharmacology ; Rats ; Reverse Transcriptase Polymerase Chain Reaction ; Ultrasonics

OBJECTIVETo investigate the techniques and clinical applications of multislice helical computed tomography (CT) colonography in colonic lesions.

METHODSFifty-nine patients with malignant lesions of colon underwent volume scanning using multislice helical CT. Four types of reconstruction including CT virtual colonoscopy (CTVC), shaded surface display (SSD), Raysum, and multiple planar reconstruction (MPR) were used for image post-processing. The results were compared with those of colonoscopy and pathology.

RESULTSMultislice helical CT colonography detected 54 colorectal carcinomas, 4 adenomas with focal carcinoma, 1 non-Hodgkin's lymphoma (NHL). The lesions' number, size, location, morphology, stricture of intestinal cavity, infiltration, and metastasis were shown satisfactorily by multislice helical CT colonography. Whole colon could be shown in all patients. CT colonography displayed 4 synchronous colonic tumors, 1 ascending colon carcinoma combined with left renal carcinoma among 54 patients with colonic carcinomas. The accuracy of location of CT colonography was 100%. There were 9 cases that CT showed the tumor location was different from the finding of conventional colonoscopy, while all of the CT location were proven exact by operation. CT colonography also displayed the infiltration of serous layer and fatty tissue in 45 cases; 21 cases matched the pathological results in all the 24 cases of suspicious lymph node metastasis, the sensitivity was 87.5%, the specificity was 90.6%; 9 cases hepatic metastasis, 2 ovarian metastasis, and 1 double adrenal gland metastasis.

CONCLUSIONSMultislice helical CT colonography is effective in preoperative diagnosis, location, stage, and making treatment plan of colorectal carcinoma. It can display the portion not seen during colonoscopy and may have an adjunctive role.

Adenocarcinoma ; diagnostic imaging ; Adenoma ; diagnostic imaging ; Adult ; Aged ; Aged, 80 and over ; Colon ; diagnostic imaging ; Colonic Neoplasms ; diagnostic imaging ; Colonography, Computed Tomographic ; methods ; Female ; Humans ; Lymphoma, Non-Hodgkin ; diagnostic imaging ; Male ; Middle Aged ; Tomography, Spiral Computed

Objective To study the factors that influence the ex vivo proliferation of human immunodeficiency virus (HIV) specific CD8 T cells.Methods Three methods to use HIV-specific antigen peptides stimulating the proliferation of virus specific CD8 T cells from HIV patients were compared,and the effect of adding exogenous IL-2 into co-culture on proliferation was evaluated.Results In comparison to directly adding MHC I matched HIV peptides into peripheral blood mononuclear cells (PBMCs) by presence during co-culture or incubating for 2 hours,the more efficient proliferation of HIV-specific CD8 T cells was stimulated by loading HIV peptides on the mixed antigen presenting cells (APCs) that harvested by removing CD8 T cells from PBMCs.Adding IL-2 into co-culture could largely enhance the number of proliferated HIV-specific CD8 T cells but also induced non-specific proliferation of CD8 T cells.Conclusions Stimulation of HIV-specific CD8 T cells by loading HIV-specific antigen peptides on mixed APCs and in absence of IL-2 can specifically induce the efficient proliferation of HIV-specific CD8 T cells by providing both antigen and co-stimulation signals.

Objective:To explore the risk factors of recurrence after resection of thigh soft tissue malignant tumors and the evaluation of the postoperative lower limb function.Methods:A total of 211 patients who underwent thigh soft tissue malignant tumor resection in the Department of Orthopaedics of the second affiliated Hospital of Medical College of Zhejiang University from May 2011 to May 2021 were retrospectively analyzed, including 117 males and 94 females, aged 53 (43, 65) years. There were 59 cases of atypical/well-differentiated liposarcoma, 30 cases of other types of liposarcoma, 33 cases of malignant fibrous histiocytoma, 19 cases of fibrosarcoma, 12 cases of rhabdomyosarcoma, 12 cases of leiomyosarcoma, 9 cases of synovial sarcoma, and 37 cases of others. The tumor involvement of muscle, bone, blood vessel, nerve tissue and intermuscular compartment, postoperative Musculoskeletal Tumor Society (MSTS) functional score and recurrence were recorded. The relationship between anatomical structure and postoperative recurrence was analyzed by Cox logistic regression to determine risk factors; the cumulative recurrence rate of different compartment groups (medial compartment, anterior compartment, and posterior compartment) were compared; The Box plot was used to compare the postoperative function of patients with resection of different anatomical structures (e.g., vastus lateralis, biceps femoris, femur, etc.).Results:A total of 34 out of 211 patients relapsed, with a recurrence rate of 16.1%. The recurrence time ranged from 2.6 months to 91.6 months after operation, with a median recurrence time of 37.0 (18.2, 52.8) months. Three independent risk factors were found to be associated with recurrence, namely: pathological grade [ HR=3.86, 95% CI(1.75, 8.51)], involvement of vastus intermedius [ HR=3.05, 95% CI(1.53, 6.06)], and involvement of vastus medialis [ HR=3.17, 95% CI(1.56, 6.41)]. The recurrence rate of patients with anterior chamber tumor resection was 35.3%, which was higher than that of patients without tumor resection (16.2%), and the difference was statistically significant ( P=0.020). There was no significant difference in recurrence rate between patients with medial chamber tumor resection and patients without tumor resection ( P>0.05). The recurrence rate of patients with posterior compartment tumor resection was 12.3%, which was lower than that of unresectable patients (37.6%), and the difference was statistically significant ( P=0.002).The postoperative MSTS score of 167 patients averaged 26±3.2 points (9-30 points). After intraoperative resection of part of the femur, vastus intermedius, vastus medialis, and rectus femoris, the patient's function was relatively poor [corresponding median MSTS score: 25 (23, 28), 26 (24 28), 26 (24,28), 26(24, 27)]. Conclusion:The risk factors for local recurrence after resection of thigh soft tissue malignant tumors include: pathological grade of the tumor, and whether the vastus intermedius or vastus medialis is involved. Anterior compartment tumors have a higher recurrence rate after surgery. If the tumor involves the above-mentioned anatomical structures, more attention should be paid to the risk of local recurrence after surgery.

Objective:To explore the clinical characteristics of dislocation after resection and reconstruction of tumors involving pelvic area II, and to try to propose intraoperative and postoperative techniques to prevent its occurrence.Methods:From March 2011 to March 2021, 122 patients with resection and reconstruction involving pelvic area II were retrospectively analyzed. Among them, 17 had postoperative dislocation, 32 had pelvic area II, and 31 had pelvic area I+II. There were 40 cases in the pelvic area II+III, and 19 cases in the pelvic area I+II+III. There were 49 female patients and 73 male patients; the mean age was 47 years (9-73 years). The Musculoskeletal Tumor Society (MSTS) score of lower extremity patients after reduction was evaluated, and the clinical characteristics of dislocation, such as dislocation direction, dislocation time and reduction method, were counted and analyzed, and feasible prevention measures were analyzed based on the imaging characteristics after resection and reconstruction. dislocation method.Results:Among the 122 patients, there were 17 cases of dislocation, and the dislocation rate was 13.7%. Among them, 12 cases were anterior dislocation, accounting for 70.6% of anterior dislocation; 5 cases were posterior dislocation, and the proportion of posterior dislocation was 29.4%, difference (χ 2=4.52, P=0.033). There were 12 cases of dislocation within 3 months after operation, accounting for 70.6%; 1 case of dislocation occurred in 5 months after operation, and the other 4 cases of dislocation occurred for more than one year. The MSTS score of the dislocation patients after reduction was 56.1±15.6% (20%-80%). Combined with postoperative imaging examinations, the feasible methods for preventing dislocation include: The anteversion angle of the acetabular cup can be appropriately reduced (0°-10°); The acetabular cup can be appropriately shifted to the rear of the rotation center; Appropriately reducing the anteversion angle of the femoral neck prosthesis, the combination of the three can reduce the occurrence of femoral-acetabular prosthesis impingement; at the same time, the affected limb is controlled in a neutral position with strict nail shoes after surgery to avoid the occurrence of external rotation. Conclusion:The dislocation after tumor resection and reconstruction in pelvic area II is mostly anterior dislocation. Postoperative CT scan of the pelvis shows that the space between the neck of the femoral stem prosthesis and the lower edge of the acetabulum is small, which is prone to impingement. If the anteversion angle of the acetabular cup is appropriately reduced, the acetabular prosthesis is appropriately displaced posteriorly, and the anteversion angle of the femoral neck is reduced by 5°, the possibility of femoral-acetabular impingement can be reduced, thereby reducing the risk of postoperative dislocation.

With the development of high technology, people spend more and more time on screens. Most screens contain a lot of blue light. As one of the important components of visible light, blue light has high energy. It may lead to many ocular diseases, such as myopia, cataract, dry eye, glaucoma and keratitis when eyes exposure to blue light for a long time. At present, the harm of blue light and how to prevent blue light have become a hot topic. Its mechanism mainly includes increasing the photosensitivity of lipofuscin, destroying the mitochondria, lysosome, lens and tear film. According to the mechanism of blue light damage, physical protective measures can be adopted. And the antioxidant, free radical scavengers, anti-inflammatory drugs and gene therapy can be used to protect eye tissue. This paper mainly summarizes the harmful mechanism of blue light to eye and the corresponding prevention and treatment measures.

Objective: To observe the effect of Pinelliae Rhizoma Praeparatum Cum Alumine polysaccharides(PRPCAP)on airway mucus secretion in rats with allergic asthma, in order to study the material basis of the "macromolecule" component of the polysaccharides as the original medicinal materials. Method: The 60 SPF-grade Wistar rats were induced by intraperitoneal injection of chicken ovalbumin (OVA) and aluminum-magnesium adjuvant, except for the normal control group. The OVA solution was aerosolized to establish a rat model of allergic asthma. After successful modeling, the rats were randomly divided into 5 groups, namely allergic asthma model group, positive drug group (montalurast sodium,5 mg·kg^-1), high-dose PRPCAP group (400 mg·kg^-1), middle-dose PRPCAP group (200 mg·kg^-1) and low-dose PRPCAP group (100 mg·kg^-1). The contents of interleukin-4 (IL-4) and interferon-γ (IFN-γ) in serum and bronchoalveolar lavage fluid (BALF) supernatant were determined by enzyme-linked immunosorbent assay (ELISA), and the count of eosinophils (EOS) was detected by BALF sediment. The histopathological changes were observed by hematoxylin-eosin (HE) staining in lung tissue. The mRNA expression of mucin 5AC (MUC5AC) was detected by Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR). Result: Compared with the normal control group, serum IL-4 level in the allergic asthma model group was significantly increased (P<0.01), IFN-γ level was significantly decreased (P<0.01), and EOS counts in BALF were significantly increased (P<0.01). Compared with the allergic asthma model group, the positive drug group, and high, middle, and low-dose PRPCAP groups could significantly reduce the serum IL-4 content (P<0.01), while high and middle-dose PRPCAP groups could significantly reduce the content of IL-4 in BALF (P<0.05, P<0.01), the middle-dose PRPCAP group could significantly increase the content of IFN-γ in serum (P<0.05), EOS counts in BALF were significantly lower in the positive drug group and all of PRPCAP groups (P<0.01). HE staining showed the pathological changes in the lung tissue of the allergic asthma model group and a mitigating effect in the positive drug group and all of PRPCAP groups. Compared with the normal control group, the expression of MUC5AC mRNA in the lung tissue of rats with allergic asthma was significantly increased (P<0.01). Compared with the model group of allergic asthma rats, the positive drug group and all of PRPCAP groups showed significant decrease in MUC5AC mRNA (P<0.01). Conclusion: The "macromolecule" component of polysaccharides in the Pinelliae Rhizoma Praeparatum Cum Alumine may be the material basis for the efficacy of eliminating dampness and eliminating phlegm.

Objective:To investigate the pathological characteristics and the clinical significance of novel coronavirus (2019-nCoV)-infected pneumonia (termed by WHO as coronavirus disease 2019, COVID-19).Methods:Minimally invasive autopsies from lung, heart, kidney, spleen, bone marrow, liver, pancreas, stomach, intestine, thyroid and skin were performed on three patients died of novel coronavirus pneumonia in Chongqing, China. Hematoxylin and eosin staining (HE), transmission electron microcopy, and histochemical staining were performed to investigate the pathological changes of indicated organs or tissues. Immunohistochemical staining was conducted to evaluate the infiltration of immune cells as well as the expression of 2019-nCoV proteins. Real time PCR was carried out to detect the RNA of 2019-nCoV.Results:Various damages were observed in the alveolar structure, with minor serous exudation and fibrin exudation. Hyaline membrane formation was observed in some alveoli. The infiltrated immune cells in alveoli were majorly macrophages and monocytes. Moderate multinucleated giant cells, minimal lymphocytes, eosinophils and neutrophils were also observed. Most of infiltrated lymphocytes were CD4-positive T cells. Significant proliferation of type Ⅱ alveolar epithelia and focal desquamation of alveolar epithelia were also indicated. The blood vessels of alveolar septum were congested, edematous and widened, with modest infiltration of monocytes and lymphocytes. Hyaline thrombi were found in a minority of microvessels. Focal hemorrhage in lung tissue, organization of exudates in some alveolar cavities, and pulmonary interstitial fibrosis were observed. Part of the bronchial epithelia were exfoliated. Coronavirus particles in bronchial mucosal epithelia and type Ⅱ alveolar epithelia were observed under electron microscope. Immunohistochemical staining showed that part of the alveolar epithelia and macrophages were positive for 2019-nCoV antigen. Real time PCR analyses identified positive signals for 2019-nCoV nucleic acid. Decreased numbers of lymphocyte, cell degeneration and necrosis were observed in spleen. Furthermore, degeneration and necrosis of parenchymal cells, formation of hyaline thrombus in small vessels, and pathological changes of chronic diseases were observed in other organs and tissues, while no evidence of coronavirus infection was observed in these organs.Conclusions:The lungs from novel coronavirus pneumonia patients manifest significant pathological lesions, including the alveolar exudative inflammation and interstitial inflammation, alveolar epithelium proliferation and hyaline membrane formation. While the 2019-nCoV is mainly distributed in lung, the infection also involves in the damages of heart, vessels, liver, kidney and other organs. Further studies are warranted to investigate the mechanism underlying pathological changes of this disease.

Objective: To investigate the correlation between CLOCK and BMAL1 genes and MEN2 medullary thyroid carcinoma (MTC). Methods: Thirteen cases with MEN2 MTC and thirteen cases with non-MEN2 MTC were selected who were treated in the Yantai Yuhuangding Hospital between January 2013 and September 2021. Clinical indicators such as blood calcitonin level, tumor diameter and metastatic lymph node of patients were collected. The expression differences of CLOCK and BMAL1 between MEN2 MTC and para-carcinoma tissue as well as between MEN2 MTC and non-MEN2 MTC were detected by immunohistochemistry and qPCR. The correlation between lymph node metastasis and CLOCK or BMAL1 expression was analyzed. Protein-protein interaction (PPI) network analysis combined with qPCR and correlation analysis was used to explore the expression regulation relationship between RET and circadian clock genes. The rhythm disorder of MEN2 cells was verified by lipopolysaccharide cell stimulation experiment after dexamethasone rhythm synchronization. Results: MEN2 MTC exhibited typical RET gene mutation. The mean blood calcitonin level, the tumor diameter and the number of metastatic lymph nodes of patients with MEN2 MTC were higher than those of patients with non-MEN2 MTC (t value was 2.76, 2.53, 2.26, all P<0.05). Immunohistochemical results showed that the expression levels of CLOCK and BMAL1 in MEN2 MTC were higher than those in non-MEN2 MTC, while negatively expressed in para-cancerous thyroid follicle. qPCR displayed that the expression of CLOCK gene in cancer tissues was higher than that in non-MEN2 MTC and para-cancerous tissues (t value was 2.68 and 2.86, all P<0.05); the expression of BMAL1 gene in MEN2 MTC was higher than that in non-MEN2 MTC and para-cancerous tissues (t value was 2.21 and 2.35, all P<0.05). Correlation analysis showed that the expression levels of CLOCK and BMAL1 genes were positively correlated with the number of lymph node metastases in patients with MEN2 MTC (r=0.65, P<0.001; r=0.52, P=0.005). PPI network analysis indicated that the expression of CLOCK gene was positively correlated with the abnormal expression of RET gene (r=0.96, P<0.001). With lipopolysaccharide to stimulate cultured cells in vitro after dexamethasone rhythm synchronization, the expressions of CLOCK and BMAL1 in MEN2 MTC cells (0.47±0.22 and 2.60±1.48) at 12 hours of synchronization were significantly lower than those in para-cancerous tissues (1.70±1.62 and 8.23±2.52), the difference was statistically significant(t=5.04, P=0.007; t=3.34, P=0.029). Conclusion: CLOCK and BMAL1 are correlated with the occurrence and development of MEN2 MTC, and may be potential targets for the development of new therapeutic strategies for MEN2 MTC.
ARNTL Transcription Factors/genetics* ; CLOCK Proteins/genetics* ; Calcitonin ; Carcinoma, Neuroendocrine/genetics* ; Dexamethasone ; Humans ; Lipopolysaccharides ; Lymphatic Metastasis ; Multiple Endocrine Neoplasia Type 2a/genetics* ; Thyroid Neoplasms/surgery*