[Analysis of the complete genome characterization of 11 human astrovirus strains in Shandong Province].

Objective: To study the complete genome characterization of Human Astrovirus (HAstV) in Shandong Province. Methods: Stool samples from acute flaccid paralysis (AFP) surveillance in Shandong Province from 2020 to 2022 were collected, and HAstV nucleic acid was examined by real-time quantitative PCR (qPCR). Next-generation sequencing (NGS) was conducted for the positive samples to obtain complete genome sequences and identify the genotype. Homology comparison and phylogenetic analysis were performed by using BioEdit and Mega software. Results: A total of 667 samples were examined by qPCR, of which 14 were HAstV-positive (2.1%), including HAstV-1 (n=6), MLB1 (n=6), MLB2 (n=1), and VA2 (n=1). The complete genome sequences were obtained from 11 samples. The six HAstV-1 sequences of this study had 98.2% to 99.9% nt similarities with each other and 87.6% to 98.6% with those from other regions. The four MLB1 sequences of this study had 99.1% to 99.9% nt similarities with each other and 92.2% to 99.4% with those from other regions. The VA2 sequence of this study had 96.0% to 96.3% nt similarities with those from other regions. Phylogenetic analysis based on ORF2 region showed that the local HAstV-1 sequences were most closely related to Japanese strains, and had distinct topology with phylogenies based on ORF1a and ORF1b regions. Conclusion: The complete genome sequences of 11 HAstV strains are obtained, and the VA2 complete genome is found.

[Characteristics of genioglossus neuromuscular activity in patients with obstructive sleep apnea during drug-induced sleep].

Objective: To analyze genioglossus (GG) activation responses to the negative pressure of upper airway cavity during awake and different sleep stages in patients with different obstructive sleep apnea (OSA) graduation. Methods: This prospective cohort study started from August 2019 to January 2021, recruited 42 male OSA patients aged from 21 to 59 (38.77±8.42) years. After completing whole night polysomnography (PSG) and upper airway CT, each subject underwent drug-induced sleep with simultaneous monitoring of genioglossal electromyography (GGEMG) and pressure of epiglottis (Pepi). Subjects were divided into three groups of mild OSA(7 males), moderate OSA(12 males), and severe OSA(23 males). The differences in upper airway CT measurements, parameters of GGEMG and Pepi during awake and induced sleep were compared. Statistical analysis was conducted by SPSS 21.0. Results: There was no significant difference in the GGEMG parameters between the mild and moderate groups. In wakefulness, the peak phasic GGEMG of the severe group was higher than the mild group (t=1.249, P=0.025), with no statistically difference in the corresponding Pepi. In the sleep onset, the GGEMG parameters and Pepi in severe group were higher than the other two groups. Linear regression analysis of the maximum GGEMG and maximum Pepi at the end of obstructive apnea (OA) in all moderate plus severe patients (n=35) was shown nonlinear correlation (r=0.28, P=0.694). The airway length of the glossopharyngeal cavity was linearly correlated with the maximum Pepi of OA (r=0.468, R2=0.219, P=0.005). Conclusions: The individual difference of GG activation in OSA patients is related to the severity of the disease (frequency of respiratory events) and negative pressure stimulation. In moderate and severe OSA patients, GG activity is not in harmony with the corresponding negative pressure stimulation, which may be one of the mechanisms leading to the aggravation of OSA.

Uncertainty and influencing factors of the placement error of three-dimensional conformal intensity-modulated radiotherapy for thoracic tumors.

OBJECTIVE: Radiation therapy is an important method for the treatment of chest tumors. This study discussed the placement error of three-dimensional (3D) conformal intensity-modulated radiotherapy in patients with different types of chest tumors and analyzed the relevant influencing factors. PATIENTS AND METHODS: 100 patients with chest tumors diagnosed and treated in our hospital from March 2016 to March 2018 were randomly selected as research subjects, including 42 cases of esophageal cancer, 44 cases of breast cancer, and 14 cases of lung cancer. All patients underwent 3D conformal radiotherapy. The setup errors of patients with esophageal cancer, breast cancer, and lung cancer were detected after 3D conformal radiotherapy. Besides, the influencing factors of 3D conformal for thoracic tumors were analyzed by multiple linear regression analysis. RESULTS: After 3D conformal radiotherapy, the systematic errors of patients with esophageal cancer in X-axis, Y-axis, and Z-axis were -0.10, 1.26 and 0.07, respectively, while the random errors in X-axis, Y-axis, and Z-axis were 1.18, -1.14, and 0.97 respectively. The times for the absolute values of the positioning error with a range of ≤5 mm in X-axis, Y-axis, and Z-axis were 40 (95.24%), 2 (4.76%) and 36 (85.71%), while these with a range of >5 mm in X-axis, Y-axis, and Z-axis were 6 (14.29%), 41 (97.62%) and 1 (2.38%), respectively. For patients with breast cancer, the systematic errors and random errors in X-axis, Y-axis, and Z-axis are -0.19, 1.19, and 0.15, as well as 0.97, 0.02 and 1.29, respectively. The times for the absolute values of the positioning error with a range of ≤5 mm and >5 mm were 41 (93.18%), 3 (6.82%), and 36 (81.82%), as well as 8 (18.18%), 42 (95.45%) and 2 (4.55%), severally. For patients with lung cancer, the systematic errors and random errors in X-axis, Y-axis, and Z-axis were 0.14, 1.42, and 0.15, as well as 1.35, -0.23 and 1.12, respectively. The times for the absolute values of the positioning error with the range of ≤5 mm and >5 mm were 14 (93.33%), 1 (6.67%), and 11 (73.33%), as well as 4 (26.67%), 14 (93.33%) and 1 (6.67%) after 3D conformal radiotherapy. After multiple linear regression analyses, gender and lung volume were the influencing factors of Z-axis setup error, and the lesion location was the influence factor of Y-axis setup error (p<0.05). CONCLUSIONS: There are certain positioning errors in the X-axis, Y-axis, and Z-axis directions of thoracic tumors receiving 3D conformal radiotherapy. Gender, lung volume, and lesion location are all important factors that affect the placement error. The results of this study provide a certain reference for the positioning error of radiation therapy for thoracic tumors, which is conducive to improving the accuracy of radiotherapy and better protecting the surrounding tissues.

DEVELOPMENT OF AN EXTENDED-RANGE BONNER SPHERE SPECTROMETER FOR CHINA INITIATIVE ACCELERATOR-DRIVEN SYSTEM.

To measure the extended-range neutron spectra and calibrate the extended-range neutron dosemeters of the China initiative Accelerator-Driven System (CiADS), an Extended-range Bonner Sphere Spectrometer (EBSS) has been developed. The EBSS was designed based on the PHITS codes, investigating various combinations of materials and diameters of the neutron moderators and the neutron multipliers for extended-range neutrons. Finally, seven polyethylene-only spheres and seven extended-range spheres were selected and subsequently built. The neutron multipliers of the extended-range spheres embedded concentric shells of lead, copper and tungsten. The response functions of the EBSS were analyzed and experimentally validated. It was subsequently tested with 252Cf neutron source and cosmic ray neutron source. The results demonstrate that the EBSS is capable of accurately measuring neutron spectra.

Setup error of electronic portal image device in IMRT for thoracic tumors and its influence.

OBJECTIVE: The aim of this study was to analyze the setup error of the electronics portal image device (EPID) in intensity-modulated radiation therapy (IMRT) for thoracic tumors and the influence on the outward expansion distance of the target area. PATIENTS AND METHODS: A total of 202 patients with chest tumors admitted to our hospital from March 2016 to March 2018 were selected as the observation subjects. All patients were treated with IMRT. The original plan was developed based on the SM90 obtained by the planning target volume (PTV) expansion method, and the new plan was obtained by shifting the isocenter coordinates of the treatment plan according to the positioning error value obtained by EPID. Before the treatment, EPID scans were performed. The electronic radiation field images (ERIs) were registered with the digitally reconstructed radiographic images (DRRs) generated by the treatment planning system using the image registration software, and the setup errors in the X, Y, and Z directions were further measured. The PTV was developed according to ERIs, and the setup error was simulated to obtain the PTV with 95% internal target volume (ITV) reaching the prescribed dose under the condition of a setup error. The outward expansion distance of clinical target volume (CTV) → PTV was calculated. RESULTS: In this experiment, the setup errors in X, Y, and Z directions were (-2.00±1.16) mm, (0.16±1.14) mm, and (-0.55±1.16) mm, respectively. The systematic error in the Z direction was -3.00 mm, and the random error in the X direction was 3.30 mm. The CTV → PTV outward expansion distance was set as 7, 8 and 7 mm in the X direction, Y direction and Z direction, respectively. At this time, under the presence of setup error, the PTV D95 and the ITV V100 in the new plan were (62.23±3.85) Gy and (97.51±1.56) %, respectively, effectively ensuring that 95% ITV of 90% patients reached the prescribed dose. In contrast, the ITV D95 and ITV V100 in the presence of setup error were (56.11±5.26) Gy and (90.15±3.12) %, respectively, at a CTV → PTV outward expansion distance of 5 mm, which could not guarantee that 95% ITV of 90% patients reached the prescribed dose. In the presence of a setup error, the double-lung 5 Gy irradiation of the total heart volume (V5), the double-lung 20 Gy irradiation of the total heart volume (V20), mean lung dose (MLD), mean heart dose (MHD), and D1 cm3 of the new plan increased by 0.89%, 0.29%, 0.13%, 0.06%, and 5 Gy, respectively, compared with the original plan. CONCLUSIONS: In general, the first treatment of radiotherapy in thoracic tumors mostly has a certain degree of setup error, which is most evident in the X direction. When the CTV → PTV outward expansion distance is set at 7, 8, and 7 mm in the X direction, Y direction, and Z direction, respectively, it can effectively ensure that 95% ITV reach the prescribed dose in 90% of patients in the presence of a setup error. EPID helps to achieve the desired effect of radiotherapy, improves the efficacy of radiotherapy, and reduces the side effects caused by radiotherapy errors.

Intra-articular injection of magnesium chloride attenuates osteoarthritis progression in rats.

OBJECTIVE: To explore the effects of Mg2+ on the expression of osteoarthritic markers in human cartilage and synovium tissue explants. To investigate the therapeutic effect of intra-articular injection of Mg2+ in an established rat OA (Osteoarthritis) model of anterior cruciate ligament transection with partial medial meniscectomy (ACLT + PMM). DESIGN: Human cartilage and synovium explants were collected from total knee replacement surgeries and incubated with MgCl2 (20 mmol/L) in vitro. A rat OA model was established by ACLT + PMM surgery in 450-500 g male Sprague Dawley (SD) rats. To select the optimal dose, intra-articular injections of MgCl2 (0.05, 0.5, 5 mol/L) were performed at 4 weeks after the surgery every 3 days for 2 weeks. The effect of optimized MgCl2 was further determined by histology, immunohistochemistry, and quantitative real-time polymerase chain reaction. RESULTS: The expressions of osteoarthritic markers in human cartilage and synovium explants were inhibited by Mg2+in vitro. Immunohistochemical analysis further suggested the inhibitory effects of Mg2+ on the expression of MMP-13 and IL-6 in the human tissue explants. Cartilage degeneration and synovitis in ACLT + PMM rats were significantly improved by intra-articular injections of Mg2+ (0.5 mol/L). Immunohistochemical analysis also showed the regulatory effects of Mg2+ on osteoarthritic markers in both cartilage and synovium in rats, consistent with in vitro results. CONCLUSION: Intra-articular injections of Mg2+ at 0.5 mol/L attenuate the progression of OA in the ACLT + PMM rat model. Such effect was at least in part explained by the promotion of cartilage matrix synthesis and the suppression of synovial inflammation.

[Polysomnographic comparation between dexmedetomidine-induced sleep and natural sleep].

Objective: To compare the parameters of polysomnography (PSG) in sleep structure and respiratory events between dexmedetomidine-induced sleep and natural sleep. Methods: From April 2016 to September 2018, a total of 44 patients with obstructive sleep apnea (OSA) and 3 patients with simple snoring completed PSG monitor both in natural sleep and dexmedetomidine-induced sleep in Department of Otorhinolaryngology Head and Neck Surgery, Beijing Tsinghua Changgung Hospital. The PSG parameters were statistically analysed with SPSS 22.0 software. Results: The average dose of dexmedetomidine was (104.60±27.93) µg, and there was no significant difference between the induced-sleep efficiency and the natural sleep efficiency (82.14%±16.66% vs. 86.50%±9.18%, t=-1.559, P>0.05). There was no rapid eye movement(REM) stages in all 47 subjects and only 1 case of them had non-rapid eye movement(NREM) stage 3 in induced sleep. The percentage of NREM1 in total sleep time was statistically different between the two groups (42.10%±26.71% vs. 17.47%±11.68%, t=5.997, P<0.001),but there was no significant difference in the percentage of NREM2 in total sleep time between the two groups (56.96%±26.0% vs. 62.95%±9.03%, t=-1.521, P=0.135). About respiratory events, there were significant differences in apnea hypopnea index ((46.29±20.23)/h vs. (39.67±25.41)/h), obstructive apnea index (25.20[10.50,45.40]/h vs. 16.20[3.30,35.20]/h) between induced-sleep and natural sleep (t=2.297, Z=-3.008, all P<0.05), these difference were more significant in mild-to-moderate OSA. There were no statistically significant differences in central apnea index (0.00[0.00,2.80]/h vs. 0.40[0.10,1.20]/h), mixed apnea index (0.00[0.00,6.20]/h vs. 0.00[0.00,3.40]/h, hypopnea index (4.20[0.00,3.30]/h vs. 12.00[5.20,17.40]/h), Z=-0.110,-0.508,-1.544, all P>0.05). There were statistical differences in the lowest oxygen saturation (84.77%±7. 59% vs. 80.21%±11.62%, t=2.558, P=0.014). Conclusions: There is no significant difference in sleep efficiency and NREM2 between dexmedetomidine induced sleep and natural sleep.NREM3 sleep is rare induced, but REM sleep is none of all. And dexmedetomidine induced sleep may aggravate obstructive sleep apnea, but not central apnea.

[Analysis of polysomnography results between pre- and post-operation in pediatric obstructive sleep apnea hypopnea syndrome patients].

Objective: To determine the objective effects of adenotonsillectomy on pediatric obstructive sleep apnea hypopnea syndrome (OSAHS) through analyzing the polysomnography (PSG) results between pre and post-operation. Methods: A total of 56 pediatric OSAHS patients were included who underwent adenoidectomy or/and tonsillectomy and completed PSG follow-up from January 1, 2017 to March 31, 2018. All the pediatric patients who underwent adenoidectomy or/and tonsillectomy during the research period were arranged to take a preoperative PSG study. Patients who were diagnosed OSAHS would be encouraged to complete a follow-up PSG study ranged from1 to 3 months after surgery. The parameters of respiration and sleep architecture of PSG were compared and analyzed. The paired student t test was used to compare preoperative and postoperative mean values. The unpaired student t test was used to compare quantitative variables among different groups. The rank sum test was used if the data were abnormal distribution. Results: Totally 238 patients completed preoperative PSG study, 62 patients were diagnosed as pediatric OSAHS, 56 eligible patients finished post-operative PSG. Hypopnea was the majority in all type of respiratory events in 56.45% (35/62) subjects, while central apnea as the majority in 29.03% (18/62) subjects who can also get significant CAI decrease after surgery. However, obstructive apnea as the majority only exist in 14.52% (9/62) subjects. The short-term cure rate of pediatric OSAHS was 85.71% (48/56). The postoperative AHI, MAI, CAI, HI, ODI, LoSpO(2), percentage of stage I sleep and arousal index were significantly decreased, however, the OAI was no statistical decrease. The percentage of stage Ⅱ and rapid eye movement (REM) sleep were significantly increased, while no significant change in percentage of slow wave sleep and sleep efficiency(t=2.32, P=0.017). Conclusions: Pediatric OSAHS manifest different characteristics of respiratory events from that of adults. Adenotonsillectomy can significant decrease respiratory events and improve sleep architecture, however, there are still some patients who can't be completely relieved with adenotonsillectomy.

Amniotic stem cell transplantation therapy for type 2 diabetes: 3 years' follow-up report.

OBJECTIVE: These case series present a retrospective analysis of clinical effects that an allogeneic amniotic cell transplantation can have in treating 4 patients with type 2 diabetes. PATIENTS AND METHODS: Eligible diabetic patients were onset within 3 years, with fasting blood glucose > 7.0 mmol/l. Stem - cells that are collected from neonatal amniotic membrane when a child is born (2 x 107 cells) - are used for the treatment of these patients. Measured by the flow cytometry, stem cells transfer clusters of differentiation CD113 and CD34 at a high level, and through the femoral artery on the left, they are administered into the patient's pancreatic dorsal artery. RESULTS: The major outcome of the study is the improvement of glycemic control. After withdrawing insulin completely, 13.2 months later, there is a reincrease in the level of blood glucose of the patient. After adjusting their treatment again, no one required using insulin again; only need metformin (250-500 mg/day) to control blood glucose levels. CONCLUSIONS: Since this therapy showed no effects on C-peptide, our results suggested one plausible therapeutic strategy improving glycemic control by increasing insulin sensitivity.

Porous tantalum seeded with bone marrow mesenchymal stem cells attenuates steroid-associated osteonecrosis.

OBJECTIVE: Bone marrow mesenchymal stem cells (BMMSCs) have been widely applied in osteonecrosis. However, lack of biomechanical support limited application of BMMSCs. And porous tantalum (PTA) has been identified as a cell-friendly scaffold for bone regeneration. Herein, we aimed to investigate the efficacy of PTA seeded with BMMSCs in the treatment of osteonecrosis. MATERIALS AND METHODS: After the production of PTA seeded with BMMSCs, MTT and GFP were performed to identify the proliferation and adhesion of BMMSCs respectively, which was further examined by scanning electron microscopy (SEM). And real-time PCR was also used to determine mRNA level of osteogenic markers, including Alp, OCN, OPN, Col I and Runx-2 in BMMSCs. Nineteen adult rabbits were applied for building steroid-associated osteonecrosis (SAON) models. Bone formation rate (BFR) and mineral apposition rate (MAR) were determined. And Goldner Trichrome Staining was used in these SAON models, which further confirmed the efficacy of PTA seeded with BMMSCs in SAON. RESULTS: PTA seeded with BMMSCs showed excellent biocompatibility. Additionally, SEM assay showed that BMMSCs adhered tightly and spread fully in the pores of PTA. Next, the expression of ALP and OPN mRNA in BMMSCs were significantly (p < 0.05) higher in the PTA-treated group compared to those in the PTA-untreated group. Furthermore, compared to those treated by only PTA, the dynamic bone formation in rabbits treated by PTA seeded with BMMSCs was significantly increased (p < 0.001) at both week 3rd and week 6th. CONCLUSIONS: The product, PTA seeded with BMMSCs, was successfully produced, and was determined as high efficacy for treatment of steroid-associated osteonecrosis. PTA seeded with BMMSCs may afford a promising option for treating osteonecrosis.

Bisphosphonates induce apoptosis of stromal tumor cells in giant cell tumor of bone.

Giant cell tumour of bone (GCT) is an aggressive primary neoplasm that results in the production of osteolytic lesions. Stromal cells, which form the main neoplastic component of this tumor, regulate the formation of osleoclast-like giant cells that are ultimately responsible for bone destruction. Bisphosphonates prevent bone resorption by inhibiting osteoclast activity and promoting osteoclast apoptosis, and they have been known to induce apoptosis of primary neoplastic cells such as those in breast and prostate cancers. We hypothesized that in bisphosphonates may induce apoptosis not only in osteoclast-like giant cells but also in neoplastic stromal cells of GCT both in vitro and in vivo. Twelve patients with GCT were treated with weekly injections of pamidronate for a period of 6 weeks prior to surgery. GCT specimens were collected at the time of biopsy and during definitive surgery. TUNEL assay was used to evaluate apoptotic DNA fragmentation in cells. In addition, twelve GCT primary cultures from these patients were treated with zoledronate, pamidronate, or alendronate for 48 hours at different doses (3, 30, or 150 microM) and subjected to apoptosis assay by flow cytometry following fluorescent Annexin-V labeling. The results showed that pamidronate significantly induced apoptosis in both osteoclast-like giant cells and stromal tumor cells, in vivo. All three bisphosphonates caused substantial apoptosis of stromal tumor cells in cultures. Zoledronate was the most potent reagent, resulting in an average cell death of 27.41% at 150 microM, followed by pamidronate (22.23%) and alendronate (15.3%). Our observations suggest that these drugs may be considered as potential adjuvants in the treatment of GCT.

Expression of RANKL and OPG mRNA in periodontal disease: possible involvement in bone destruction.

Periodontitis is a complex, multifactorial process affected by bacterial plaque-components and host defense mechanisms. Inflammation of the periodontitium may lead the destruction of the underlying ligament and alveolar bone. Receptor activator of NF-kappaB ligand (RANKL), a novel TNF receptor-related protein is an important factor for osteoclast differentiation and activation. Given osteolysis by osteoclast has been demonstrated in periodontitis, we hypothesized that RANKL expression may be associated with bone destruction in periodontitis. We used semi-quantitative RT-PCR to compare the gene expression of RANKL and osteoprogerin (OPG), a decoy receptor of RANKL, between moderate and advanced periodontitis, and healthy subjects. The level of RANKL mRNA was highest in advanced periodontitis. In contrast, the level of OPG mRNA in both advanced and moderate periodontitis was lower than that in the healthy group. It appears that the ratio of RANKL to OPG mRNA in periodontitis has increased. To determine the localization of RANKL gene transcripts in gingival tissue at the cellular level, in situ hybridization was performed using digoxigenin-labeled specific riboprobes. RANKL mRNA was expressed in inflammatory cells, mainly lymphocyte and macrophages. In addition, proliferating epithelium in the vicinity of inflammatory cells expressed high levels of RANKL mRNA. In short, our data suggest that up regulation of RANKL mRNA in both inflammatory cells and epithelium may be associated with the activation of osteoclastic bone destruction in periodontitis.

The identification and toxicity of the microcystin in the waterbloom obtained from an eutrophied lake.

The water of "J" lake has been seriously eutrophied; concentration of total nitrogen (TN), total phosphorus (TP) and chlorophyll a were all far above the 3rd level of the National Standard of Ground Water of China. The concentration of microcystin (MCYST) of the water at one site (M) was 1865 micrograms/l. There were 2.36 micrograms MCYST-LR per mg dry waterbloom powder.

Intracellular vacuolization caused by the urease of Helicobacter pylori.

Tissue culture cells were exposed to supernatants of Helicobacter pylori for 24 h at 37 degrees C in the presence of various quantities of urea. In the normal human stomach the concentration of urea is less than or equal to 4 mmol/l, and in the presence of this low concentration up to 10% of Vero cells showed intracellular vacuolization. In the presence of 7.5 mmol/l urea, 25% of the cells showed vacuolization. With 30 mmol/l urea, the final pH was 7.6, indicating that vacuolization was not due to change of pH. The first report of vacuolization of tissue culture cells by H. pylori was in a system without added urea but with concentrated bacterial supernatant; 30% of H. pylori strains demonstrated a cytotoxic effect. In those experiments fetal calf serum was used; it contains 6 mmol/l urea but was used at a concentration of 10%. A urease inhibitor, acetohydroxamic acid, caused a 75% drop in the number of cells showing vacuolization, and ammonia caused vacuolization. Thus the urea of H. pylori probably causes this vacuolization.

Etiologic studies of epidemic hemorrhagic fever (hemorrhagic fever with renal syndrome).

Two strains of epidemic hemorrhagic fever (EHF) virus were isolated from the lung tissues of Apodemus agrarius mice that were captured in an area where EHF is endemic. The strains were isolated by passages in A. agrarius mice from a nonendemic area. Identification of the isolates by usual procedures was confirmed by repeated blind tests with coded sera. Contamination with certain known viruses such as reovirus, adenovirus (types 3 and 7), and other pathogens, such as murine typhus rickettsiae and Leptospira, which may be naturally present in wild rodents, appeared to have been ruled out. The antigen slides made from these isolates are in use in the specific diagnosis and seroepidemiologic studies of EHF. The first successful application is the serodiagnosis of a mild type of hemorrhagic fever that occurs with characteristic epidemiologic features in certain provinces of China.