Objective To investigate antigen optimization of Shisa like protein 1 (SHISAL1) for preparing mouse anti-human SHISAL1 polyclonal antibody and to identify the specificity of the prepared antibody. Methods Bioinformatics was employed to predict the antigenic epitope region of SHISAL1 protein, and then a polypeptide composed of amino acid residues from the site of 28 to 97 of SHISAL1, termed SHISAL1-N, was selected as the antigen. The coding region of SHISAL1-N was cloned by molecular cloning technique, and then it was inserted into pET-28a to generate pET28a-SHISAL1-N recombinant plasmid. The two recombinant plasmids pET28a-SHISAL1-N and pET28a-SHISAL1 were transformed into BL21 (DE3) bacteria and induced to express by IPTG. The two proteins were purified and immunized to female Kunming mice, respectively. The specificities and sensitivities of the acquired antibodies were detected by Western blot analysis, immunoprecipitation and immunofluorescent cytochemical staining. Results pET28a-SHISAL1-N recombinant plasmid was successfully constructed, and the two fused proteins, SHISAL1 and SHISAL1-N, were induced to express. Moreover, two types of SHISAL1 mouse polyclonal antibodies, derived from SHISAL1-N and SHISAL1 antigens, were obtained. Western blot results showed that the antibody prepared from SHISAL1 antigen was less specific and sensitive compared with the antibody prepared from SHISAL1-N antigen which could specifically identify different endogenous SHISAL1 protein. Immunoprecipitation results showed that SHISAL1-N antibody could specifically pull down SHIISAL1 protein in hepatocellular carcinoma cells and immunofluorescence results demonstrated that SHISAL1-N antibody could specifically bind to SHISAL1 protein in the cytoplasm. Conclusion We have optimized the SHISAL1 antigen and prepared the mouse anti-human SHISAL1 polyclonal antibodies successfully, which can be used for Western blot analysis, immunoprecipitation and immunofluorescence cytochemical staining.
Animals ; Female ; Humans ; Mice ; Antibodies ; Antibody Specificity ; Blotting, Western ; Cloning, Molecular ; Epitopes/genetics*

Objective:To evaluate the efficacy of proximal humerus internal locking system (PHILOS) via the minimally invasive plate osteosynthesis (MIPO) in the treatment of proximal humeral fractures.Methods:This retrospective study analyzed 30 elderly patients with proximal humeral fracture who had been treated by PHILOS via MIPO from September 2016 to March 2020 at Department of Orthopedic Surgery, People’s Hospital of Zhuxi County. They were 19 females and 11 males with an average age of 60.96 years (from 45 to 80 years). All patients were treated by closed reduction. After fracture reduction was confirmed by fluoroscopy, a minimally invasive incision was made below the acromion, with a couple of suture wires reserved at the rotator cuff attachments. A PHILOS plate was inserted at 4 mm lateral to the intertubercular sulcus, with the suture wires passing through the proximal suture holes on the PHILOS. After a lag screw was first screwed up into the compression hole on the PHILOS plate, the crossing suture wires were tightened up to resist the rotator cuff stress and maintain the internal inclination of the humeral head. Kirschner wires were used to temporarily stabilize the reduction. After satisfactory reduction and fine plate positions were confirmed by fluoroscopy, locking nails were screwed up. The internal inclination of the affected humeral head was compared between preoperation and the last follow-up. The therapeutic efficacy was evaluated by the shoulder Neer scoring system, and the visual analogue scale (VAS) pain scores and complications were recorded at the last follow-up.Results:All the 30 patients were followed for 6 to 18 months (average, 12 months). There was no incision infection, neurovascular injury, or internal fixation failure. Anatomical reduction was achieved in 25 patients and functional reduction in 5. The inclination of the humeral head was significantly improved. The VAS scores at the last follow-up averaged 1.9. Follow-up X-ray examination showed that bony union was achieved after 6 to 18 months (average, 9 months) for all patients. At the last follow-up, the inclination angle of the affected humeral head was 130°±5°, significantly improved compared with the preoperative 90°±11.2° ( P<0.05). All patients had good functional recovery of the shoulder. The efficacy was, according to the Neer shoulder scores at the last follow-up, excellent in 22, good in 6 and fair in 2 cases. Conclusions:Treatment of proximal humeral fractures using PHILOS via MIPO technique is suitable for patients with osteoporotic fracture, and may lead to fine therapeutic efficacy.

Objective:To evaluate the effect of intestinal carbapenem-resistant Enterobacteriaceae (CRE) active screening combined with enhanced intervention in the prevention and control of nosocomial infection in patients admitted to the hematological ward.Methods:Patients who were admitted to the Department of Hematology in a tertiary-care general hospital from March 1, 2017 to December 31, 2019 and underwent chemotherapy or immunosuppressive therapy comprised the intervention group. They were screened for intestinal CRE at least thrice. From December 1, 2016 to February 28, 2017, patients who underwent chemotherapy or immunosuppressive therapy without active intestinal CRE screening in the Department of Hematology formed the control group. Both the patient groups were monitored for CRE infection in real time. The χ2 test was used to compare the changes in the CRE infection rate and mortality in high-risk patients before and after the active screening. Results:During the intervention period, the CRE colonization rate of patients was 16.46% (66/401) ; in terms of disease distribution, the colonization rate of acute leukemia was the highest 23.03% (26/113) . Of the 66 colonized patients, 27 (40.9%) patients were identified as positive for CRE at the first screening, 15 (22.7%) were identified at the time of the second screening, and the remaining 24 (36.4%) were identified at the third or subsequent screening; Carbapenem-resistant Klebsiella pneumoniae (CRPK) strains were dominant among the pathogens, accounting for 54.55% (36/66) . During the active screening period, the CRE infection rate (2.49%) and mortality rate (50.00%) of high-risk patients were significantly lower than those of the controls (11.30% and 69.23%, respectively) . The pathogens of 10 CRE infection patients during the intervention period were exactly the same as the previous active screening pathogens, and the coincidence rate was 100%.Conclusion:The CRE colonization rate was the highest in patients with acute leukemia who were admitted in the hematology wards. CRPK is the main pathogen of CRE colonization, infection, and death. Increasing the frequency of screening can significantly raise the positive rate of screening, Active screening can effectively reduce the incidence and subsequent mortality of CRE in high-risk patients admitted in the hematological wards. High coincidence rate between CRE screening positive pathogens and subsequent CRE infection pathogens. Intestinal CRE screening can serve as an indicator of CRE bloodstream infection in patients with hematological diseases as well as provide information for antibiotics therapy.

RTOG 0617 trial has indicated that no benefit can be obtained in the overall survival of locally advanced non-small cell lung cancer patients by improving the prescribed dose, which promotes the adjustments to the strategies of dose escalation. Currently, multiple studies have been designed to explore more effective approaches to boost dose, such as dose boosts based on increased ¹⁸FDG-uptake regions, simultaneous integrated boost intensity-modulated radiotherapy and modulation of dose fractions, which have achieved a series of progress. The widespread application of PET-CT and intensity-modulated radiotherapy offers broad space for the dose escalation and optimization.

Objective Investigate the relationship between gross tumor volume (GTV)-related factors including GTV-T volume,the maximum thickness of the esophageal lesion plane and GTV-T volume/length(GTV-T volume divided by the length of the lesion calculated by the number of GTV-T layers) and the locoregional failure of radical intensity-modulated radiation therapy (IMRT) for esophageal carcinoma.Methods A total of 133 patients with esophageal cancer undergoing radical IMRT were enrolled.The factors related to GTV-T including GTV-T volume,the maximum thickness of the esophageal lesions,GTV-T volume/length were calculated.The relationship between GTV-T related factors and local recurrence of tumors was retrospectively analyzed.Results There was positively linear association between the locoregional failure rate of GTV-T and the volume of GTV-T.The volume of GTV-T tumor was 36 cm3,the maximum wall thickness was 2.5 cm,and the GTV-T volume/length was calculated as 5.3 cm2.These critical values could be utilized to predict the risk of locoregional failure of IMRT for esophageal carcinoma.Conclusions The GTV-T factors can be adopted to predict the local control and the risk of locoregional failure of radical IMRT for esophageal carcinoma to certain extent.

OBJECTIVE： To provide reference for improving the quality standard of Pheretima. METHODS： The contents of hypoxanthine and inosine in medicinal material samples were determined by HPLC. HPLC fingerprint of Pheretima was established according to “Similarity evaluation system for TCM chromatogramtic fingerprint” （2012 edition） software， and similarity evaluation was conducted. The determination was performed on Purospher STAR RP-18 endcapped with mobile phase consisted of methanol-water （gradient elution） at the flow rate of 1 mL/min. The detection wavelength was 248 nm， and the column temperature was set at 30 ℃. The sample size was 20 μL. RESULTS： The results of methodological investigation of content determination showed that the linear range of hypoxanthine and inosine were 1.58-31.6 μg/mL （r＝0.999 9）， 5.52-110.4 μg/mL（r＝0.999 8）， respectively. limits of quantify were 0.316， 0.552 μg/mL， respectively； limits of detection were 0.158， 0.110 μg/mL， respectively； RSDs of precision， stability （24 h） and repeatability tests were all less than 2.0％ （n＝6）. Average recovery rates were 103.0％ （RSD＝1.7％， n＝6） and 101.2％ （RSD＝1.2％， n＝6）， respectively. HPLC fingerprint for 15 batches of samples were established， and 8 common peaks were identified. The similarity of HPLC fingerprint of 14 batches of sample with control fingerprint R was higher than 0.900. CONCLUSIONS： The established method for content determination of hypoxanthine and inosine and HPLC fingerprint of Pheretima are simple， accurate and reproducible， and can be used for quality control of Pheretima.

Objective To investigate the application of 3D reconstruction system in the clinical departments of the hospital.Methods 3D reconstruction system had its architecture,main features,working flow and principle of auto preprocessing software introduced,and then applied to auto preprocessing of PACS images to realize auto reconstruction,which had the function of 3D post processing.Results 3D reconstruction system gifted the doctor in clinical departments with the access to the images and after treatment,and changed the traditional working mode in imaging department.Conclusion Fusion imaging has 3D reconstruction as the main technique,which eliminates the deficiency in reading radiological images and innovates medical service mode.

Objective To investigate the clinical value of vacuum pad and body film fixation in radiotherapy for thoracic and abdominal tumors. Methods A total of 240 patients with thoracic and abdominal tumors who were treated with radiotherapy were randomly selected and divided into group A (simple vacuum pad fixation,60 patients),group B(simple body film fixation with unimproved solid plate, 60 patients),and group C(vacuum pad and body film fixation with improved solid plate,120 patients).The difference between groups were analyzed with single variance analysis method. Results The setup error was small in group C and large in groups A and B. There were significant differences in the setup error between the three fixation methods(P=0.000). Conclusions A combination of vacuum pad and body film fixation is better than vacuum pad or body film fixation in radiotherapy for thoracic and abdominal tumors. The combination method has many benefits, including simple and convenient operation, comfortable and repeatable body fixation,reduced artificial errors,and improved positioning precision.

Objective To analyze the effects of gross tumor volume(GTV-T)and positive lymph node volume(GTV-LN)on the prognosis of radical concurrent chemoradiotherapy for esophageal squamous cell carcinoma(ESCC). Methods A total of 79 patients with stage N1ESCC undergoing radical radiotherapy in our hospital from 2012 to 2015 were enrolled as subjects. GTV-T and GTV-LN were calculated by the Pinnacle39.0 treatment planning system. The receiver operating characteristic(ROC)curves were used to evaluate the value of the GTV-LN/GTV-T ratio in the prediction of local recurrence(LR)and distant metastasis(DM)of ESCC. Results The median follow-up time was 17.2 months in all patients. The ROC curves were made using the GTV-LN/GTV-T ratio. The optimal cut-off values of GTV-LN/GTV-T ratio for predicting the risk of LR and DM were 0.34 and 0.59, respectively. The statistical analysis revealed that the LR rates were 50% and 8% in patients with GTV-LN/GTV-T ratios of<0.34 and ≥0.34, respectively(P<0.01), while the DM rates were 11% and 43% in patients with GTV-LN/GTV-T ratios of<0.59 and ≥0.59, respectively(P= 0.003). Conclusions The GTV-LN/GTV-T ratio may be a predictor of LR and DM in patients with ESCC. Further studies on the GTV-LN/GTV-T ratio may help to make personalized chemoradiotherapy strategies for patients with ESCC.