Objective:To investigate a rapid protocol for the acquisition of a three-dimensional (3D) image of corneal nerve distribution and its various parameters.Methods:Four SPF female C57BL/6 mice were selected and four corneal samples with complete limbi were obtained using a dissecting microscope after the sacrifice of mice euthanized by ether.After conventional fixation, permeabilization, and immunostaining by an anti-β-Ⅲ tubulin fluorescent-conjugated antibody, a whole-nerve image of the whole-mount cornea was captured under a 60X oil lens using a scientific complementary metal-oxide-semiconductor detector and a high-resolution deconvolution microscope.The 3D image of the corneal nerve fiber was obtained after 3D deconvolution processing, Z-axis data projection, and automatic stitching using the self-contained image processing software of the microscope system.The corneal nerve density in different areas was analyzed using the automatic detection mode in the Filament Tracer module and the manual Autopath module of the interactive microscopic image analysis software Imaris.The use and care of animals complied with the statement of the Association for Research in Visual and Ophthalmology, and the study protocol was approved by an Experimental Animal Welfare Committee of Jinan University (No.JN-A-2002-01).Results:It was found that stromal nerve fibers in a dense network entered the Bowman membrane near the limbus, and branches of stromal nerve fibers formed subbasal nerve plexus, which stretched toward the center of the cornea to form a dense neural network-like structure and converged into a vortex-like structure at the apex of the cornea.Some subbasal nerves entered the epithelial layer vertically and some branches of nerve endings were found.Through the automatic detection mode of Imaris software, a gradual increase of the density from (2 488.88±282.84)μm/μm 2 at the limbus to (5 766.66±298.55)μm/μm 2 at the center of the cornea of the subbasal nerve branches, and a decrease of the density from (40.99±0.99)μm/μm 2 at the limbus to (34.57±1.28)μm/μm 2 at the center of the stromal nerves were found.It was also found that the stromal nerves at the limbus entered the Bowman membrane for about 151 μm and then began to branch to form subbasal nerves. Conclusions:The high-resolution deconvolution microscope system can be used to study the 3D distribution of the whole corneal nerve.Additionally, Imaris can be used for obtaining various parameters of the corneal nerves automatically and quickly.

Objective    To evaluate the clinical effects of segmentectomy versus lobectomy under single utility port video-assisted thoracic surgery on inflammatory factors and immune cells in peripheral blood of non-small cell lung cancer patients, and to analyze the effect of changes of postoperative inflammatory factors and immune cells on the prognosis of the patients. Methods    The clinical data of 256 patients who underwent segmentectomy or lobectomy under single utility port video-assisted thoracic surgery for non-small cell lung cancer in the First Affiliated Hospital of Hebei North University from January 2016 to October 2020 were retrospectively collected. According to the operation method, they were divided into a segmentectomy group (126 patients with 79 males and 47 females at an age of 63.4±6.2 years) and a lobectomy group (130 patients with 91 males and 39 females at an age of 62.9±5.6 years). The change of inflammatory factors (C reactive protein, interleukin-6, interleukin-8, tumor necrosis factor-α) and immune cells (CD4+T cells, CD8+T cells and natural killer cells) were recorded and analyzed before operation (T0) and 1 day (T1), 3 days (T2), 7 days (T3), 1 month (T4) after the operation between the two groups. According to postoperative recurrence situations, they were divided into a recurrence group and a non-recurrence group, multivariate logistic regression analysis was used to analyze the relationship between the change of postoperative inflammatory factors, immune cells, and the prognosis of patients with non-small cell lung cancer. Results    (1) There was no statistical difference in sex ratio, underlying diseases, body mass index, levels of preoperative inflammatory factors or immune cells between the two groups (all P>0.05). (2) The changes of postoperative inflammatory factors in the segmentectomy group were significantly less than those in the lobectomy group at T1-T3 (all P<0.05), and the changes of postoperative immune cells in the segmentectomy group were significantly less than those in the lobectomy group at T1-T4 (all P<0.05). (3) The changes of postoperative inflammatory factors and immune cells on postoperative day 3 in the recurrence group were significantly more than those in the non-recurrence group (all P<0.05). (4) Multivariate logistic regression analysis showed that the changes of postoperative inflammatory factors and immune cells on postoperative day 3 may be the risk factors for postoperative recurrence and metastasis in patients with non-small cell lung cancer (all P<0.05). Conclusion    Single utility port video-assisted thoracic surgery segmentectomy for the treatment of non-small cell lung cancer can reduce the inflammatory response and protect body's immune function, and the change of postoperative inflammatory factors and immune cells in postoperative day 3 may be the risk factors for postoperative recurrence and metastasis in patients with non-small cell lung cancer.

Radiation-induced lung injury is a common complication of thoracic malignant tumor radiotherapy and severe nuclear accident injury. Currently, there is no effective treatment on this injury. Mesenchymal stem cells (MSCs) are a group of cells with multi-directional differentiation potential and they can protect lung tissue from radiation damage by homing to the injured site and differentiating to the damaged tissues, secreting cytokines and immune regulation. Further, the genetically modifying mesenchymal stem cells have not only the main characteristics of MSCs, but also can efficiently and stably express or knock down a certain of target genes, thereby enhancing or reducing the sensitivity of mesenchymal stem cells to various physiological stimulus and enhancing its therapeutic effect in radiation-induced lung injury, providing new ideas and new strategies for clinical treatment. This paper reviewed the relevant research progress in recent years.

Objective To explore the key initiatives and effective methods for preparing the constructions of emergency-oriented hospitals under COVID-19 pandemic. Methods The wartime mechanism was strengthened by adhering to unified leadership, trengthening the top-level design and clarifying the division of responsibilities. Objective management was used as a means to take into account the key of personnel allocation and training, prevention and control of hospital infection, transformation of contagious ward, logistic support, equipment and material supply and construction of system and process. Results The preparations and constructions of the emergency-oriented hospitals were completed in 72 hours，which passed the acceptance and inspections from infection control experts，who appraised our work to be “the highest in difficulty, the fastest in project progress and the highest in quality". Totally, upon to the preparations，14 medical teams were set up and the layout process reestablishment of 14 wards was completed, the installation and preparation of nearly 10000 sets/pieces of medical equipment and medical materials were completed as well and more than 80 work systems and process systems for 9 major modules were established. Conclusion The preparations and constructions of emergency-oriented hospitals should be performed upon the thorough implementation of the decisions and arrangements by the municipal Party committee and the municipal government, insisting on the wartime thinking and establishment of high-quality management team and effective goal management focusing on details and actual needs of medical staff.

Objective To evaluate the effect of an inverse planning simulated annealing (IPSA) in the treatment of cervical cancer with combined intracavitary and interstitial three-dimensional brachytherapy.Methods A total of 60 patients with locally advanced cervical cancer who received both external beam radiotherapy and combined intracavitary and interstitial brachytherapy in our hospital from October 2016 to July 2018 were enrolled.Patients were divided into four groups with 15 patients each according to the number of needles applied (1,2,3,and 4 needles,respectively).Dosimetric distributions were optimized with both Graphical optimization (GRO) and IPSA.Paired t-test was applied to compare the dosimetric differences between plans optimized with GRO and IPSA.Results The Dg0 and V100 of IPSA plans were higher than those of GRO (t=-4.742,-4.823,P＜0.05),while the conformity index (CI) and conformal index (COIN) were slightly lower than those of GRO plans (t=9.642,8.783,P＜0.05).No significant difference in the V150,V200,V300 between IPSA and GRO (P＞0.05) was observed.There was also no significant difference in the D2cm3 of bladder and rectum between IPSA and GRO (P＞0.05).The difference of Dg0 between IPSA and GRO was increased as the number of implanted needles increased,which increased from 4 cGy to 14 cGy as the number of needle increased from 1 to 4.The difference of V100 between GRO and IPSA was also increased as the number of needle increased.Conclusions In the treatment of cervical cancer with combined intracavitary and interstitial threedimensional brachytherapy,IPSA plan could improve the target coverage(D90,V100)without increasing the dose to the OARs and high dose region in the target compared with GRO.With the numbers of needles increased,the advantage of IPSA increased in terms of target coverage.

Objective To establish a stably overexpressing miR-31 transgenic mouse and detect the expression of miR-31 in the organs and tissues,and to provide qualified tool mice with overexpression of miR-31 in vivo. Methods The miR-31 overexpression vector was constructed by Gateway cloning technology. The vector was injected into fertilized ovum by DNA microinjection technology,then transferred to the pseudopregnant mice and waited for eutocia. Newborn mouse tail DNA was extracted and PCR and agarose gel electrophoresis were performed to identify the positive miR-31 transgenic mice. microRNA was extracted from the organs and tissues of miR-31 transgenic mice and the expression of miR-31 was de-tected by RT-PCR. The expression of Nestin and number of neural stem cells in the nervous system were compared in the positive and WT mice. Results The miR-31 transgenic mice were constructed successfully and bred more than 14 genera-tions in barrier environment. Expression of miR-31 was increased in major organs and tissues. The expression of Nestin and the number of neural stem cells in the positive mice were higher than those in the wild type mice. Conclusions MiR-31 overexpressing transgenic mice are constructed by Gateway cloning technology and the expression of miR-31 is stable in sub-sequent generations. The number of neural stem cells in the nervous system is higher than that in wild-type mice. The miR-31 overexpressing transgenic mice can be a good tool for experimental research of the function of overexpressed miR-31 in vivo and the treatment of nervous system diseases.

Multiple institutions at home and abroad have established and issued the standards,guidelines,criteria or reports of the quality assurance (QA) and quality control (QC) of radiotherapy.At present,no institutions in China have implemented systematic classification of QA-and QC-related criteria of radiotherapy.Complete standard system of QA and QC of radiotherapy should consist of clinical and physical technologies.Physical technology includes equipment,technological flow and radiation protection.The QA and QC of physical technology play a pivotal role in tumor radiotherapy.This paper introduces relevant criteria of QA and QC of radiotherapy at home and abroad.

Objective To evaluate the effect of dexmedetomidine on endoplasmic reticulum stress during intestinal ischemia-reperfusion (I/R) in mice.Methods Thirty SPF healthy male C57BL/6 mice,aged 8 weeks,were divided into 3 groups (n=10 each) using a random number table method:sham operation group (S group),intestinal I/R group (I/R group) and dexmedetomidine group (DEX group).The superior mesenteric artery was only isolated but not clamped in S group.The model of intestinal I/R injury was established by clamping superior mesenteric artery for 20 min followed by 24 h of reperfusion in I/R group and DEX group.Dexmedetomidine 25 μg/kg was intraperitoneally injected at 30 min before ischemia in DEX group,while the equal volume of normal saline was given instead of dexmedetomidine in S group and I/R group.Mice were sacrificed at 24 h of reperfusion,and small intestinal tissues was obtained for examination of the pathological changes and ultrastructure of intestinal epithelial cells and for determination of cell apoptosis,expression of CCAAT-enhancer binding protein homologous protein (CHOP),transcription factors (ATF4),and X-4 box binding protein 1 (XBP1) mRNA (by real-time polymerase chain reaction),and expression of CHOP,Bcl-2,Bax and caspase-3 in intestinal tissues (by Western bolt).The apoptosis index (AI) and ratio of Bcl-2 to Bax were calculated.Intestinal damage was assessed and scored according to Chiu.Results Compared with S group,Chiu's score and AI were significantly increased,the expression of CHOP,ATF4 and XBP-1 mRNA,CHOP,Bax and caspase-3 was up-regulated,and the expression of Bcl-2 was down-regulated,and Bcl-2/Bax ratio was decreased in I/R group and DEX group (P＜0.05).Compared with I/R group,Chiu's score and AI were significantly decreased,the expression of CHOP,ATF4 and XBP-1 mRNA,CHOP,Bax and caspase-3 was down-regulated,and the expression of Bcl-2 was up-regulated,and Bcl-2/Bax ratio was increased in DEX group (P＜0.05).Conclusion The mechanism by which dexmedetomidine reduces intestinal I/R injury may be related to regulating endoplasmic reticulum stress and inhibiting cell apoptosis in mice.

Objective To investigate the effect of esmolol pretreatment on Toll like receptor-4 (TLR4)/nuclear factor-kappa-B (NF-кB) pathway in rats with repeated cerebral ischemia/reperfusion (IR) injury. Methods Forty-eight adult male Sprague-Dawley rats were randomly allocated into sham-operated group, IR group and esmolol group (n=16). Rats in the IR group and esmolol group were injected intravenously with esmolol at a dose of 200 g/(kg?min) or normal saline for one h before surgery, and then, bilateral common carotid arteries were clipped to establish the repeated IR injury models. Bilateral common carotid arteries in rats of sham-operated group were only isolated but not clipped, and injected intravenously with normovolemic normal saline for one h. Learning and memory abilities of rats were measured by Morris water maze test before, and one, 3 and 7 d after surgery. Rats were euthanized and hippocampus tissues were excised. The wet to dry (W/D) ratio of the hippocampus was tested. The permeability of blood-brain barrier was determined by Evans blue (EB) method. The levels of tumor necrosis factor-alpha (TNF-α), interleukin (IL)-6 and IL-1β in the hippocampus were tested by enzyme linked immunosorbent assay (ELISA). The NF-кB p65 and TLR4 mRNA expressions in the hippocampus were determined by reverse transcription-polymerase chain reaction (RT-PCR). The NF-кB p65 and TLR4 protein expressions in hippocampus were detected by Western blotting. Results As compared with those in the sham-operated group, the escape latency was significantly prolonged and swimming distance was signficantly longer in rats of IR group one, 3 and 7 d after surgery (P<0.05), the W/D ratio of the hippocampus and the content of EB in brain tissues were significantly increased in the IR group (P<0.05), the levels of TNF-α, IL-6 and IL-1β in hippocampus were significantly increased in the IR group (P<0.05), and the NF-кB p65 or TLR4 mRNA and protein expressions in the hippocampus of IR group were statistically higher (P<0.05). As compared with IR group, the escape latency and swimming distance of rats in the esmolol group were significantly shortened one, 3 and 7 d after surgery (P<0.05), the W/D ratio of the hippocampus and content of EB in brain tissues of esmolol group were significantly decreased (P<0.05), the levels of TNF-α, IL-6 and IL-1β in the hippocampus of esmolol group were signficantly lower(P<0.05), and the NF-кB p65 or TLR4 mRNA and protein expressions in hippocampus of esmolol group were statistically lower in the esmolol group (P<0.05). Conclusion Esmolol preconditioning can alleviate cerebral injury and improve learning and memory abilities of rats with repeated cerebral IR injury, which may be involved in alleviating inflammation and suppressing TLR4/NF-кB pathway.

Arsenic trioxide (ATO) is used as a chemotherapeutic agent for the treatment of acute promyelocytic leukemia. However, increasing drug resistance is reducing its efficacy. Therefore, a better understanding of ATO resistance mechanism is required. In this study, we established an ATO-resistant human epidermoid carcinoma cell line, KB/ATO, from its parental KB-3-1 cells. In addition to ATO, KB/ATO cells also exhibited cross-resistance to other anticancer drugs such as cisplatin, antimony potassium tartrate, and 6-mercaptopurine. The arsenic accumulation in KB/ATO cells was significantly lower than that in KB-3-1 cells. Further analysis indicated that neither application of P-glycoprotein inhibitor, breast cancer resistant protein (BCRP) inhibitor, or multidrug resistance protein 1 (MRP1) inhibitor could eliminate ATO resistance. We found that the expression level of ABCB6 was increased in KB/ATO cells. In conclusion, ABCB6 could be an important factor for ATO resistance in KB/ATO cells. The ABCB6 level may serve as a predictive biomarker for the effectiveness of ATO therapy.