Objective To investigate the interference causes of indwelling gastric tube for patients undergoing laryngeal tumor surgery,and explore the appropriate clinical nursing countermeasures.Methods 60 cases of laryngeal cancer patients with indwelling gastric tube were divided into the control group and the observation group with 30 cases in each group.The control group used indwelling gastric tube with routine nursing,while in the observation group,the influencing factors for indwelling gastric tube were analyzed and per-tinent neasures were adopted timely.The satisfaction degree of patients,incidence of postoperative compli-cations,hospitalization time and effective rate of treatment were compared.Results The factors influ-encing use of indwelling gastric tube were tracheal extubation by patients,trachea cannula exodus,pipe blockage and postoperative complications.After timely expectant treatment and positive nursing,the nutri-tion intake and postoperative recovery were not affected.The related indexes of the observation group were better than those of the control group.Conclusions Patients after laryngeal cancer surgery need longtime indwelling gastric tube.Measures such as strengthening of education and propaganda,close observation of patients′ condition and timely expectant treatment should be given in order to ensure the normal use of gastric tube and reach desirable treatment effect.

Objective: Surfactant protein A (SP-A) is protein that appears to play an important role in mammalian first-line host defense. The objective of this study was to immunolocalize SP-A in human sinonasal tissue. Method:Eleven cases of allergic rhinitis, fifteen cases of polyp and seven cases of normal middle turbinate were studied with immunohistochemistry and immunofluorescence method to detect the expression of SP-A. Result:The expression of SP-A in allergic rhinitis and polyp were dramatically higher than that in controls(P<0.05), and there was no remarkable difference in the expression of SP-A between allergic rhinitis and polyp(P>0.05). The result demonstrated that SP-A was positivly correlated with eosinophils within the basement membrane of epitheli-um(R=0.81,0.55). In the result of immunofluorescence, there was significantly higher expression SP-A in nasal mucosa of allergic rhinitis and nasal polyp than that in control group(P<0.05). Conclusion:SP-A is likely to play key roles in the inflammatory reaction process of allergic rhinitis and polyp. Its secretion in the upper airway indicates that future studies may allow manipulation of this protein and development of novel treatments for sinonasal pathology.

OBJECTIVE To study the significance by analyzing the expression of human protection of telomeres1(POT1)and telomeric repeatbinding factor-2(TRF2)in the tissues of laryngneal squamous cell carcinoma(LSCC)and in polyp of vocal cord tissues. METHODS The expression of POT1 and TRF2 in 20 tumor samples and 19 polyp of vocal cord were determined by the indirect immunofluorescence method. The results were scored and analyzed statistically. RESULTS The positive expression rates of POT1 and TRF2 in tumor samples was 65.00% and 70.00% respectively. There were no positive expression of POT1 and TRF2 in polyp of vocal cord. The positive expression of POT1 was higher in poorly differentiated laryngeal carcinoma than that in well-differentiated and moderately differentiated laryngeal carcinoma(Chi-square test with contingency table, P0.05). CONCLUSION The expression of POT1 and TRF2 in laryngneal squamous cell carcinoma were remarkable, POT1 and TRF2 may play a critical role in tumorigenesis of laryngneal squamous cell carcinoma. There was statistical significant difference for degrees of POT1 expression in different tumor histological grades.

OBJECTIVE: Surfactant protein A (SP-A) is protein that appears to play an important role in mammalian first-line host defense. The objective of this study was to immunolocalize SP-A in human sinonasal tissue. METHOD: Eleven cases of allergic rhinitis, fifteen cases of polyp and seven cases of normal middle turbinate were studied with immunohistochemistry and immunofluorescence method to detect the expression of SP-A. RESULT: The expression of SP-A in allergic rhinitis and polyp were dramatically higher than that in controls (P < 0.05), and there was no remarkable difference in the expression of SP-A between allergic rhinitis and polyp (P > 0.05). The result demonstrated that SP-A was positively correlated with eosinophils within the basement membrane of epithelium (R = 0.81, 0.55). In the result of immunofluorescence, there was significantly higher expression SP-A in nasal mucosa of allergic rhinitis and nasal polyp than that in control group (P < 0.05). CONCLUSION SP-A is likely to play key roles in the inflammatory reaction process of allergic rhinitis and polyp. Its secretion in the upper airway indicates that future studies may allow manipulation of this protein and development of novel treatments for sinonasal pathology.
Adolescent ; Adult ; Case-Control Studies ; Child ; Female ; Humans ; Male ; Middle Aged ; Nasal Mucosa ; metabolism ; pathology ; Nasal Polyps ; metabolism ; pathology ; Pulmonary Surfactant-Associated Protein A ; metabolism ; Rhinitis ; metabolism ; pathology ; Young Adult

Objective To establish an automatic review plan for coagulation tests with image recognition function,and evaluate the correctness and effectiveness of the plan.Methods Artificial intelligence software and hardware were combined to establish an image recognition system that could automatically determine the characteristics of specimens,blood volume and hematocrit.The correctness of the determination results of specimen character was compared with the visual method,the correctness of the determination results of blood volume was compared with the manual measurement method,and the correctness of hematocrit was compared with the hematology analyzer.According to the flow chart,reference interval,medical decision level,critical value range,relevant literature,work experience and historical data,the autoverification rules of coagulation tests were formulated.The autoverification rules were manually verified,and the autoverification pass rate,true positive rate,true negative rate,false positive rate,and false negative rate were calculated.The change of turnaround time in the laboratory after the implementation of the autoverification scheme was evaluated.Re-sults The accuracy rate of sample trait determination in the image recognition system was 96.72％,and the false negative rate of judging hemolytic,jaundice,and lipoid blood samples as normal samples was 0.04％.The image recognition system was compared with the blood volume data of two groups of specimens measured manually,P=0.4881.The image recognition method was not inferior to the manual measurement method.Comparing the two sets of hematocrit data from the image recognition system and the blood cell analyzer,P=0.1130,the image recognition system was not inferior to the blood cell analyzer.A total of 61 automatic re-view rules for coagulation tests had been established,including numerical abnormalities,logical abnormalities,Delta Check,sample quality abnormalities,reaction curve abnormalities,etc.The automatic review pass rate was 76.19％,true positive rate was 23.77％,true negative rate was 76.19％,false positive rate was 0.04％,and false negative rate was 0.00％.After implementing the automatic audit plan,the turnaround time of sam-ples in each quantile was shortened,with an average shortening time of 13.66 min.Conclusion The applica-tion of image recognition technology in the automatic review of coagulation tests makes the automatic review function more automated and scientific,standardizes specimen quality judgment,improves the accuracy of test results,effectively improves work efficiency and saves manpower.

Objective: To construct the third generation chimeric antigen receptor based on a novel humanized anti-HER2 H1-2 scFv, and to investigate the specific cytotoxicity of H1-2 CAR modified T lymphocytes(CAR-T) against HER2⁺ tumor cells. Method: The expression cassette of the third generation CAR gene and anti-HER2 H1-2 scFv were constructed and cloned into lentivirus transfer plasmid, and then the third generation H1-2 CAR was transduced into human T lymphocytes using lentivirus.Enzyme linked immunosorbent assay was used to detect the expression of cytokines IL2, and LDH release assay was used to detect the cytotoxic effect of the H1-2 CAR-T.Finally, NOD/SCID mice and HER2⁺ breast cancer cell line SKBR3 were used to detect the anti-tumor effect of H1-2 CAR-T in vivo. Results: The third generation H1-2 CAR was successfully constructed.H1-2 CAR-T secreted high dose of IL2 after confrontation with HER2⁺ breast cancer cells.In vitro, the cytolytic rate of H1-2 CAR-T on high expression HER2⁺ tumor cells was significantly higher than that in low expression HER2 or non-expression HER2 tumor cells. At the efficacy to target ratio of 20, the cytolytic rate of H1-2 CAR-T against breast cancer cell SK-BR-3 could reach (90.1±2.8)%, while the cytolytic rate of H1-2 CAR-T against HER2^- breast cancer cell MDA-MB-231 was only (13.5±4.7)%. In the mouse xenograft tumor model, H1-2 CAR-T cells inhibited breast cancer growth in vivo.At the end of the experiments, the average tumor weight in the H1-2 CAR-T cell treatment group was (0.7±0.1) g, the non-transfected T cell therapeutic group was (1.2±0.2) g, and the PBS group was (1.2±0.2) g. There was significant difference between the H1-2 CAR-T therapeutic group and the non-transfected T cell therapeutic group (P<0.05). However, there was no significant difference between the non-transfected T cell therapeutic group and the PBS treatment group (P>0.05). Conclusion The HER2-sepcific H1-2 CAR-T cells specifically kill HER2 positive cells, and further studies on CAR-T cells for the treatment of HER2⁺ cancers are useful.

Objective:To explore the expression and significance in regulating immune balance of programmed cell death protein 1 (PD-1) and its ligands PD-L1, PD-L2 in allergic rhinitis (AR).Methods:Eighty-two patients who received outpatient treatment due to high nasal reaction symptoms or were hospitalized due to nasal septum deviation and underwent nasal septum correction surgery in Department of Otorhinolaryngology Head and Neck Surgery of Renmin Hospital of Wuhan University from May 2018 to May 2019 were enrolled, including 42 males and 40 females, with the age ranging from 14 to 38 years old. Blood, inferior turbinate nasal mucosal tissue and relevant clinical data were collected. Patients were divided into AR group and control group due to clinical manifestation, skin prick test and detection of specific IgE (sIgE) in serum. Immunohistochemistry was used to detect the expression of PD-1 and its ligands in nasal mucosa of the two groups. Flow cytometry was used to detect the proportions of PD-1 +CD4 +T cells, PD-L1 + myeloid dendritic cells (mDCs), PD-L2 +mDCs and Th2 cells in peripheral blood of the two groups. The expression levels of total IgE, sPD-1, sPD-L1 and sPD-L2 in serum of the two groups were detected by ELISA. The measurement data of normal distribution or normal distribution after the logarithm conversion to Ln were compared by t test. Pearson correlation or Spearman correlation was used to analyze the correlation among the indicators. P<0.05 was considered statistically significant. Results:The expression of PD-1 and its ligands on the surface of immune cells in the nasal mucosa of the AR group was significantly higher than that of the control group. The ratio of PD-1 +CD4 +T cells, PD-L1 +mDCs and Th2 cells in peripheral blood of AR group was significantly higher than that of the control group ((15.24±6.45)% vs (8.71±5.33)%, (8.79±2.01)% vs (5.74±2.90)%, (7.89±1.95)% vs (2.52±1.34)%, all P<0.05). There was no significant difference in the ratio of PD-L2 +mDCs between the two groups. Correlation analysis found that the proportion of PD-1 +CD4 + T cells was positively correlated with the Visual Analogue Scale (VAS) score of AR, total IgE concentration and the serum sIgE concentration ( r value was 0.501, 0.541, 0.608, respectively, all P<0.05). The proportion of PD-L1 +mDCs was positively correlated with the VAS score of AR and the serum sIgE concentration ( r value was 0.604, 0.563, respectively, all P<0.05). The proportion of Th2 cells in peripheral blood was positively correlated with the proportion of PD-L1 +mDCs and PD-1 +CD4 +T cells ( r value was 0.538, 0.623, respectively, all P<0.05). Serum total IgE, sPD-1 and sPD-L1 in the AR group were significantly higher than those in the control group ((6.34±1.38) ng/ml vs (4.89±1.10) ng/ml, (4.40±1.01) pg/ml vs (3.79±1.21) pg/ml, (3.88±0.25) pg/ml vs (3.57±0.23) pg/ml, all P<0.05), and there was no significant difference in sPD-L2 levels between the two groups. Correlation analysis showed that sPD-L1 was positively correlated with total IgE and sIgE concentration ( r values was 0.32, 0.45, respectively, all P<0.05). Conclusions:PD-1 and PD-L1 are highly expressed on the surface of immune cells in peripheral blood and nasal mucosa of AR patients, and sPD-1 and sPD-L1 expression levels in peripheral blood of AR patients are increased. The PD-1/PD-L1 signaling pathway promote AR inflammatory response by inducing Th2 type immune response.

Objective: To facilitate using the CRISPR/Cas9 gene editing system in human liver and gallbladder cancer cells, we established Cas9 stably expressed human liver and gallbladder cancer cell lines, and validated the gene editing activity of Cas9. Methods: Human liver cancer cell lines (Huh7, PLC/PRF/5, HepG2, Hep3b, SK-HEP-1 and Li-7), human cholangiocarcinoma cells (RBE) and human gallbladder cancer cells (GBC-SD) were infected with 3 Cas9-expressing lentivirus vectors (pLv-EF1α-Cas9-Flag-Neo, pLv-EF1α-Cas9-Flag-Puro, Cas9m1.1), respectively, and Cas9 stably expressed colonies were screened and selected. We extracted the genomic DNA and protein, validated the stable expression of Cas9 by using genomic polymerase chain reaction (PCR) and western blot. Three of cell lines were further infected with Lv-EF1α-mCherry. Then mCherry positive cells were sorted by flow cytometry and infected with designed guide RNA (gRNA) vectors which targeted mCherry gene. Subsequently the gene editing activity of Cas9 was detected by genomic PCR, fluorescence microscopic observation and flow cytometry analysis. Results: One hundred Cas9-expressing human liver and gallbladder cancer cell lines were selected. Among them, 35 cell lines expressed Cas9-Neo, 25 expressed Cas9-puro, and 40 expressed mutant Cas9 (mCas9). We also established 3 cell lines with stable expression of mCherry (Huh7-mCas9-M, PLC/PRF/5-Cas9-M and SK-HEP-1-Cas9-M). The results of genomic PCR and sequencing showed that by lentiviral infection with 2 types of designed gRNA, the long fragment deletion of mCherry gene was found in these 3 cell lines. Moreover, mCherry^-EGFP⁺ cells infected with 2 types of gRNA were observed by fluorescence microscope. The results of flow cytometry showed that mCherry^-EGFP⁺ cells accounted from 0.3% to 93.6%. Conclusion We successfully establish 100 human liver and gallbladder cancer cell lines with stable expression of Cas9 protein and validate their activities of gene editing.