To investigate the correlation between p16 gene mutation and laryngeal cancer biological behavior as well as its prognosis in laryngeal cancer.Method:24 speciments of primary laryngeal cancer and 10 speciments with benign lesion in larynx were examined for mutations in exon2 of p16 by using PCR-SSCP silver stainning technique.Result:Mutations frequency of laryngeal cancer was62.5% (15/24).Nothing was found in 10 cases with laryngeal benign lesion.Conclusion:There is a strong correlation between p16 gene mutation and the biological behavior of chinese laryngeal cancer, such as histologic differentiation, invasion stage, and regional lymph nodes metastasis(P≤0.05).PCR-SSCP silver tainning technique is one of the most sensitive and simplest measure for detecting genetic mutation.It is worth using in clinical laboratory because of its readiness, repetition and lower cost.

Objective To investigate the effect of capsaicin and cisplatin on the proliferation and immunogenic cell death of human osteosarcoma cells.Methods MTT assay was used to examine the growth inhibiting effects of capsacin and cisplation on MG-63 cells;Mitochondrial membrane potential (MMP) was used to investigate the apoptosis;flow cytometry was used to detect the expression of calreticulin (CRT) on the cell membrane,fluorescein enzymatic method was used to detect the release of ATP,ELISA was used to detect the secretion of high mobility group B1 (HMGB1).Results Capsaicin and cisplatin can inhibit MG-63 cells proliferation in the dose-dependent manner and induce MG-63 apoptosis(P＜0.01).Only capsaicin can induce translocation of CRT from endoplasmic reticulum to the cell surface and release of extracellular ATP and HMGB1 (P＜0.01).Conclusions Capsaicin can induce human osteosarcoma cells apoptosis and immunogenic cell death.

Objective:To investigate the protective role of heme oxygenase-1 and its reaction product,carbon monoxide against acute liver injury induced by carbon tetrachloride in rats.Methods: Thirty male Sprague-Dawley rats were randomly divided into six groups with five in each.The control group received a single dose of corn oil injection.Carbon tetrachloride was injected intraperitoneally(i.p) to establish acute liver injury models in rats.Hemin(50 ?mol/kg) was administered i.p.12 hours before CCl_4 treatment,with an aim to induce HO-1 protein expression in the liver of rats.Carbon monoxide was injected i.p.12 hours prior to CCl_4 injection,resulting in about 8%-12% carboxyhemoglobin concentration in vivo.The expression of HO-1 in the liver of hemin-treated rats was determined by western blot method at different time points.At 24 h after carbon tetrachloride administration,all rats were sacrificed to collect blood samples for the examination of ALT,AST levels and to remove liver tissues for analysis of MDA concentration,SOD activity and caspase-3 activity as well as TNF-a contents.In addition,histopathological changes were investigated and hepatocyte apoptosis was detected by TUNEL method.Results: The administration of carbon tetrachloride to rats caused a marked hepatic damage,characterized by significant elevation of serum ALT,AST levels(2 136.3?163.4 U,1 422.7?221.7 U) and liver MDA con-tent(5.28?0.93 ?mol/g),caspase-3 activitiy(optical density value(4.69)?1.02) and TNF-? level(256.3?27.3 ng/L) combined with a remarkable reduction in liver SOD activity(45.9?14.8 U/mg) as compared with the control rats.Histopathological observations revealed severe damage in the liver and prominent hepatocyte apoptosis took place in CCl_4treated rats.However,pretreatment with hemin could induce high expression of HO-1 protein and exert potent protective effects against liver injury,as demonstrated by a significant decrease in ALT,AST levels(287.1?24.3 U,246.2?21.7 U) and MDA concentration(3.27?1.34 ?mol/g),reduction in caspase-3 activity(optical density value 2.49?1.47) and TNF-? level(132.6?19.5 ng/L),as compared with the CCl_4-treated rats.Moreover,hepatocyte apoptosis and liver injury were both attenuated remarkably in the liver of rats pretreated with hemin.In contrast to hemin administration,single injection of exogenous CO produced the same protective effects,as indicated by the remarkable reduction of ALT,AST levels and caspase-3 activity and TNF-a levels.Conclusion: The above results suggest that HO-1/CO system has a potent protective effect on acute liver injury induced by carbon tetrachloride in rats.Induction of HO-1 expression and low concentration of CO can inhibit the progress of hepatic damage,which might be due to the alleviation of lipid peroxidation and reduction of caspase-3 activity or inhibition of TNF-? level.

OBJECTIVE:To evaluate the quality of levofloxacin tablets produced by 6 different factories.METHODS:To investigate the quality of 17 batches of levofloxacin tablets produced by 6 factories according to related standards and to evaluate the drug dissolubilities using paddle method.RESULTS:17 products from six factories were all proved qualified.Significant differences of dissolution parameters and treatment costs were found among the different products.CONCLUSION:There are difference in quality of levofloxacin tablets.The tablets produced by the factory holding the patent right are more expensive,however the qualiity is more stable.

BACKGROUND: The use of allograft tendon promotes the development of anterior cruciate ligament reconstruction, which is of great importance in the relevant basic research and clinical practice. OBJECTIVE: To summarize the current application and progress of allograft tendon in the anterior cruciate ligament reconstruction. METHODS: We searched relevant articles about the use of allograft tendon in PubMed and Embase published from January 2012 to February 2018 using the keywords of "allograft tends OR allografts, intra-articular knee ligament OR anterior cruciate ligament OR ACL". RESULTS AND CONCLUSION: China's use of allogeneic tendon in the reconstruction of the anterior cruciate ligament is still in the developmental stage. The current pros and cons of using allografts or autografts in anterior cruciate ligament reconstruction are inconclusive. Concerns about the choice of allografts are mainly due to possible infections and slow healing. A huge difference exists in different trials concerning the effects of allografts versus autografts. The conclusions are therefore roughly as follows: there is no difference in knee function after implantation of the two grafts, or the knee joint function is better after autograft. Studies have shown that allografts may increase the risk of secondary surgery. Therefore, the use of allograft tendon faces many clinical challenges in the anterior cruciate ligament reconstruction.

Objective To investigate the efficacy and pregnancy outcomes of women receiving double-catheter epidural block in labor analgesia, and compare the results with single-catheter epidural block.Methods A double-blind clinical trial was conducted on 206 full-term singleton primiparas, aged 25-35 and at the 37 -42 weeks of gestation who delivered at the Department of Obstetrics, Qingdao Municipal Hospital from August 2006 to December 2008, which were randomly divided into two groups:double-catheter epidural block ( group D, n = 103) and single-catheter epidural-block ( group S, n = 103 ).Women in group D were given mixture of 0.1% repivacaine hydrochloride and 0.5 mg/L sufentinil 4 -6 ml as initial dose.Patient control epidural analgesia pump (PCEA) was connected with the upper catheter after 45 minutes.A bolus dose of 4 -6 ml analgesia mixture was infused according to the condition through the lower catheter.Women in group S received analgesia mixture 10 - 15 ml as initial dose and PCEA pump was connected after 45 minutes.Oxytocin was infused in both groups according to uterine contraction after 30 minutes.The following indexes was observed: ( 1 ) visual analogue scales (VAS); (2) modified Bromage Scores;(3) the total dose of analgesia mixture, the percentage of oxytocin infusion, duration of labor and duration of the second stage of labor; (4) fetal birth weight and Apgar scores( 1,5 minutes); (5) mode of delivery; (6) the concentration of plasma cortisol and angiotension Ⅱ at the beginning of regular uterine contraction and at the time when cervical dilated to 4 cm and 10 cm and fetal disengagement; (7)anesthesia-related complications.Results ( 1 )The neonatal birth weight and Apgar scores ( 1,5 minutes)of group D were (3456 ±468)g, 9.8 ±0.6 and 9.9 ±0.7, respectively, while(3399 ±569) g, 9.8 ±0.5 and 9.9 ±0.7 in group S( P ＞0.05).No motor function block was reported in any group and the modified Bromage score was zero.(2) The total dose of analgesia mixture in group D was similar to that in group S [(57 ±9) ml vs.(58 ±11) ml, P＞0.05].However, the percentage of women received oxytocin in group D was smaller [59.2% (61/103) vs.81.6% (84/103), P ＜ 0.01], and the total time of labor and the duration of second stage of labor in group D were shorter[(532 ± 140) minutes vs.(608 ± 150) minutes;(46 ± 31 ) minutes vs.(60 ± 34) minutes, P ＜ 0.05].(3) There were no significant differences in VAS at 30 minutes after initial dose and in the first stage of labor between group D and S ( 1.2 ± 1.1 vs 1.2 ± 1.1,1.1 ± 1.1 vs.1.2 ± 1.0, P＞0.05).VAS at the second stage of labor stage was lower in group D than in group S ( 1.2 ± 1.1 vs.4.5 ± 2.2, P ＜ 0.01 ).(4) The rate of cesarean section, instrumental delivery and episiotomy in group D were lower than in group S (7.8% vs.17.5%, 7.8% vs.15.5%, 10.7% vs.18.4%, P ＜ 0.05).The incidence of fetal distress and meconium-stained amniotic fluid as the indication of cesarean section were similar between the two groups (P ＞ 0.05 ).Lower incidence of fetal malpresentation and arrested second stage of labor were shown in group D than in group S (2.9% vs.9.7%, 1.0% vs.5.8%, P ＜ 0.05 ).(5) The concentration of plasma cortisol and angiotension Ⅱ were lower in group D than in group S [(86 ±25) ng/L vs.( 100 ±20) ng/L, (278 ±53) nmol/L vs.(311 ±53)nmol/L, P＜0.05] only at the end of second stage of labor, but not at any other times(P ＞0.05).(6) No serious anesthesia-related complications were reported in any groups.Some light backache around the puncture point were complained by 29.1% (30/103) of the women in group D and 31.1% (32/103) in group S(P ＞0.05).Conclusion Double-catheter epidural block can provide better analgesia effect during labor than single-catheter epidural block, without any adverse influence on delivery outcomes.

Objective:To evaluate the value of cerebrospinal fluid (CSF) nerve injury-related proteins levels in predicting postoperative delirium (POD) in patients.Methods:A total of 1 000 patients of both sexes, aged 40-90 yr, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, with Mini-Mental State Examination score>24 at 1 day before operation, undergoing elective knee/hip arthroplasty under spinal-epidural anesthesia , were enrolled in this study.Cubital venous blood samples were drawn before anesthesia for detection of the concentrations of plasma total cholesterol, high-density lipoprotein, low-density lipoprotein and triglyceride.CSF 2ml was extracted after successful spinal-epidural anesthesia puncture for measurement of concentrations of α-synuclein (α-syn), β-amyloid protein 1-40 (Aβ 1-40), Aβ 1-42, total-Tau (t-Tau), phosphorylated Tau (p-Tau), progranulin (PGRN) and soluble myeloid cell triggering receptor 2 (sTREM2) (by enzyme-linked immunosorbent assay). The Confusion Assessment Method was used at 1, 3 and 7 days after surgery to evaluate the occurrence of POD.The patients were divided into POD group and non-POD group according to whether POD occurred after operation.Logistic regression analysis was used to analyze the variables of which P values were less than 0.05 to analyze the risk factors for POD.The receiver operating characteristic (ROC) curve was drawn and area (AUC) under the curve was calculated to evaluate the accuracy of the related risk factors in predicting POD. Results:A total of 964 patients were enrolled in the study, and 108 patients were diagnosed with POD, and the incidence was 11.2%.The results of logistic regression analysis found that age and and increased α-syn in CSF concentration were risk factors for POD, and decreased PGRN in CSF concentration and Aβ 1-42/p-Tau in CSF were the protective factors for POD ( P<0.05). ROC curve analysis showed that α-syn (AUC 0.69, 95% confidence interval (CI) 0.634-0.748, sensitivity 57.41%, specificity 82.10%, Youden Index 0.3951), PGRN in CSF concentration (AUC 0.695, 95%CI 0.637-0.750, sensitivity 59.26%, specificity 80.86%, Youden Index 0.4012) and Aβ 1-42/p-Tau in CSF (AUC 0.635, 95%CI 0.574-0.692, sensitivity 93.52%, specificity 30.25%, Youden Index 0.2377) could predict the occurrence of POD. Conclusion:PGRN, α-syn concentration and Aβ 1-42/p-Tau in CSF can predict the occurrence of POD in patients.

Objective:To evaluate the relationship between preoperative cerebrospinal fluid/serum albumin ratio (Q-alb) and postoperative delirium (POD) in patients undergoing neuraxial anesthesia.Methods:The patients, aged 40-90 yr, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, underwent total knee/hip replacement under combined spinal-epidural block in our hospital from January 2018 to December 2020, were collected.After admission to the operating room, venous blood and cerebrospinal fluid samples were collected for determination of cerebrospinal fluid albumin, β-amyloid (Aβ) 1-42, Aβ 1-40, total tau protein (t-Tau), phosphorylated tau protein (p-Tau) and serum albumin levels (by enzyme-linked immunosorbent assay) and for calculation of Q-alb.When Q-alb was more than 10.2, the patient was considered to have blood-brain barrier disruption.Mini-Mental State Examination scale was used to evaluate the cognitive level on 1 day before surgery. The development of POD was evaluated using Confusion Assessment Method Chinese Reversion and Memorial Delirium Assessment Scale at 1-7 days after surgery.The patients were divided into POD group (P group) and non-POD (NP group) according to whether POD occurred.The receiver operating characteristic (ROC) curve was used to analyze the accuracy of Q-alb in predicting POD. Results:There were 49 cases in each group.Compared with group NP, concentrations of Aβ 1-42 and Aβ 1-40 were significantly decreased, concentrations of t-Tau and p-Tau albumin were increased, the ratio of Q-alb and blood-brain barrier disruption was increased in group P ( P<0.05). Before and after adjusting for confounding factors, Q-alb, cerebrospinal fluid Aβ 1-42, Aβ 1-40, t-Tau and p-Tau levels were risk factors for POD ( P<0.05). There was a positive linear regression relationship between Q-alb and levels of t-Tau and p-Tauin cerebrospinal fluid (t-Tau: β=0.587, P<0.001; p-Tau: β=0.427, P<0.001), and there was a negative linear regression relationship between Q-alb and levels of Aβ 1-42 and Aβ 1-40 in cerebrospinal fluid (Aβ 1-42: β=-0.762, P<0.001; Aβ 1-40: β=-0.531, P<0.001). There was no linear regression relationship between Q-alb and level of p-Tau in group P ( P=0.121). There was no linear regression relationship between Q-alb and level of Aβ 1-40 in group NP ( P=0.467). The results of ROC curve analysis showed that the area under the curve for Q-alb in predicting POD (95% confidence interval) was 0.827 (0.738-0.896). Conclusion:Preoperative higher Q-alb is the risk factor for POD in patients undergoing neuraxial anesthesia, and is more accurate in predicting POD.

Objective:To investigate the association of R-loop binding proteins with prognosis and chemotherapy efficacy in lung adenocarcinoma.Methods:The data related to R-loop regulatory genes were obtained from literature of R-loop proteomics and relevant databases.We used 403 cases of lung adenocarcinoma in the Cancer Genome Atlas as training set,and two datasets GSE14814 and GSE31210 in Gene Expression Omnibus as validation sets.The weighted gene co-expression network analysis(WGCNA)was employed to identify R-loop genes with a significant impact on the clinical phenotype of lung adenocarcinoma.Least absolute shrinkage and selection operator(LASSO)regression analysis was utilized to eliminate genes exhibiting multicollinearity.A multivariate Cox regression analysis was employed to scrutinize clinical variables and R-loop characteristic genes that exert independent prognostic effects on patient survival.Subsequently,a risk score model was constructed.The predictive capacity of this model for the prognosis of patients was analyzed and validated.Additionally,the performance of risk model on the anti-tumor drug sensitivity was assessed.The mutations of R-loop genes were analyzed by maftools.The effect of PLEC expression on anti-tumor drug sensitivity was tested on non-small cell lung adenocarcinoma H1299 and A549 cells in vitro.Results:A collection of 1551 R-loop genes were obtained,and 78 genes exhibited significant effects on the clinical phenotype shown on WGCNA.The LASSO regression analysis retained fourteen R-loop genes.A multivariate Cox regression analysis further identified three R-loop genes(HEXIM1,GLI2,PLEC)and a clinical variable(tumor grading)that were associated with patient prognosis.Risk prediction model was established according to the regression coefficients of each parameter.Kaplan-Meier survival analysis showed that the prognosis of high-risk group was significantly worse than that of low-risk group(P<0.01).The time-dependent ROC curve showed that the risk model had good predictive ability in both training and validation sets.Predictive analyses of anti-neoplastic drug sensitivity indicated a diminished responsiveness to both chemotherapy and targeted treatment drugs among high-risk patients.The expression of PLEC was strongly correlated with sensitivity to gefitinib,a classical EGFR inhibitor.Conclusion:R-loop binding proteins have been identified as significant determinants in the prognosis and therapeutic strategies for lung adenocarcinoma,which indicates that therapeutic interventions targeting these specific R-loop binding proteins might contribute to a better survival of the patients.

Objective:To rapidly evaluate the level of healthcare resource demand for laboratory testing and prevention and control of corona virus disease 2019 (COVID-19) in different epidemic situation, and prepare for the capacity planning, stockpile distribution, and funding raising for infectious disease epidemic response.Methods:An susceptible, exposed, infectious, removed infectious disease dynamics model with confirmed asymptomatic infection cases and symptomatic hospitalized patients was introduced to simulate different COVID-19 epidemic situation and predict the numbers of hospitalized or isolated patients, and based on the current COVID-19 prevention and control measures in China, the demands of resources for laboratory testing and prevention and control of COVID-19 were evaluated.Results:When community or local transmission or outbreaks occur and total population nucleic acid testing is implemented, the need for human resources is 3.3-89.1 times higher than the reserved, and the current resources of medical personal protective equipment and instruments can meet the need. The surge in asymptomatic infections can also increase the human resource demand for laboratory testing and pose challenge to the prevention and control of the disease. When vaccine protection coverage reach ≥50%, appropriate adjustment of the prevention and control measures can reduce the need for laboratory and human resources.Conclusions:There is a great need in our country to reserve the human resources for laboratory testing and disease prevention and control for the response of the possible epidemic of COVID-19. Challenges to human resources resulted from total population nucleic acid testing and its necessity need to be considered. Conducting non-pharmaceutical interventions and encouraging more people to be vaccinated can mitigate the shock on healthcare resource demand in COVID-19 prevention and control.