Objective:To study the value of cell paraffin block immunohistochemistry and pleural fluid Crk like protein (CRKL) and macrophage inhibitory cytokine-1 (MIC-1) in the diagnosis of malignant pleural effusion.Methods:A total of 98 patients with pleural effusion treated in Shantou Central Hospital from February 2020 to February 2021 were retrospectively selected as the research objects, including 58 benign cases and 40 malignant cases. The levels of CRKL and MIC-1 in pleural effusion were detected by enzyme-linked immunosorbent assay. The pleural effusion was analyzed by cell paraffin block immunohistochemistry. The levels of various indexes in benign group and malignant group were compared. The diagnostic value of cell paraffin block immunohistochemistry and pleural effusion CRKL and MIC-1 for benign and malignant pleural effusion was analyzed by receiver operating characteristic (ROC) curve.Results:With pathological results as the gold standard, 54 cases of benign and 44 cases of malignant were diagnosed by cell paraffin block immunohistochemistry. The diagnostic accuracy was 75.5% (74/98) , and the sensitivity and specificity were 75.0% (30/40) and 75.9% (44/58) respectively. The levels of pleural effusion CRKL [2.84 (2.17, 3.98) ng/ml vs. 1.88 (0.94, 2.62) ng/ml], MIC-1 [2.28 (1.67, 2.98) ng/ml vs. 1.76 (1.22, 2.32) ] ng/ml] in the malignant group were higher than those in the benign group, with statistically significant differences ( Z=-4.57, P<0.001; Z=-3.09, P<0.001) . The optimal critical value of CRKL in pleural effusion for the diagnosis of malignant pleural effusion was 2.33 ng/ml, the area under the curve (AUC) was 0.76 (95% CI: 0.66-0.85) , and the sensitivity and specificity were 67.5% (27 /40) , 74.1% (43/58) . The optimal critical value of MIC-1 in pleural effusion for the diagnosis of malignant pleural effusion was 2.10 ng/ml, the AUC was 0.74 (95% CI: 0.64-0.85) , and the sensitivity and specificity were 60.0% (24/40) , 82.8% (48/58) . The AUC of MIC-1 and CRKL in pleural effusion combined with cell paraffin block immunohistochemistry for the diagnosis of malignant pleural effusion was 0.83 (95% CI: 0.75-0.91) , and the sensitivity and specificity were 85.0% (34/40) and 70.7% (41/58) . The sensitivity and AUC of combined diagnosis were significantly higher than those of CRKL and MIC-1 alone (sensitivity: χ2=4.26, P=0.046; χ2=6.27, P=0.012; AUC: Z=3.53, P<0.001; Z=4.14, P<0.001) . Conclusion:CRKL and MIC-1 in pleural effusion of patients with malignant pleural effusion are highly expressed, which can be used as indicators for the diagnosis of malignant pleural effusion. Detection combined with cell paraffin block immunohistochemistry can improve the diagnostic value of malignant pleural effusion.

OBJECTIVE To isolate and ide ntify the chemical constituents of the root of Ardisia virens and preliminarily evaluate the in vitro anti-inflammatory activity of the compounds. METHODS The ethyl acetate extraction part from 70％ ethanol extract of the root of A. virens were separated and purified by silica gel column chromatography ，ODS column chromatography ， etc. The structures of the compounds were identified according to physical and chemical properties and spectral data. The inflammation model of RAW 264.7 cells was induced by lipopolysaccharide ，and anti-inflammatory activity of the compound was investigated by MTT assay. RESULTS A total of 11 compounds were isolated from the ethyl acetate extraction part ，and were identified as cyclamiretin A （1），α-spinasterol （2），（3S，5R，6S，7E）-3，5，6-trihydroxy-7-megastigmen-9-one （3）， （+）-angelicoidenol（4），octadeca-dienoic acid- 2，3-dihydroxypropyl ester （5），α-linolenic acid （6），glycerol monooleate （7），5， 5′-（4，7-hexadecadlene-1，16-diyl）bisresorcinol（8），1-（3，5-dihydroxyphenyl）heptan-1-one（9），5-heptylresorcinol and （10） 5-n-nonylresorcinol（11）. The in vitro anti-inflammatory results showed that 80，40，20，10，5 μg/mL of compounds 2，8，9 and 10 could reduce the cell survival rate in different degrees. CONCLUSIONS Compounds 1-11 are isolated from this plant for the first time，and compound 8 is a new natural product. Compound 2，8，9 and 10 show certain anti-inflammatory activity in vitro .

Objective To analyze the infiltration of inflammatory cells under the mucosa of female cystitis glandularis and the different inflammatory infiltration in different clinical pathological types of cystitis glandularis.Methods Immunohistochemical method was used to detect the bladder mucosal tissue samples of 10 female patients confirmed cystitis glandularis admitted from June 2016 to October 2016.The results of immunohistochemical staining were collected and statistically analyzed by the automatic microscopy and image analysis system.In addition,the clinical data and tissue sample of 49 cases of cystitis glandularis treated from December 2006 to August 2017 were collected.Age of 49 patients was (34.4 ±7.5) years old and BMI was (21.9 ± 4.2) kg/m2.There were 19 cases of hypertension and 18 cases of diabetes.According to the cystoscopic manifestations,follicular edema type,papilloma type,and intestinal adenomatosis type were defined as high risk.Chronic inflammatory type and mucosa unchanged type were defined as low risk.Immunohistochemical staining was used to detect tissue samples,to compare the general data of different types of cystitis glandularis and the degree of infiltration of bladder mucosal inflammatory cells.Results T lymphocytes were highly expressed in 10 patients,and B lymphocytes and plasma cells were not expressed or extremely low (P ＜ 0.01).Of the 49 patients,29 were high risk type cystitis glandularis (follicular edema type,papilloma type,and intestinal adenomatosis type),and 21 were low risk type (chronic inflammatory type and mucosa unchanged type).The age of the high-risk group was (34.4 ± 7.5) years old with BMI of (21.9 ±4.2) kg/m2,8 cases of hypertension and 8 cases of diabetes.The age of the low-risk group was (38.2 ±8.5) years old with BMI of (20.8 ±4.0) kg/m2,11 cases of hypertension and 10 cases of diabetes.There was no statistically significant difference between two groups (P ＞ 0.05).The OABSS of high-risk group(10.4 ± 2.6) was significantly higher than that of low-risk group (7.1 ± 2.1,P ＜ 0.01).QOL of high-risk group (4.9 ± 0.9) was significantly higher than that of low-risk group (4.1 ± 0.8,P ＜ 0.01).Qmax of high-risk group was (11.4 ± 3.6) ml/s,significantly lower than that of low-risk group[(15.8 ±3.8) ml/s,P ＜0.01].The positive number of T lymphocytes of high-risk group was (173.5 ± 26.8),which was significantly higher than that of low-risk group(119.5 ± 21.2,P ＜ 0.01).Conclusions T lymphocytes infiltration is the major phenomenon in bladder submucosa of female patients with cystitis glandularis.The inflammatory infiltration by T lymphocytes could be associated with patient's symptom and bladder's pathological changes.

Objective: To investigate the relationships between the expression of programmed death 1 (PD-1) and the epidermal growth factor receptor (EGFR) gene mutations in non-small cell lung cancer (NSCLC). The study also attempted to investigate the clinicopathological features and prognosis in NSCLC patients. Methods: The expression of PD-1 protein in 88 cases of NSCLC tumor tissues and adjacent tissues was detected by immunohistochemistry. The mutations of EGFR in NSCLC were detected by Polymerase Chain Reaction-Amplification Refractory Mutation System(PCR-ARMS) method. The expression of PD-1 and patients′ clinical characteristics and prognosis were analyzed. Results: PD-1 was positive in 63.6%(56/88) NSCLC tumor tissues, which was significantly higher than that in adjacent normal tissues (21.6%, 19/88) (P<0.05). EGFR gene mutations were found in 43 cases (48.9%), in which 30 cases (69.8%)were PD-1 positive expression. 45 cases had the wild types of EGFR gene, in which 26 cases (57.8%) were PD-1 positive. There were 24 cases of 19Del EGFR mutations, including 20 cases (83.3%) of PD-1 positive expression. 19 patients had 21L858 EGFR mutations, including 10 cases (52.6%) of PD-1 positive expression. The expression of PD-1 in NSCLC was related to patients′ smoking status, lymph node metastasis and EGFR gene mutations (P<0.05). The median progression-free survival time of patients with PD-1 positive and negative expression was 7.03 and 18.66 months, respectively (P=0.007). In patients with wild-type EGFR gene, the median progression-free survival time of PD-1 positive and negative expression was 25.21 and 38.24 months, respectively. The difference was statistically significant (P=0.024). The median progression-free survival time in 43 cases of EGFR mutant patients with PD-1 positive and negative expression was 21.23 and 31.44 months. The difference was not statistically significant (P=0.128). Conclusions PD-1 expresses in both EGFR mutant and wild-type NSCLC, and its expression levelis different with various EGFR mutations. The expression of PD-1 in NSCLC is related to the prognosis of patients, and the prognosis of patients with positive PD-1 expression was poor.

Objective To report the clinical effects of a 3D-printed metal drilling template used in surgery for complex upper cervical disorders.Methods Between August 2015 and June 2017,5 patients with upper cervical disorder complicated with complex cranial cervical junction deformity underwent instrumentation and fusion.They were 4 males and one female,aged from 3 to 36 years.All the patients underwent thin slice CT scan before operation.The CT image data were imported into a computer workstation for three dimensional reconstruction of individualized models of the upper cervical spine on which individualized drilling templates were designed.Guided by each of the individualized metal drilling templates manufactured by 3D printing,surgery via the posterior approach was performed in 2 cases and surgery via the combined anterior and posterior approach in 3 cases.After surgery,the screw trajectory was evaluated by CT scan.Results Surgery succeeded in all the 5 patients.A total of 5 pedicle screws and 5 laminar screws were drilled.Postoperative CT scan showed accurate placement of all the 10 screws.The patients recovered with no neurovascular complications.Conclusion Since an individualized 3D-printed metal drilling template can improve the accuracy and safety of screw placement,it can reduce surgical risks and enhance surgical success in the surgery for complex upper cervical disorders.

BACKGROUND:Artificial knee joint replacement is the main surgical method for the treatment of severe knee joint deformation, but due to the limited existing knee prosthesis type. The stature of Fujian population in southeast China was shorter than that of the north. The size of artificial knee joint usual y did not fit for patients in clinic. The substitution effect was poor.
OBJECTIVE:To measure the geometric parameters of proximal surface and cross-section of adult female tibia in southeastern area of Fujian Province of China, and provide basic data for design and replacement of artificial knee joint in females.
METHODS:We chose 85 cases of dry female skeleton in southeastern area of Fujian Province of China, measured tibial plateau width, medial tibial plateau width and anteroposterior diameter, lateral tibial plateau width and anteroposterior diameter. We col ected clinical normal proximal tibial CT data of 36 patients (38-65 years old), with scan range of 30 cm (intercondylar eminence 30 cm) and layer thickness of 0.625 mm. Mimics 16.0 three-dimensional reconstruction software was used to measure above indexes and cut the reconstructed models with thickness of 3 mm, total y 10 layers. We divided its widest distance into four equal parts, and measured tibial anteroposterior diameter on three points. Statistical software was used to analyze the linear relationship between the data.
RESULTS AND CONCLUSION:The experiment has successful y obtained geometric parameters of tibial specimens of adult females in southeastern area of Fujian province of China. Tibia platform inside and outside diameters were greater than the width inside and outside;inside and outside diameter and inside and outside width were similar. Ten segment planes of tibia, inside and outside diameters were always less than middle anteroposterior diameter. Three groups of data increased firstly, and then reduced. There were positive correlations between tibia length and width of the medial tibia platform, the lateral tibia platform width and anteroposterior diameter, the medial tibia platform width and anteroposterior diameter, and the lateral tibia platform width and anteroposterior diameter (P<0.05). These results suggested that relevant parameters of tibial plateau adult women in southeastern area of Fujian province of China showed some regular changes. Our results may provide quantitative reference data for the design and operation of artificial tibial plateau.

Eosine Yellowish-(YS) ; Frozen Sections ; Hematoxylin ; Humans ; Staining and Labeling

Objective To understand and analysis of the pathogens distribution and drug resistance of nosocomial infection in children′s hospital,so as to provide reliable scientific basis for the prevention and control of hospital infection.Methods 396 cases of upper respiratory tract specimens were collected from pediatric patients with nosocomial infection.These specimens were detected by sputum specimens conventional methods of microorganism cultivation,and K-B method was used to determine the bacteria sensi-tivities to clinical common drug.Results There were 225 cases of specimens were pathogen positive among all the 396 specimens, and 234 strains of bacteria were isolated in all.The positive isolated rate was 56.8%(225/396).Among the 234 isolated strains, Gram negative bacteria accounted for 72.6%(170/234),and Klebsiella occupied the first place[49.4%(84/170)].Gram positive bacteria accounted for 23.5%(55/234),and Staphylococcus had the highest isolated rate in Gram positive bacteria[58.2% (32/55)].In all the 9 kinds of clinical common antimicrobial agents,imipenem had high drug sensitivity to the 234 isolated strains,and the aminoglycosides came next.Conclusion It is necessary for the pediatric patients with nosocomial infection to collect upper re-spiratory tract specimens for bacteriologic studies and drug sensitivity tests.

Objective To investigate the effect of herpes simplex virus thymidine kinase/ganciclovir (HSV-TK/GGV)system driven by human alpha fetoprotein(AFP)enhancer on hepatocellular carcinoma(HCC)cells in vitro and in vivo.Methods HCC-specific eukarotypic expression vector carrying suicide gene driven by AFP enhancer(pAFP-cDNA3.1-TK)was constructed.The plasmid was trasfected to AFP-positive HepG2 cells and AFP-negative SMMC7721 cells by liposomes.Protein and mRNA expressions of TK were detected by RT-PCR or Western blot.The survival rates of HCC cells were detected by methyl thiazolyl tetrazolium assay.The effects of GGV on the in vitro proliferation,survival and apoptosis of HCC cells were observed,and the inhibitive effect of GGV on the survival of HCC cells in vivo was also detected.All data were analyzed by using the t test.Results The pAFP-cDNA3.1-TK was successfully constructed and transfected to the HCC cells.The protein and mRNA expressions of TK were detected in AFP-positive HepG2 cells.GGV dose-and time-dependently inhibited the growth and induced the apoptosis of HepG2 cells in vitro,but it had no effect on SMMC7721 cells.No protein or mRNA expression of TK was detected in the SMMC7721 cells.There was a significant difference on the inhibitory effects of GGV on HepG2 cells and SMMC7721 cells(t =2.58,2.73,3.12,P ＜0.05).GGV specifically inhibited the growth of AFP-positive HepG2 cells,and the inhibition rate was 46%;the growth of AFP-negative SMMC7721 cells was not influenced by GGV.There was a significant difference in the inhibitive effect of GGV on the growth of HepG2 cells and SMMC7721 cells(t = 3.36,P ＜ 0.05).Conclusion HSV-TK/GGV systemdriven by human APF enhancer kills APF-positive HCC cells and inhibits the growth of HCC cells.

Objective To investigate the effect of cerebral ischemia on functional parameters of affected arteries and probe into the possible pathogenesis of ischemia-reperfusion injury.Methods Intraluminal suture ischemic model was used by occlusion of left middle cerebral artery in rats.Two hours later,the middle cerebral artery segments were isolated from both ischemia and control groups for measurement of changes in vessel diameter induced by increasing pressure and vasoactive compounds.And then,distensibility,myogenic tone,reactivity to 5-HT and ACh were calculated and compared between groups.Results In lower pressure range,ischemic vessels showed an increased myogenic tone(at 40 mm Hg,1 mm Hg=0.133 kPa,19.3%±0.4% vs 10.0%±0.2%,t=20.568,P=0.000)and decreased diameter.In higher pressure range,ischemic vessels showed an increased diameter.distensibility and decreased myogenic tone(at 120 mm Hg,12.0%±0.2% vs 21.8%±0.4%,t=-23.575,P=0.000).In normal pressure range,myogenic tone was not altered after ischemia. Both groups constricted to 5-HT and dilated to ACh,however,the response was significantly diminished after ischemla.Conclusion These findings demonstrate that contractile and diastolic function of affected artery was impaired after ischemia,a result that may contribute to ischemia-reperfusion injury by losing upstream cerebrovascular resistance and increasing perfusion on the microcirculation.