Objective: To examine expression patterns of focal adhesion kinase (FAK) and its activated form, phosphorylated FAK (pFAK),in human osteosarcoma and to investigate the correlation of FAK expression with clinicopathological parameters and prognosis. Functional consequence of manipulating FAK protein levels was also investigated in human osteosarcoma cell lines. Methods: Immunohistochemical staining was used to detect FAK and pFAK levels in pathologically archived materials from 113 patients with primary osteosarcoma. Kaplan-Meier survival and Cox regression analyses were used to evaluate prognoses. The role of FAK in cytological behavior of MG63 and 143B human osteosarcoma cell lines was studied via the FAK protein knockdown with siRNA. Cell proliferation, migration, invasiveness, and apoptosis were assessed using cell counting kit-8, Transwell, and Annexin V/PI staining methods. Results: Both FAK and pFAK were overexpressed in osteosarcoma patients. Tumor cells exhibited cytoplasmicity and occasional membranous immunoreactivity for FAK. A total of 42 cases (37.17%) mainly showed expressed pFAK in cytoplasm of osteosarcoma cells. No overexpression staining of anti-FAK and anti-pFAK antibodies was observed in normal cancellous bone tissues or negative controls. Significant differences were observed in overall survival between FAK-/pFAK- and FAK+/pFAK- groups (P=0.016), FAK+/pFAK- and FAK+/pFAK+ groups (P=0.012), and FAK-/pFAK- and FAK+/pFAK+ groups (P<0.001). All groups showed similar metastasis-free survival. Cox proportional hazard analysis showed that FAK expression profile is an independent indicator of both overall andmetastasis-free survival. siRNA-based knockdown of FAK significantly reducedmigration and invasion of MG63 and 143B cells and affected proliferation and apoptosis in osteosarcoma cells. Conclusion: Osteosarcoma malignancies in vitro and in vivo were correlated with overexpression and phosphorylation of FAK. These findings suggest that FAK plays an important biological role in osteosarcoma carcinogenesis. This study provides a better understanding of diagnostic and prognostic relevance of FAK overexpression and phosphorylation in osteosarcoma patients. Therefore, FAK and pFAK can be used as independent predictors of overall and metastasis-free survival in osteosarcoma patients.

Objective To investigate the expression and variation of MIP 1β,MIP-2,and IL-12p70 in mice with bloodstream infection caused by 4 kinds of bacteria.Methods CD-1 (ICR) mouse models of bloodstream infection with Staphylococcus aureus (S.aureus),Enterococcus f aecalis (E.f aecalis),Escherichia coli (E.coli),and K lebsiella pneumoniae (K.pneumoniae) were established.After mice in each trial group and PBS control group were infected by bacteria for 0.5h,1h,3h,6h,12h,24h,and 48h,concentrations of MIP-1β,MIP-2,and IL-12p70 were detected by Luminex liquid suspension chip system.Results Concentrations of MIP-1β increased significantly 1h after bacteria was in blood,S.aureus,E.faecalis,E.coli,K.pneumoniae,and control groups were (134.5 ± 18.3),(61.5 ± 15.4),(3 354.0 ±809.0),(6 888.4 ± 1 100.2),and (28.9 ± 4.6) pg/mL respectively;the peak values of IL-12p70 were (389.3 ± 118.1),(127.6 ± 10.0),(42.2 ± 3.5),(62.8 ± 8.4),and (4.8 ± 0.3) pg/mL respectively.Concentrations of MIP-1β and MIP-2 in E.coli and K.pneumoniae groups were significantly higher than other trial groups and control group (all P＜0.01),while concentrations of IL-12p70 in S.aureus and E.faecalis groups were both significantly higher than E.coli,K.pneumoniae,and control groups (all P＜0.01).Conclusion Concentrations of MIP-1β and MIP-2 in E.coli and K.pneumoniae groups were both significantly higher than those in S.aureus and E.faecalis groups,while concentrations of IL-12p70 in S.aureus and E.faecalis groups were both significantly higher than those in E.coli and K.pneumoniae groups.The combination detection of multiple cytokines or chemokines are valuable in predicting gram-positive or gram-negative bacterial infection,and can provide basis for treatment of early infection.

BACKGROUNDWith economic growth and urbanization there have been significant changes in the life style and diet of urban residents in large cities of China, which is experiencing a rapid increase in the prevalence of diabetes. While high prevalence of diabetes has been reported, little is known of the long-term effects of diabetes in such a large population. The aim of this study was to estimate the morbidity rate of diabetic peripheral neuropathy (DPN) in a Chinese urban diabetic population with more than 10 years' disease duration, and evaluate the relevant risk factors. The clinical manifestation of DPN and pain status was also assessed.

METHODSFive hundred and sixty-five diabetes patients were recruited into the study. Symptoms and examination helped diagnose neuropathy. The clinical manifestation of DPN was assessed with a visual analog pain score (VAS). Diabetic complication status was determined from medical records. Serum lipids and lipoproteins, glycosylated hemoglobin (HbA1c), and the urinary albumin excretion rate were measured.

RESULTSThe morbidity rate of DPN was 46.6%. HbA1c, hyperlipidemia, and retinopathy were significantly associated with neuropathy, and these risk factors were correlated with other diabetic micro and/or macrovascular complications. The average VAS pain score of the DPN patients was 4.12 ± 2.07. Severe and moderate pain was experienced by 11.4% and 40.5% respectively of DPN patients. About 3.7% of diabetic subjects had lower limb ulcer or amputation.

CONCLUSIONSThe morbidity rate of DPN for diabetic patients with > 10 years duration is very high compared to the range reported for other populations in the world. The risk factors for DPN include HbA1c, hyperlipidemia, and retinopathy. In long-standing diabetic patients, DPN was not associated with diabetic duration, and half of the DPN patients experienced considerable daily suffering.

Aged ; China ; Diabetic Neuropathies ; epidemiology ; metabolism ; physiopathology ; Female ; Glycated Hemoglobin A ; metabolism ; Humans ; Hyperlipidemias ; epidemiology ; metabolism ; physiopathology ; Male ; Middle Aged ; Pain ; etiology ; Risk Factors ; Urban Population

Objective:To evaluate the value of ultrasonographic features and classification defined by the Chinese-Thyroid Imaging Reporting and Data System (C-TIRADS) in differentiating follicular thyroid carcinoma (FTC) from follicular thyroid adenoma (FTA) .Methods:Data of 87 patients who underwent thyroid nodule-related surgery at Tianjin Medical University General Hospital from Jan. 2019 to Feb. 2023 and whose postoperative pathology confirmed FTC or FTA was divided into FTC group (26 cases) and FTA group (61 cases) were retrospectively analyzed. The differences of general data, pathological diagnosis, surgical methods, ultrasonographic features of thyroid nodules and C-TIRADS classification between the two groups were compared. Multivariate Logistic regression analysis was used to screen the independent predictors related to the occurrence of FTC, a regression model was established based on the independent predictors and a nomogram was drawn. Bootstrap method was used to verify the nomogram internally, and ROC curve and calibration curve were drawn to evaluate the diagnostic efficiency and predictive ability of the nomogram.Results:There were statistically significant differences between orientation, echo, composition, calcification, margin, halo and C-TIRADS classification between groups (all P<0.05) ; maximum diameter, echotexture, blood flow, general data, pathological diagnosis and surgical methods had no statistically significant differences between groups (all P>0.05) .Single-factor logistic regression analysis showed taller-than-wide, peripheral calcification, blurred/irregular edges or external invasion, C-TIRADS 4B/4C class were candidate variables for predicting the occurrence of FTC ( OR=10.909, 19.059, 19.259, 49.333, all P<0.05). Multivariate logistic regression analysis showed that peripheral calcification, blurry/irregular edges or extra-thyroid invasion, C-TIRADS 4B/4C class were independent predictors for the occurrence of FTC ( OR=33.731, 11.620, 22.794, all P<0.05) .The nomogram of FTC occurrence was constructed based on the above three independent predictors, and the AUC was 0.920 (95% CI=0.854-0.986), which showed that the nomogram had high diagnostic efficacy, and the optimal diagnostic threshold determined by the Youden index was 0.23 (corresponding to the total score of the nomogram is about 52 points), the sensitivity was 88.5%, and the specificity was 82.0%. The calibration curve drawn by internal verification was close to the reference line, suggesting that the nomogram had a high predictive ability. Conclusions:The nomogram prediction model for FTC occurrence based on the ultrasonographic features and classification defined by C-TIRADS has high diagnostic efficiency and predictive ability, and can help in the preoperative identification of FTC and FTA.

OBJECTIVETo explore the significance of apoptosis of rabbit small intestinal mucosal epithelial cells and lymphocytes, and lymphocytes of lumbrical process at early postburn stage.

METHODSTwenty-five Japanese white rabbits were randomly divided into 5 groups with 5 in each group, i.e. normal control (N), 3-postburn-hour group (3 PBH), 6 PBH, 12 PBH and 24 PBH groups. The rabbits in all PBH groups were inflicted with 30% TBSA III degree of flame burn on the back. The intestinal tissue samples were harvested from 5 anatomical sites for HE staining, electron microscopic examination and the detection of apoptosis in situ by TUNEL method at all the postburn time points. The results of TUNEL slides were analyzed statistically.

RESULTSHE staining revealed that there were relatively abundant apoptotic cells scattering solitarily in the lymph nodules and diffuse lymphatic tissue in the mucosal epithelial and mucosal lamina propria (and partially extended into the submucosal layer) of the intestine and lumbrical process in all burn groups. There were some disruption of intestinal mucosa in 24 PBH group. But no obvious inflammatory reaction and signs of necrosis were observed in all the slides. Apoptotic body formation could be identified by EM. Large number of blue-black positive cellular nuclei were revealed by TUNEL method with their distribution as similar to that found by HE staining. When comparing with those in control group, the apoptotic cells in small intestine and lumbrical process were increased obviously (P < 0.01) in 3 PBH group and reached the top level in 6 and 12 PBH groups (P < 0.01), declining thereafter to near value of 3 PBH in 24 PBH group, though it was still higher than control (P < 0.05). The number of apoptotic epithelial cells in middle distal portions of small intestinal mucosa in burn groups was much higher than that in proximal intestine (P < 0.05).

CONCLUSIONThere was a large number of apoptotic cells in rabbit small intestinal mucosal epithelium, gut associated lymphoid tissue and lymphocytes in the lumbrical process, especially in the middle and distal portions of the intestine. These change might be the cellular basis of postburn intestinal translocation of bacteria and endotoxin.

Animals ; Apoptosis ; Burns ; pathology ; Epithelial Cells ; pathology ; Female ; Intestinal Mucosa ; pathology ; Intestine, Small ; pathology ; Lymphocytes ; pathology ; Male ; Rabbits ; Time Factors

Objective:To study the influence of Mei mini maze procedure for atrial functional mitral regurgitation.Methods:The data of 33 patients with atrial fibrillation and atrial functional mitral regurgitation from January 2017 to June 2020 were retrospectively analyzed. All patients received Mei mini maze procedure for atrial fibrillation. The procedure is carried out thoracoscopically through the left thoracic approach. The ablation of atrial fibrillation includes bilateral circumferential pulmonary vein ablation, isolation of the left atrium posterior wall, left atrial appendage resection, ablation of Marshall's ligament and autonomic ganglion, etc. Follow-up was conducted by outpatient follow-up and telephone. Postoperative heart rhythm was recorded by the patient's symptoms, electrocardiogram, 24 h holter and other examinations. Postoperative mitral valve lesions were obtained by echocardiography.Results:33 patients successfully completed the operation. There was no conversion to thoracotomy and no perioperative death. Thirty patients(90.9%) maintained sinus rhythm at discharge. Before discharge, 16 patients had no mitral regurgitation in echocardiography, 8 patients had mild mitral regurgitation, and 9 patients had moderate mitral regurgitation. Follow-up was 1-4 years after discharge, with a mean of(2.6±1.1) years. Sinus rhythm was maintained in 23 patients(69.7%). 17 patients had no mitral regurgitation, 9 had mild mitral regurgitation, 6 had moderate, and 1 had severe mitral regurgitation. The degree of regurgitation in 25 patients was reduced compared with pre-operation, 5 patients remained unchanged, and 3 patients mitral regurgitation aggravated. Unreduced atrial functional mitral regurgitation was associated with recurrence of atrial fibrillation by Cox multivariate analysis.Conclusion:This study found a close relationship between atrial fibrillation rhythm and atrial functional mitral regurgitation. Most moderate atrial functional mitral regurgitation can be alleviated by effective treatment for atrial fibrillation. It is not recommended that patients with severe atrial functional mitral regurgitation only receive treatment for atrial fibrillation.

Objective To investigate the impact of the CYP3A4*1G and CYP3 A5*3 polymorphisms on the concentration/dose ratios of dilti-azem in kidney transplant patients.Methods Forty -seven cases of kidney transplant recipients were recruited.CYP3A4 and CYP3A5 geno-types were determined by polymerase chain reaction -restriction fragment length polymorphism.Plasma trough concentration of diltiazem was meas-ured by HPLC.The effects of CYP3A4*1G and CYP3A5*3 gene poly-morphisms on the concentration/dose ratios of diltiazem were evaluated and compared.Results There were fourteen cases of CYP3A4*1*1 (29.79%) homozygous genotype , twenty six cases of CYP3A4*1*1G (55.32%) heterozygous genotype , and seven cases of CYP3A4*1G*1G (14.89%) homozygous genotype.Allele frequencies of CYP3A4*1 and *1G were 57.45%and 42.55%, respectively.There were six ca-ses with CYP3A5*1*1 (12.77%), twenty one cases with CYP3A5*1*3(44.68%), and twenty cases with CYP3A5*3*3(42.55%).Allele frequencies of CYP3A5*1 and *3 were 35.11%and 64.89%, respectively.CYP3A4*1G gene polymorphisms were significantly related to the concentration /dose of diltiazem in kidney transplant patients.The diltiazem blood trough concentration/dose in CYP3A4*1*1 recipients was 1.52 times higher than that in patients with CYP 3A4*1G allelic genes.There was significant difference in diltiazem blood trough concen-tration/dose ratios between recipients with CYP3A4*1*1 genotypes and CYP3A4*1G carriers(P<0.05).CYP3A5*3 gene polymorphisms had slight impacts on diltiazem blood trough concentration /dose ratios but no difference was found among the CYP3A5 genotypes.Conclusion The CYP3A4*1G gene polymorphism is closely related to the blood con-centration/dose ratios of diltiazem in kidney transplant patients.

Objective:To compare the radiation chemistry effects on water molecules after ultra-high dose rate (FLASH) and conventional irradiation.Methods:Both FLASH and conventional irradiation were applied to ultrapure water, with the hydroxyl radical yield in the homogeneous phase detected using electron paramagnetic resonance (EPR) and the hydrogen peroxide (H 2O 2) yield in the diffusion phase analyzed uuxing fluorescence probe. The liposome model was then established to investigate the radiation chemistry effect of FLASH and conventional irradiation in inducing lipid peroxidation. Results:Radiation chemistry reactions were observed in water molecules after irradiation. In the homogeneous phase, the yield of free radicals using FLASH irradiation is similar to those from conventional irradiation ( P>0.05). In the diffusion phase, the amount of H 2O 2 produced by FLASH irradiation was significantly lower than those from conventional irradiation ( t=0.49-12.81, P<0.05). The liposome model confirmed that conventional irradiation could significantly induce lipid peroxidation through the radiation chemistry effect in water molecules as compared with FLASH irradiation ( t=0.31-11.73, P<0.05). Conclusions:The radiation chemistry effect in water molecules after FLASH irradiation was significantly lower than that from conventional irradiation. This could be one of the mechanisms of FLASH effect.

Objective To evaluate the safety of the split influenza vaccine Anflu.Methods A total of 198 subjects were enrolled in this open clinical trial.Of which,97 aged 18-60 years old and 101 aged 61 years old.28 of the subjects had history of chronic disease,while all the subjects were under stable health condition.After being infoma consented,each subject was vaccinated with one dose of split influeza vaccine and observed for 7 days for adverse reactions.Results A total of 17 subjects reported adverse reactions and the adverse reactions occurrence rate was 8.6%.The adverse reactions occurrence rate was 13.4%in the adult group and 4.0%in the elderly group(x2=5.620,P=0.018).Local adverse reactions occurrence rate was 5.5%and pain at the iniection site was the major symptom.System adverse reactions rate was 3.5% and fever was the most frequently reported symptom.None of the subjects with chronic disease history reported adverse reactions.Conclusion The results showed that the vaccine did not cause new adverse reaction,serious adverse event or rare adverse reaction.The split influenza vaccine was safe.

Objective To evaluate the safety of the split influenza vaccine Anflu.Methods A total of 198 subjects were enrolled in this open clinical trial.Of which,97 aged 18-60 years old and 101 aged 61 years old.28 of the subjects had history of chronic disease,while all the subjects were under stable health condition.After being infoma consented,each subject was vaccinated with one dose of split influeza vaccine and observed for 7 days for adverse reactions.Results A total of 17 subjects reported adverse reactions and the adverse reactions occurrence rate was 8.6%.The adverse reactions occurrence rate was 13.4%in the adult group and 4.0%in the elderly group(x2=5.620,P=0.018).Local adverse reactions occurrence rate was 5.5%and pain at the iniection site was the major symptom.System adverse reactions rate was 3.5% and fever was the most frequently reported symptom.None of the subjects with chronic disease history reported adverse reactions.Conclusion The results showed that the vaccine did not cause new adverse reaction,serious adverse event or rare adverse reaction.The split influenza vaccine was safe.