Objective To investigate the changes in cells on cell cycle,cell apoptosis and susceptibility to chemotherapeutic drugs of lung adenocarcinoma A549 cells under two dimensional (2D system) and three dimensional (3D system) culture conditions. Methods The three dimensional culture of A549 cells was carried out using rotatable culture bottle and desk-top water bath shake. Flow cytometer,in situ cell apoptosis detection kit,MTT and blood cell counter were employed to compare cell cycle,cell apoptosis and susceptibility to ADM of A549 cells under different culture conditions. Results The three dimensional culture system was confirmed to have been successful by the aid of observation under inverted and electron microscope. There were significant retardation of G_ 1 phase,lower cell apoptosis rate and decreased susceptibility to ADM in A549 cells in 3D system compared with those in 2D system. Conclusions There were remarkable differences in biological characteristics of A549 cells in two culture systems,indicating that A549 cells cultivated in 3D system simulated better solid tumor in vivo . The 3D system was very useful for further investigation of the behavior of solid tumor,so that anti-carcinoma chemotherapeutic drugs could be advantageously tested in vitro before clinical application.

Objective To explore the clinical value of combined detection of human epididymis protein 4(HE4) ,carbohydrate an‐tigen 199(CA199) and carbohydrate antigen 125(CA125) in diagnosis of ovarian cancer and benign ovarian cysts .Methods The se‐rum levels of HE4 ,CA125 and CA199 were determined by using chemiluminescent immunoassay in 45 cases of patients with ovarian cancer(ovarian cancer group) ,60 cases of patients with benign ovarian cyst(benign disease group) ,and 30 cases of healthy individu‐al(control group) .Results The serum levels of HE4 ,CA199 and CA125 in the ovarian cancer group were higher than those in the ovarian cancer group and control group ,there were statistically significant differences(P<0 .05) .Compared with the control group , the serum level of CA125 in benign disease group was significantly higher(P<0 .05) ,while no statistically significant differences of serum levels of HE4 and CA199 were observed between the two groups(P>0 .05) .The sensitivity of combined detection of the three indicators increased to 82 .2% ,and the specificity remained at 90 .0% .Conclusion Combined detection of HE4 ,CA125 and CA199 might have clinical significance in early diagnosis and identification of ovarian cancer .

Objective To study the effect of modified periocular injection on the pains induced by local application of drugs. Methods Sixty patients treated with periocular injection were divided randomly into experimental group and control group in equal number. The control group was treated with orthodox periocular injection,while experimental group with modified periocular injection, immersing the pinhead into lidocaine solution before injection.The pain level of patients was evaluated by self-designed pain scale. Result The pain level of modified periocular injection was lower than that of orthodox periocular injection statistically (Z=3.799, P<0.001).Conclusion Modified periocular injection can alleviate pains induced of local application of drugs obviously and increase their compliance,so it deserves clinically popularizing.

Objective:To summarize the oblique branches found in the transfer of anterolateral thigh flap (ALTF) and to handle the issues of oblique branch.Methods:Thirty patients who require surgery of ALTF transfer from May, 2017 to October, 2019 were enrolled. CTA examination was perfected prior surgery and the origin of ALTF vessels was preliminarily determined. During the surgery, Three-longitudinal-and-five-transverse methods were used to locate and design the flap. The ALTF was taken according to the flap design. Attention should be paid to the location of the oblique branch and the separation of the vessels of oblique branch to avoid a damage as much as possible. The vessels of oblique branch should be completely explored and separated. One or 2 vascular pedicles were cut according to whether the oblique branch vessels and the descending branch vessels were joined together during surgery. Clamping tests were carried out on the 2 vascular pedicles to determine an arterial blood supply. Super drainage of vascular pedicle veins according to the situation of blood circulation. After the surgery, routine treatment was carried out. Blood supply, skin temperature, swelling degree, exudation and survival of the flap were closely observed and regular follow-up was carried out.Results:Among the 30 ALTF examined by CTA, 13 patients were identified with oblique branch vessels before operation. During operation, 11 oblique branch vessels (The occurrencce rate was 36.6%)were found to enter the flap, and were completely preserved. Of the 11 identified oblique branch vessels, 8 had 2 vascular pedicles taken and the vascular pedicles were treated by venous super drainage technique. The postoperative blood supply of the flap was good; The skin temperature was closed to surrounding normal skin; Swelling of flap was minor and there was little seepage. The flaps all survived after surgery with stage one healing. Followed-up time was 3-32 (average 16.1) months. The recipient site healing was good, and the function and appearance were satisfactory. The joint movement at the donor site was normal, and there was no obvious loss of local sensation.Conclusion:More than one third of the oblique branches appear in this group. The oblique branch vessels should be preserved as much as possible to avoid issues in relation to the oblique branch. Reasonable handling of oblique branch is the key to the success of the surgery.

Objective To analyze the epidemiological characteristics of malaria in Hechi City,Guangxi Zhuang Autono?mous Region from 2005 to 2015,so as to provide the evidence for formulating and adjusting measures of malaria elimination. Methods The data of malaria cases in 11 counties of Hechi City from 2005 to 2015 were collected and analyzed by using Micro?soft Office Excel 2003 and SPSS 17.0. Results A total of 160 malaria cases were reported in Hechi City from 2005 to 2015, and the annual average malaria incidence was 3.6 per million. There were 10 local recurrence cases(accounting for 6.25%,10/160)and 150 imported cases(accounting for 93.75%,150/160). For the etiology,Plasmodium vivax accounted for 51.87%(83/160)in these cases,P. falciparum accounted for 34.38%(55/160),P. ovale accounted for 1.25%(2/160),P. malariae account?ed for 5.00%(8/160),and the indeterminate accounted for 7.50%(12/160). During the period of 11 years,the malaria inci?dence first dropped and then rose. There were no local cases after 2009. However,an imported falciparum malaria death case was reported in 2010,a severe imported falciparum malaria case was reported in 2013 and another in 2015. A severe case of ma?ternal?neonatal vivax malaria was reported in 2014. These cases were mainly distributed in 10 counties of Hechi City,with more young male adults who engaged in digging mining. More cases concentrated from April to August. Most of the reported malaria cases were imported,60.00%(96/160)of them returned from Africa and Southeast Asia,and 33.75%(54/160)from other do?mestic provinces. The median of the interval from symptom appearance to diagnosis was 5 d,and there was a significant differ?ence among the above years(c2=33.40,P<0.05). Conclusions Malaria is still an important public health problem in Hechi City,and the appropriate control measures and effective tools should be strengthened for malaria elimination. The key to consoli?date the achievements of malaria control is strengthening the malaria monitoring management of the floating population.

Objective Background and objective The Cox maze Ⅲ procedure has been considered the gold standard for the surgical treatment of atrial fibrillation (AF) and the modified Cox maze use bipolar radiofrequency ablation instead of the cut-and-sew technique,while also reducing the connecting lines in left atrium.This study was to understand whether completely isolation the posterior left atrium by increasing left atrium ablation lines can enhance the survival after modified Cox maze procedure.Methods From Jan 2009 to Dec 2009,all the patients underwent the Cox maze procedure,following the same examination,were divided into two groups (case-control): box lesion group (n =60),which is means the right and lefi pulmonary vein lesions were connected inferiorly by adding ablation lines,thereby completely isolating the posterior left atrium and the non-box lesion group (n =60).Similar interventions were given during the perioperative periods.Comparing the risk factors before surgeries,then follow-up was 100％ complete,and the mean follow-up was 9 ± 8.4 months.Results Except the box lesion had long time of extracorporeal circulation,the characteristics of the 2 groups were similar because there were no differences in age,AF duration,left atrial diameter,left ventricular ejection fraction or homochronous operation between the groups and there was no operative death.No patient was lost to follow-up.In the mean follow-up duration,a patient was died of intracerebral hemorrhage at 2 months postoperationly in the box lesion group and a patient suffered from stroke at 12 months postoperationly in the non-box lesion.The overall freedom from AF recurrence was higher in the box lesion group at 1 (70％ vs 51.2％,P =0.039) and 3 (78.3％ vs 60％,P =0.030) months.While it was 80％ vs 71.7％ (P =0.286) at 6 months.Eight patients wrere readmitted because of the recurrence of AF,4 and 2 patients underwent catheter ablation in box and non-box group respectively,one patient in each group underwent electrical conversion.After treatment,all of the patients were recovered to sinus rhythm and continued to take cordarone.Conclusion Compared to the single connecting between right and left pulmonary vein,isolating the entire posterior left atrium by creating a box lesion showed higher freedom from AF in the earlier months (1-3months).However,half a year after the surgery,there were no significant differences between the two groups.

Background Lattice corneal dystrophy (LCD) is a progressive disease,whose clinical features are varied in different stages.It is rarely be reported that clinical findings of different stages and factors of promoting the occurrence and development on LCD in a family.Objective The aim of this study was to identify the characteristics of the pedigree and clinical features of different stages in a LCD family,and further to discuss its influence factors.Methods A cross-sectional study was performed in this study.A Chinese family with LCD was enrolled in Shenzhen Eye Hospital from 2015 to 2016.Questionnaires for disease-related history,visual acuity measurement,ocular anterior segment examination and color photography were carried out for all the members of the family.In addition,anterior segment OCT (AS-OCT),laser scanning confocal microscope and corneal endothelium microscope were used to observe the morphology of corneal stroma and changes of corneal endothelial cells.The pedigree chart was drawn by Cyrillic2.1 software and analyzed based on Mendel law.Results This family included 5 generations of 73 members.Patients with LCD were found in each generation with similar morbidity in different gender,which followed the law of autosomal dominant inheritance.Eleven patients were found in 49 members related with Ⅲ1 of this family with the prevalence rate of 22.45％ and onset age at 21-50 years old,and the course of disease was 3-34 years.All of the members had no systemic disease except for two patients (Ⅲ 1 and Ⅲ 5) with hypertension.In the early stage of LCD,some bifurcate striolae appeared in the patients' corneal stroma without symptoms for many years.In the progressive stage,there was corneal irritation symptom accompanying with vision's decrease in the eyes with LCD.The bifurcate striolae were increased,widened and interwoven into lattice lines that the boundaries gradually became fuzzy,then corneal macula was formed because of recurrent corneal infiltration,and eventually resulted in corneal leucoma.High reflection corresponding to the pathologic region was showed by laser scanning confocal microscope and AS-OCT.No significant differences were found in corneal endothelial cell density and the percentage of hexagonal cells between LCD patients and normal phenotype families (t =1.887,P=0.075;t=-0.719,P =0.481).Penetrating keratoplasty was performed in a patient with corneal opacity and serious corneal opacity occurred near the surgical incision one year after the surgery.One patient was diagnosed as LCD 2 years after laser assisted in-situ keratomileusis.One patient was a welder.Conclusions LCD is autosomal dominant inheritance in the family.The clinical manifestations of LCD in the early,progressive and late stage can be seen in the pedigree,which offers a reference for ophthahnologists.Corneal surgery and lesion may induce the onset or aggravation of LCD.

Although the surgical techniques has already improved in heart transplantation, heart preservation is still the biggest obstacle to the surgery.At present, heart preservation effective time is only 4-6 hour.How to extent the time of heart preservation is a major research direction.Comparison of available preservations for heart transplantation based on its mechanism and the prospect of its Clinical application.

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Objective Heart transplantation is an effective treatment of end-stage heart diseases and extending the time of donor heart preservation helps to make up for the shortage of donor hearts. This study was to investigate whether high-pressured mixed gas ( HPMG) of carbon monoxide and oxygen could prolong the time of donor heart preservation and its mechanisms. Methods Forty-eight C57BL/6 male mice aged 4-6 weeks were randomly divided in-to four groups of equal number:control ( the donor heart isolated but not transplanted) , immediate transplantation ( the donor heart transplanted right after isolated) , HTK-preservation ( the donor heart preserved in histidine-tryptophan-ketoglutarate solution for 24 hours after isolated, and HPMG preservation ( the donor heart preserved in an HPMG chamber with the oxygen partial pressure of 3200 hPa and carbon monoxide partial pressure of 800 hPa for 24 hours after isolated) .Another 36 recipient mice aged 6-8 weeks were randomly assigned to receive the donor heart immediately after harvested (n=12), preserved in HTK solution (n=12), or preserved in HPMG (n=12).At 2 hours after transplantation, the status of heart re-beating and cardiac function were compared among different groups of recipient mice.At 24 hours, tissues were taken from the transplanted hearts for examination of pathologic changes by HE stai-ning, detection of the apoptosis of cardiac cells by TUNEL, and determination of the expressions of microtubule-associated protein 1 light chain 3 -Ⅱ(LC3-Ⅱ) and B cell lymphoma/leukemia-2 (Bcl-2) by Western blot. Resul ts The re-beating rates of the imme-diately transplanted and HPMG-preserved hearts were significantly higher than that of the HTK-preserved ones (P<0.05).At 2 hours after transplantation, the cardiac function scores were 2.5 (2.0-2.9), 0.8 (0.5-1.0), and 4.5 (4.0-4.5) in the immediate implantation, HPMG-preservation and HTK-preservation groups respectively, with statistically significant differences between any two groups (P<0.05).The expressions of LC3-Ⅱand Bcl-2 were 2.06 ±0.29 and 0.87 ±0.18 in the HPMG-preserved heart recipients and 1.24 ±0.20 and 2.07 ±0.32 in the immediately transplanted heart recipients, both higher than 0.13 ±0.03 and 0.19 ±0.02 in the controls and 0.16 ±0.06 and 0.26 ±0.08 in the HTK-preserved heart recipients (P<0.05), the Bcl-2 higher in the HTK-pre-served heart recipients than in the controls (P<0.05), and the LC3-Ⅱ expression higher in the HPMG-preserved heart recipients than in the immediately transplanted heart recipients (P<0.05).HE staining showed that cell edema and inflammatory cell infiltration were more obvious in the HPMG-preserved heart recipients than in the controls and immediately transplanted heart recipients but less obvious than in the HTK-preserved heart recipients.The rate of cell apoptosis was dramatically increased in the HPMG-and HTK-pre-served heart recipients ([5.04 ±1.77]%and [26.72 ±5.23]%) in comparison with the controls ([1.08 ±0.56]%) (P<0.01) and immediately transplanted heart recipients ([2.13 ±1.71]%) (P<0.01) but decreased in the HPMG as compared with the HTK-preserved heart recipients (P<0.01). Conclusion High-pressured mixed gas preservation can reduce cold ischemia-reperfu-sion injury of the donor heart, which may be associated with its promotion of autophagy, provision of energy to cells, and apoptosis of cardiocytes in the donor heart.