Objective:To investigate the efficacy of endoscopic histoacryl injection in cirrhotic patients with newly-developed esophagogastric varices (EGV) who have previously undergone splenectomy combined with pericardial devascularization.Methods:From January 2015 to January 2020, 125 cirrhotic patients with EGV treated with endoscopic histoacryl injection at the Department of Gastroenterology, Jinling Hospital, Medical School of Nanjing University, were included in the retrospective analysis. There were 45 patients in the group of splenectomy combined with pericardial devascularization (splenectomy group for short) and 80 patients in the non-splenectomy group. The efficacy of endoscopic treatment, postoperative variceal improvement, rebleeding rate, and complications were analyzed between the two groups.Results:Endoscopic histoacryl injection was successfully completed in all 125 patients, and the median volume of histoacryl was 4.5 mL. The overall effective rate in splenectomy and non-splenectomy group was 80.0% (36/45) and 57.5% (46/80), respectively. The difference in the number of significantly effective, effective, and ineffective cases between the two groups was statistically significant (16, 20, 9 cases, and 20, 26, 34 cases, respectively, χ 2=6.469, P=0.039). Two and 14 patients developed rebleeding in the splenectomy group and non-splenectomy group, respectively; and the difference in the rebleeding rate between the two groups was statistically significant (4.4% VS 17.5%, Log-rank P=0.039). No patient died within 1 year in either group, and no serious complications such as ectopic embolism occurred. Conclusion:After splenectomy combined with pericardial devascularization in cirrhotic patients with EGV and hypersplenism, the application of histoacryl has better short-term efficacy and can significantly reduce the rebleeding rate compared with the non-splenectomy group.

Objective:To investigate the clinical efficacy and prognostic influencing factors of radical surgery for duodenal gastrointestinal stromal tumor (GIST).Methods:The retrospective cohort study was conducted. The clinicopathological data of 741 duodenal GIST patients who under-went radical surgery in 17 medical centers, including 121 cases in Renji Hospital Affiliated to Shanghai Jiaotong University School of Medicine, 121 cases in Chinese PLA General Hospital, 116 cases in Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 77 cases in Fudan University Shanghai Cancer Center, 77 cases in West China Hospital, Sichuan University, 31 cases in Guangdong Provincial People′s Hospital, 24 cases in Fujian Cancer Hospital, 22 cases in Fujian Medical University Union Hospital, 25 cases in Shengjing Hospital of China Medical University, 19 cases in Xiangya Hospital, Central South University, 23 cases in Zhejiang Cancer Hospital, 17 cases in Liaoning Cancer Hospital＆Institute, 17 cases in the First Affiliated Hospital of Xiamen University, 15 cases in Sun Yat-sen University Cancer Center, 14 cases in the First Affiliated Hospital of Nanjing Medical University, 14 cases in Zhongshan Hospital Affiliated to Xiamen University and 8 cases in General Hospital of Chinese People′s Liberation Army Air Force, from January 2010 to April 2020 were collected. There were 346 males and 395 females, aged 55(range, 17?86)years. Observation indicators: (1) neoadjuvant treatment; (2) surgical and postoperative situations; (3) follow-up; (4) stratified analysis. Follow-up was conducted using outpatient examination or telephone interview. Patients were followed up once every 3?6 months during neoadjuvant therapy and once every 6?12 months after radical surgery to detect tumor recurrence and survival of patient up to April 2022. Measurement data with normal distribution were represented as Mean± SD. Measurement data with skewed distribution were represented as M(range), and comparison between groups was conducted using the Mann-Whitney U test. Count data were described as absolute numbers or percentages, and comparison between groups was conducted using chi-square test or Fisher exact probability. The Kaplan-Meier method was used to draw survival curves and calculate survival rates. Log-rank test was used for survival analysis. The COX regression model was used for univariate and multivariate analyses. Propensity score matching was done by the 1∶1 nearest neighbor matching method, and the matching tolerance was 0.02. Results:(1) Neoadjuvant therapy. Of the 741 patients, 34 cases received neoadjuvant therapy for 8(range, 3?44)months. Cases assessed as partial response, stable disease and progressive disease before the radical surgery of the 34 cases were 21, 9, 4, respectively. The tumor diameter of the 34 patients before the neoadjuvant therapy and before the radical surgery were 8.0(range, 3.0?26.0)cm and 5.3(range, 3.0?18.0)cm, with the regression rate as 31.9%(range, ?166.7% to 58.3%). (2) Surgical and postoperative situations. Of the 741 patients, 34 cases underwent radical surgery after receiving neoadjuvant therapy, and 707 cases underwent radical surgery directly. All the 741 patients underwent radical surgery successfully, in which 633, 102 and 6 cases received open surgery, laparoscopic surgery and endoscopic treatment, respectively. Of the 633 cases receiving open surgery and the 102 cases receiving laparoscopic surgery, cases with surgical resection range as pancreatoduodenectomy (PD) was 238, and cases with surgical resection range as duodenal limited resection, including duodenal wedge resection, distal gastrectomy, segmental duodenal resection, local resection of duodenal tumor or segmental duodenum combined with subtotal gastrectomy, was 497, 226, 55, 204, 12. Of the 741 patients, 131 cases had post-operative complications including 113 cases with grade Ⅰ?Ⅱ complications and 18 cases with ≥ grade Ⅲ complications of the Clavien-Dindo classification. The duration of postoperative hospital stay of the 741 patients was 13(range, 4?120)days. Of the 707 patients receiving direct radical surgery, 371 cases were evaluated as extremely low risk, low risk, medium risk of the modified National Institutes of Health (NIH) risk classification after surgery, and 336 cases were evaluated as high risk in which 205 cases receive postoperative adjuvant imatinib therapy with the treatment time as 24(range, 6?110)months. (3) Follow-up. All the 741 patients were followed up for 58(range, 7?150)months. During the follow-up, 110 patients had tumor recurrence and metastasis. The 1-, 3-, 5-year overall survival rates and 1-, 3-, 5-year disease-free survival rates of the 741 patients were 100.0%, 98.6%, 94.5% and 98.4%, 90.9%, 84.9%, respectively. The 1-, 3-, 5-year overall survival rates and 1-, 3-, 5-year disease-free survival rates of the 707 patients receiving direct radical surgery were 100.0%, 98.5%, 94.3% and 98.4%, 91.1%, 85.4%, respectively. (4) Stratified analysis. ① Analysis of prognostic factors in patients undergoing radical surgery directly. Results of univariate analysis showed that primary tumor location, tumor diameter, mitotic count, modified NIH risk classification and tumor gene information were related factors affecting the overall survival of 707 patients with primary duodenal GIST who underwent direct radical surgery ( hazard ratio=0.43, 0.18, 0.22, 0.06, 0.29, 95% confidence intervals as 0.20?0.93, 0.09?0.35, 0.10?0.50, 0.03?0.12, 0.09?0.95, P<0.05). The primary tumor location, tumor diameter, mitotic count, modified NIH risk classification were related factors affecting the disease-free survival of 707 patients with primary duodenal GIST who underwent direct radical surgery ( hazard ratio=0.65, 0.25, 0.25, 0.10, 95% confidence intervals as 0.41?1.03, 0.17?0.37, 0.15?0.42, 0.07?0.15, P<0.05). Results of multivariate analysis showed that primary tumor located at the horizontal segment of duodenum, mitotic count >5/50 high power field, tumor gene KIT exon 9 mutation were independent risk factors affecting the overall survival of 365 patients with primary duodenal GIST after removing 342 patients without tumor gene information who underwent direct radical surgery ( hazard ratio=2.85, 2.73, 3.13, 95% confidence intervals as 1.12?7.20, 1.07?6.94, 1.23?7.93, P<0.05). Tumor diameter >5 cm and mitotic count >5/50 high power field were independent risk factors affecting the disease-free survival of 707 patients with primary duodenal GIST who underwent direct radical surgery ( hazard ratio=3.19, 2.98, 95% confidence intervals as 2.05?4.97, 1.99?4.45, P<0.05). ② Effect of postoperative adjuvant therapy on prognosis of high-risk patients of modified NIH risk classification. Of the 336 patients evaluated as high risk of the modified NIH risk classification, the 5-year overall survival rate and 5-year disease-free survival rate were 94.6% and 77.3% in the 205 cases with postoperative adjuvant therapy, versus 83.2% and 64.4% in the 131 cases without postoperative adjuvant therapy, showing significant differences between them ( χ2=8.39, 4.44, P<0.05). Of the 205 patients evaluated as high risk of the modified NIH risk classification who received postoperative adjuvant therapy, there were 106 cases receiving postoperative adjuvant therapy <36 months, with the 5-year overall survival rate and 5-year disease-free survival rate were 87.1% and 58.7%, and there were 99 cases receiving post-operative adjuvant therapy ≥36 months, with the 5-year overall survival rate and 5-year disease-free survival rate were 100.0% and 91.5%. There were significant differences in the 5-year overall survival rate and 5-year disease-free survival rate between the 106 patients and the 99 patients ( χ2=13.92, 29.61, P<0.05). ③ Comparison of clinical efficacy of patients with different surgical methods. Before propensity score matching, cases with primary tumor located at bulb, descending, horizontal, ascending segment of duodenum, cases with tumor diameter ≤5 cm and >5 cm were 95, 307, 147, 34, 331, 252, in the 583 patients receiving open surgery with complete clinical data, versus 15, 46, 17, 5, 67, 16 in the 83 patients receiving laparoscopic surgery with complete clinical data, showing no significant difference in the primary tumor location ( χ2=0.94, P>0.05), and a significant difference in the tumor diameter ( χ2=17.33, P<0.05) between them. After propensity score matching, the above indicator were 16, 39, 20, 8, 67, 16 in the 83 patients receiving open surgery, versus 15, 46, 17, 5, 67, 16 in the 83 patients receiving laparoscopic surgery, showing no significant difference between them ( χ2=1.54, 0.00, P>0.05). Cases with postoperative complications, cases with grade Ⅰ?Ⅱ complica-tions and ≥grade Ⅲ complications of the Clavien-Dindo classification, duration of postoperative hospital stay, the 5-year overall survival rate and 5-year disease-free survival rate were 17, 12, 5, 11(range, 5?120)days, 92.0%, 100.0% in the 83 patients receiving open surgery, versus 9, 7, 2, 11(range, 5?41)days, 91.6%, 97.3% in the 83 patients receiving laparoscopic surgery, showing no signi-ficant difference in postoperative complications, duration of postoperative hospital stay, the 5-year overall survival rate and 5-year disease-free survival rate ( χ2=2.91, Z=3 365.50, χ2=3.02, 1.49, P>0.05) between them. There was no significant difference in complications of the Clavien-Dindo classification between them ( P>0.05). ④ Comparison of clinical efficacy of patients with primary tumor located at the descending segment of duodenum who underwent surgery with different surgical resection scopes. Before propensity score matching, cases with tumor diameter ≤5 cm and >5 cm, cases with tumor located at opposite side of mesangium and mesangium were 71, 85, 28, 128 in the 156 patients with primary tumor located at the descending segment of duodenum who underwent PD with complete clinical data, versus 92, 41, 120, 13 in the 133 patients with primary tumor located at the descending segment of duodenum who underwent duodenal limited resection with complete clinical data, showing significant differences between them ( χ2=16.34, 150.10, P<0.05). After propensity score matching, the above indicator were 28, 13, 16, 25 in the 41 patients with primary tumor located at the descending segment of duodenum who underwent PD with complete clinical data, versus 28, 13, 16, 25 in the 41 patients with primary tumor located at the descending segment of duodenum who underwent duodenal limited resection with complete clinical data, showing no significant difference between them ( χ2=0.00, 0.00, P>0.05). Cases with postopera-tive complications, cases with grade Ⅰ?Ⅱ complications and ≥grade Ⅲ compli-cations of the Clavien-Dindo classification, duration of postoperative hospital stay, the 5-year overall survival rate and 5-year disease-free survival rate were 13, 11, 2, 15(range, 9?62)days, 94.2%, 64.3% in the 41 patients with primary tumor located at the descending segment of duodenum who underwent PD with complete clinical data, versus 9, 8, 0, 15(range, 7?40)days, 100.0%, 78.8% in the 41 patients with primary tumor located at the descending segment of duodenum who underwent duodenal limited resection with complete clinical data, showing no significant difference in post-operative complica-tions, the 5-year overall survival rate and 5-year disease-free survival rate ( χ2=0.99, 0.34, 1.86, P>0.05) between them. There was no significant difference in complications of the Clavien-Dindo classification ( P>0.05) and there was a significant difference in duration of postopera-tive hospital stay ( Z=614.50, P<0.05) between them. Conclusions:The clinical efficacy of radical surgery for duodenal GIST are ideal. Primary tumor located at the horizontal segment of duodenum, mitotic count >5/50 high power field, tumor gene KIT exon 9 mutation are independent risk factors affec-ting the overall survival of patients undergoing direct radical surgery and tumor diameter >5 cm and mitotic count >5/50 high power field are independent risk factors affecting the disease-free survival of patients. There is no significant difference in the short-term efficacy and long-term prognosis between patients undergoing open surgery and laparoscopic surgery. For patients with primary tumor located at the descending segment of duodenum, the duration of postoperative hospital stay is longer in patients undergoing PD compared with patients undergoing duodenal limited resection. For patients evaluated as high risk of the modified NIH risk classification, posto-perative adjuvant therapy and treatment time ≥36 months are conducive to improving the prognosis of patients.

Objective To investigate the predictive value of serum adiponectin (APN) for post-stroke depression (PSD) in patients with ischemic stroke. Methods Patients with acute ischemic stroke admitted to the Departments of Neurology, the Affiliated Brain Hospital of Guangzhou Medical University, and Dongguan Changping Hospital were enrolled prospectively from September 2016 to January 2018. Serum APN levels were measured by radioimmunoassay the next day after admission. PSD was diagnosed by using the Diagnostic and Statistical Manual of Mental Disorders (4th Edition) at one month after discharge. The baseline clinical features in the PSD group and the non-PSD group were compared. Multivariable logistic regression analysis was used to determine the risk factors for PSD. The receiver operating characteristic (ROC) curve was used to analyze the predictive value of APN for PSD. Results A total of 220 patients were enrolled, including 57 (25.9%) in the PSD group and 163 (74.1%) in the non-PSD group. The proportion of patients with diabetes (P=0.020) and living alone (P=0.012), as well as age (P=0.17), homocysteine (P=0.009), C-reactive protein ( P=0.001), and National Institutes of Health Stroke Scale score ( P=0.007) in the PSD group were significantly higher than those in the non-PSD group, while the APN level was significantly lower than that in the non-PSD group (P=0.003). Multivariate logistic regression analysis showed that the 4th quartile of serum APN level was used as a reference, the 1st quartile of APN level was an independent risk factor for PSD (odds ratio, 4 .202, 95% confidence interval 1.401-12.067; P=0.013). ROC curve analysis showed that the area under the curve of serum APN level for predicting PSD was 0.642 (95% confidence interval 0.564-0.721; P=0.001). The optimal cutoff value was 6.4 mg/L, and its sensitivity of predicting PSD was 63.2%, the specificity was 63.8%, the positive predictive value was 83.3%, and the negative predictive value was 57 .7%. Conclusion Serum APN has a certain predictive value for PSD in patients with ischemic stroke.

Objective To verify the targeting regulatory relationship between microRNA -939 (miR -939) and CD2 -associated protein (CD2AP).Methods The online RegRNA software was used to predict the human CD2AP promoter for potential binding sites complementary to miR -939.HEK -293T cells were cotransfected with hu-man CD2AP promoter plasmid pGL3 -2K and microRNA negative control (miR -NC)or miR -939 mimics,and the relative luciferase activity(RLA)was detected at 24 h post -transfection.HEK -293T cells were transfected with miR -NC or miR -939 mimics for 48 h,and the CD2AP mRNA expression level was detected by adopting reverse tran-script and real -time fluorescence quantification -PCR,while the CD2AP protein expression level was detected by using Western blot.Results (1 )There were 2 miR -939 binding sites at CD2AP promoter region,located at -468 to -491 and -654 to -677 upstream of initiation codon ATG (marked as +1 )relatively.(2)At 30 nmol/L,50 nmol/L,the RLA in miR -NC group and miR -939 group were 6.81 ±0.88 vs 6.07 ±2.24,5.88 ±1 .44 vs 3.94 ± 0.79 relatively,and there were no significant differences between the 2 groups (t =3.04,2.06,all P >0.05),while the RLA between the 2 groups were 5.58 ±0.58 vs 3.29 ±0.64 at 1 00 nmol/L,and the difference was significant between the 2 groups(t =4.07,P <0.05).(3)At 30 nmol/L,50 nmol/L and 100 nmol/L,the relative CD2AP mRNA expression in miR -NC group and miR -939 group were 1 .00 ±0.01 vs 0.80 ±0.08,1 .00 ±0.00 vs 0.80 ±0.1 3 and 1 .00 ± 0.00 vs 0.72 ±0.07 relatively,while the CD2AP mRNA expression was decreased by 20% -30% at each concentration level,and there were significant differences between the 2 groups (t =4.44,2.93,6.84,all P <0.05).(4)At 50 nmol/L, the relative CD2AP protein expression in miR -NC group and miR -939 group were 0.48 ±0.09 vs 0.19 ±0.12,and the CD2AP protein expression was decreased,and the difference was significant (t =3.36,P <0.05).Conclusions CD2AP is the target gene of miR -939,and miR -939 can down -regulate the expression of CD2AP both in mRNA and protein levels by targeting its promoter region,which indicates that miR -939 may mediate the podocyte injury.

Objective To explore an improving primary culture method of human pituitary adenoma cells.Methods Thirty-six pituitary adenoma specimens,collected from excision and conformed by pathology in our hospital from November 2014 to June 2015,were used.Based on the former experience from papers and ourselves,our primary culture methods of several different kinds of human pituitary adenomas were improved.Purification and passage culture of cells were performed in the following experiments.CCK-8 assay was used to detect the multiplication capacity of pituitary adenoma cells;immunofluorescent staining and Western blotting were used to observe the cytokeratin and vimentin protein expressions.Results By using the improve culture methods,tumor cells proliferated in suspension with good status and had the tendency of aggregation in culture medium.The proliferation test showed that the tumor cells enjoyed proliferation ability in vitro;tumor cells could pass over five generations.The growth curve of pituitary adenoma cells showed S pattern.Immunofluorescent staining and Western blotting indicated negative cytokeratin and vimentin protein expressions,showing that the cells were from epithelium.Conclusion Each type of pituitary adenoma cells has characteristics of suspended growth and tendency of gathering by using this improved method.

Objective To study the application value of the combined detection of serum HE4 ,CAl25 ,CA72‐4 and IL‐6 in the di‐agnosis of ovarian malignant tumor .Methods The serum levels of CA125 and CA72‐4 were determined by ECLI ,and the serum levels of HE4 and IL‐6 were determined by ELISA in 32 patients with ovarian cancer ,56 patients with benign ovarian disease and 40 healthy controls .The detection results were performed the statistical analysis .Results The serum levels of HE4 ,CAl25 ,CA72‐4 and IL‐6 in the ovarian cancer group were significantly higher than those in the other two groups with statistically significant differ‐ences(P< 0 .01) ;the sensitivity of CA125 detection in the single index detections was highest(75 .0% ) ,the specificity of HE4 was highest(83 .9% ) ,the highest diagnostic efficiency of single index detection was 79 .5% .The sensitivity ,specificity and efficiency of the combined detection for diagnosis were 96 .9% ,71 .4% and 80 .7% .Except the specificity ,the sensitivity and diagnostic efficien‐cy of the combined detection were higher than those of any one of single index detection .Conclusion The combined detection of se‐rum HE4 ,CAl25 ,CA72‐4 and IL‐6 could increase the diagnostic efficiency of ovarian cancer and conduces to the diagnosis and help to the diagnosis and differential diagnosis of ovarian malignancies .

Objective To develop a newly transparent sheath and explore the application value of the sheath in neuroendoscopic intra-axial surgery Methods Sixty-eight patients,admitted to our hospital from August 2010 to February 2013,underwent endoscopic surgery with the help of new sheath.Pre-operated CT or MRI were performed to locate the puncture direction;the transparent sheath was used to provide a trajectory in the brain parenchyma,and offer the clear vision surround the sheath tube and the puncture direction.Results Forty-eight patients adopted lateral ventricle anterior horn approach and 46 (95.83％) achieved lesions in the first puncture.Three patients with septum pellucidum cyst and one with Monro obstruction were successfully received the cystic wall fenestration or recanalization with the sheath directly.After the entry point were determined with CT in 10 intracerebral hematoma,the sheath was inserted into the hemorrhage cavity simply and easily;and a smash-suction tube was inserted to evacuate the hematoma safely.Conclusion The transparent sheath can be inserted into lateral ventricle,hemorrhage cavity or cyst efficiently,and provide perfect surgical vision around the sheath and the puncture direction to estimate the brain parenchyma,hemorrhage cavity or the hemorrhage remains;therefore,this transparent sheath,as a safely neuroendoscopic intra-axial surgical instrument,is deserved to be spread.

Objective-To-investigate-surgical-skills-and-safety-of-laparoscopic-hysterectomy-procedure-for-enlarged-uterus-in-woman-with-adenomyosis-.-Methods-Fourteen-cases-of-adenomyosis-with-uterus-larger-than-twelve-week-pregnancy-accepted-laparoscopic-hysterectomy-form-December-2012-to-December-2013-in-our-hospital-.-Some-operating-techniques-,-including-the-management-of-uterine-vessels-and-separation-of-cardinal-ligament-and-uterosacral-ligament-,-were-modified-.-Results-The-operation-was-performed-successfully-under-laparoscopy-in-all-the-cases-.No-conversion-to-open-surgery-was-required-.No-operating-complication-happened.The-operating-time-was-(160.7-±66.1)-min,-and-the-intraoperative-blood-loss-was-(184.3-±50.5)-ml.-Conclusion-Modified-laparoscopic-techniques-should-be-considered-as-an-acceptable-alternative-in-hysterectomy-for-the-management-of-large-uterus-in-woman-with-adenomyosis-.

Objective To explore the role of Th17-and Treg-derived catecholamines during collagen-induced ar?thritis (CIA) progression. Methods Eighteen male DBA/1 mice were randomly divided into control group, CIA model groupⅠ(day 35) and CIA model groupⅡ(day 55). The CIA models were induced by typeⅡcollagen (CⅡ) injection from tails. mRNA expression of Th17 specific transcription factor include ROR-γt, cytokines, IL-17 IL-22, Treg specific transcription factor, Foxp3, cytokines, TGF-βand tyrosine hydroxylase (TH) in lymph nodes were examined by real-time PCR. Co-local?ization of ROR-γt or Foxp3, with TH, vesicular monoamine transporter-2 (VMAT-2) or monoamine oxidase (MAO) in lymph nodes were observed by immunofluorescence staining. Results In lymph nodes of mice in CIAⅠgroup and CIAⅡgroup, mRNA expression of ROR-γt, IL-17, TH and IL-22 were upregulated, while mRNA expression of Foxp3 and TGF-βex?pression was downregulated compared to those expression in control group. The upregulated expression of IL-17 was signifi?cantly reduced in CIAⅡgroup compared with that in CIAⅠgroup. In the lymph nodes of both intact and CIA mice, co-lo?calization of ROR-γt or Foxp3 with TH, VMAT-2 or MAO was seen in some cells. The numbers of cells that are double-pos?itive of ROR-γt/TH,ROR-γt/VMAT-2 and ROR-γt/MAO IL-17 were increased in CIA groups compared to those in con?trol group. And they are significantly reduced in CIAⅡgroup compared with those in CIAⅠgroup. Conclusion The abili?ty to synthesize catecholamines in Th17 cells was increased in lymph node in mice from CIA groups compared to that in con?trol group. The increased catecholamines production from Th17 cells in lymph nodes may be involved in the anti-inflammato?ry progression in CIA.

Objective To summarize the clinical features of bee sting caused aute kidney injury in children and the effect of he-modialysis therapy .Methods 60 children cases of bee sting caused acute renal injury were performed the retrospective analysis on the clinical features and laboratory data .Results Cystatin C ,serum creatinine ,blood urea nitrogen in children patients with acute kidney injury after hemodialysis were significantly decreased ,while procalcitonin ,high-sensitivity C-reactive protein ,acidosis and e-lectrolyte imbalance also were corrected or improved .After hemodialysis in 60 children cases ,45 cases were clinically cured ,13 cases were significantly improved and discharged and 2 cases died .Conclusion Bee sting is most likely to result in children acute renal in-jury .Hemodialysis is safe and effective treatment measures for acute kidney injury ,can actively improve the renal function without significant complications of acid-base balance and electrolyte imbalance .But the prognosis is closely related with the onset age of the aute kidney injury ,injury severity ,where or not accompanied by MOF ,opportunity of diagnose and treatment ,etc .