ObjectiveTo explore the inhibitory effects of r-k4k5 on retinal neovascularization.MethodsEighty-eight one-week-old C57BL/6J mice were put into the environment with 75% oxygen for 5 days to establish models of vascular proliferation retinopathy. One eye of each mouse received an intravitreal injection of 500 ng of r-k4k5 (large-dosage group) and of 250 ng of r-k4k5(small-dosage group), and the same volume of BSS was injected into the other eye of the mice both in these two groups as a control. The ADPase histochemical staining was used for retinal flatmount to observe changes of retinal vessels. The inhibitory effects of r-k4k5 on retinal neovascularization were evaluated by counting the endotheliocyte nuclei of new vessels extending from retina to vitreous in the tissue-slice.ResultsRegular distributions and reduced density of retinal blood vessels in eyes in the treatment group were found in retinal flatmount. The number of the endotheliocyte nuclei of new vessels extending from retina to vitreous was less in the eyes in the treatment group than which in control group (P

Objective To evaluate the efficacy and safety of 131I-metaiodobenzylguanidine (131 I-MIBG) in treatment of malignant pheochromocytoma/malignant paraganglioma (MPHEO/MPGL).Methods The clinical data of 96 cases of MPHEO/MPGL (60/36) treated with 131I-MIBG between December 1998 and April 2014 were retrospectively reviewed.Among them,the malignant pheochromocytoma was found in 60 cases and malignant paraganglioma was found in 36 cases.Seventy-eight patients (81.2％)presented initially with hypertension,whereas 18 patients (18.8％) presented adrenal incidentaloma.Before 131I-MIBG treatment,24 h urinary norepinephrine was (409.5± 127.2) nmol,24 h urinary dopamine was (99.3±41.1) nmol,24 h urine adrenaline was (1 409.9±336.0) nmol.Before treatment,the compound iodine solution was given to each one.Then,all patients were given an initial course of 131I-MIBG therapy (5.55,7.40 GBq).Subsequent 131I-MIBG treatment (5.55,7.40 GBq) was undertaken every three to six months.The patients got symptomatic,hormonal or radiological response underwent sbsequent 131I-MIBG therapy (3.70,5.55 GBq) every year.All patients underwent clinical symptoms (headache,palpitate,sweating,hypertension),biochemical (24 h urine catecholamin) and radiological evaluation (CT/MRI) within 6 months to evaluate the efficacy and safety of 131I-MIBG treatment.Results After one to eleven sessions of 131 I-MIBG treatment,in total,266 doses of 131 I-MIBG were administered,average dose was 6.49 GBq.22.9％ of patients demonstrated radiological partial response (≥ 50％ reduction in tumor size) after first or repeated 131 I-MIBG treatment.Eleven cases (11.5％) achieved clinical complete response,41 cases (42.7％) achieved clinical partial response and 23 cases (24.0％) maintained the stable clinic symptoms.After treatment,24 h urinary norepinephrine (164.3±71.6) nmol and dopamine (49.7±24.7) nmol showed significantly decline,compared with those before treatment (P＜ 0.05).While,24 h urinary epinephrine (1 354.7±433.4) nmol had no obvious change (P＞0.05).No life-threatening adverse events were reported,but 2 MPGL patients developed transient leucopenia or thrombocytopenia after four and five times 131 I-MIBG treatment,respectively.Conclusions Treatment with repeated low dose 131I-MIBG is well tolerated and effective in controlling the progression and alleviating the clinical symptoms.The 131I-MIBG therapy is an effective and safe treatment modality for MPHEO/MPGL.

Objective To investigate the clinical features, diagnosis and treatment of cardiac pheochromocytoma. Methods The clinical data of 3 cases (2 males and 1 female) who were 17,40 and 35 years old,respectively,were analyzed.They all presented with headache, sweating, palpitations and hypertension.Before operation 24-hour urinary catecholamine excretion was examined for the 17-year-old man and the woman twice with the results as follows.In the 17-year-old man,epinephrine (E) was 1.84,150.28 ?g/24h (normal range,4.08?2.34 ?g/24h);norepinephrine (NE):734.77,508.00 ?g/24h (normal range,28.67?11.98 ?g/24h);dopamine (DA):823.00,620.31 ?g/24h (normal range,225.76?104.83 ?g/24h).In the woman,E was 12.40,16.03 ?g/24h;NE:1778.92,1243.96 ?g/24h;DA:1517.47,926.89 ?g/24h.All the 3 patients had positive result with Tc-99m-Otretide scintigraphy,and all underwent orthotopic excision of tumors under cardiopulmonary bypass. Results During operation,the tumors of 2-5 cm in size were seen to lie at the base of the aorta,left atrium and interauricular septa respectively,and grew invasively like carpet,thus were difficult to remove. Postoperative pathology of 3 cases confirmed the diagnosis of cardiac pheochromocytoma.After follow-up for 8-24 months,3 cases had a mean fall of 30 mmHg in systolic pressure and 20 mmHg in diastolic pressure,respectively.The 17-year-old man had decline in cardiac function that was proved by Tc-99m-Otretide scintigraphy test;another male patient suffered from arrhythmias 1 month after operation,and was found to have tumor remains by PET test.Myocardial infarction occurred in the female patient. Conclusions Cardiac pheochromocytoma is a rare tumor, and its diagnosis is rather difficult and complex.Surgical excision is very difficult with the possibility of complications.

Objective:Cancer stem cells (CSCs) are resistant to chemotherapy. Our study aimed to investigate the stem cell-like proper-ties of doxorubicin-resistant osteosarcoma cell line 143B and its correlation with Notch signaling. Methods:We generated doxorubicin-resistant osteosarcoma cells by treating them with 2μm doxorubicin. Stem cell-like properties such as morphology change, Stro-1/CD117 double positive ratio, stem cell-related gene expression, sphere formation efficiency, and EMT character were assessed on day 5 after doxorubicin withdrawal. Notch receptor and its target genes were examined using qPCR and Western blot analysis. The stem cell-like properties of doxorubicin-resistant osteosarcoma cells were assessed when pretreated with Notch inhibitor or vehicle. The an-ti-tumor effect of Notch inhibitor was tested using a xenograft model. Results:Doxorubicin-resistant osteosarcoma cells were enriched in Stro-1+/CD117+cells, which showed obvious increased expression of stem cell-related genes, and exhibited enhanced spheroid for-mation and evident mesenchymal characteristics unlike doxorubicin-sensitive cells. qPCR and Western blot assays showed that Notch intracellular domain 1 (NICD1) and target genes Hes1 and Hey1 were upregulated in doxorubicin-resistant osteosarcoma stem cells compared with those in vehicle cells. Furthermore, pretreatment with a γ-secretase inhibitor (GSI) to prevent Notch signaling en-hanced chemo-sensitivity and inhibited doxorubicin-enriched osteosarcoma stem cell activity in vitro. Finally, the Notch inhibitor pre-vented tumor growth in mice xenograft models. Conclusion: Doxorubicin induced the enrichment of osteosarcoma stem-like cells through Notch signaling, and inactivation of Notch could be useful for overcoming drug resistance and eliminating osteosarcoma.

Objective To elucidate gene mutation and promoter methylation changes of p16 gene in pheochromocytomas (PHEO) and paragangliomas (PGL) and to assess its relation with tumor clinical characters. Methods A total of 34 tumors (20 PHEO, 14 PG L, 15 benign, 19 malignant) were collected.Direct sequencing of p16 gene after PCR was performed to analyze genetic alterations. Hypermethylation of p16 gene promoter CpG island was analyzed by methylation specific PCR(MSP). In addition, mRNA expression was detected by RT-PCR. Results Homozygous deletion and gene mutation were not observed in 34 PHEO and PGL. Aberrant methylation of p16 gene promoter CpG island was found in 35.3% (12 of 34 tumors, 3 PHEO, 9 PGL). The p16 promoter hypermethylation in PGL was significantly higher than PHEO (P=0. 005). The higher p16 promoter hypermethylation was associated with malignant behavior, tumor number, and younger age at presentation, but no statistical significance, due to the limited number of cases. The p16 mRNA expression in malignant cases was lower than in benign tumors(0.83±0.65 vs 1.12±0.81 ,P=0.278). Conclusion p16 gene homozygous deletion and mutation were not frequent in PHEO and PGL. The promoter hypermethylation is mainly attributed to inactivation of the p16 gene.

Objective To analys hyperinsulinemia in Bartter syndrome. Methods Twenty-three cases of Bartter syndrome [age (27 ±9) years;fasting serum potassium(2. 8 ±0. 5)mmol/L], 20 patients of aldosterone-producing adenoma [APA, age (45 ± 11 ) years, fasting serum potassium ( 3.0 ± 0. 4 ) mmol/L], 20 patients of idiopathic hyperaldosteronism [IHA, age (51 ± 11 ) years, fasting serum potassium (3.4 ±0. 2)mmol/L] were diagnosed in Peking Union Medical College Hospital from September 2003 to May 2008. All patients underwent 3-hours oral glucose tolerance test(3hOGTT), postural stimulation test and calculated HOMA-insulin resistance ( HOMA-IR ) and HOMA-insulin sensitivity ( HOMA-IS ) by Homeostasis model.Results The insulin area under curve-(229.0±162.4)mIU·L-1·h] was singnificantly higher than APA group [(227.7±158.6)mIU·-1·h].But HOMA-IR in Bartter group were similar to APA group( 1.96 ± 1.14 vs 1.41 ± 0. 91 ), and HOMA-IR in APA group was lower than IHA group ( 1.96 ± 1.14 vs 2.40 ± 1.60, P ＜ 0. 05 ). There was no deference in HOMA-IS among three groups,but APA group had lower level. In all three groups, the peak of insulin secretion was delayed. Conclusion Bartter syndrome patients commonly present with hyperinsulinemia.

Objective To investigate the value of X-ray guided desmopressin (DDAVP) stimulated bilateral inferior petrosal sinus sampling (IPSS) in diagnosing the recurrence of ACTH dependent Cushing disease or ineffectiveness after surgery or radiotherapy. Methods Retrospective analyses of patients with recurrent ACTH dependent Cushing disease (31 cases) or ineffective (3 cases) treatment after surgery or radiotherapy from January 2013 to October 2014 in our hospital was conducted. Bilateral inferior petrosal sinus angiography showed the same side of the cavernous sinus to prove successful intubation. The cases with discontinuous of the inferior petrosal sinus and cavernous sinus were excluded by this study. Finally, there were 34 cases of the patients in this study. Diagnosis was based on the ratio of ACTH level in IPS to peripheral vein after desmopressin test.The gradient≥2 at baseline or gradient≥3 after desmopressin test suggested the sources were in the pituitary. Diagnosis was confirmed by gold standard to investigate the value of X-ray guided desmopressin (DDAVP) stimulated bilateral IPSS. Results The IPS gradient≥2 at baseline or gradient≥3 after desmopressin test suggested the sources were in the pituitary in 30 patients. A total of 22 (22/30) patients underwent surgery with a final diagnosis of ACTH adenoma. The symptoms were obviously relieved in 8 (8/30) cases after sellar area gamma knife treatment and lesions were confirmed in the pituitary. IPS gradient<2 at baseline or gradient<3 after desmopressin test was found in 4 cases. One case (1/4) was found to have for ACTH adenoma after pituitary surgery. The other 3 cases (3/4) were confirmed to have lung carcinoid and clinical symptom alleviated after surgery. The sensitivity of desmopressin stimulated IPSS was 96.8%, the specificity was 100%, and the accuracy was 97.1%. Conclusion Desmopressin stimulated IPSS is an effective diagnostic procedure in diagnosing ACTH dependent Cushing disease recurrence or ineffectiveness after surgery or radiotherapy.

Objective To investigate the role of miR-429 in osteosarcoma stem cells.Methods We employed different approaches to test the expression of miR-429 in osteosarcoma stem cells.miR-429-high and miR-429-low cells were separated using molecular probes and their stem cell-like properties were compared.The effect of miR-429 on osteosarcoma stem cell-like properties was further tested through transfection assays.Furthermore,the miR-429 downstream target gene was confirmed by luciferase assay.Results The expression of miR-429 in osteosarcoma stem cells was much lower than that in control cells.miR-429-high cells displayed fewer stem celllike properties than did miR-429-low cells.These observations were confirmed by transfection assays.Additionally,our luciferase assays showed that miR-429 regulates Sox2 at the post-transcriptional level.Conclusion miR-429 negatively regulates osteosarcoma stem celllike properties by targeting Sox2.

Objective:To compare the protective effect of corneal protectant hydroxypropyl methylcellulose (HPMC) and balanced salt solution (BSS) on corneal epithelium in vitreous body surgery for proliferative diabetic retinopathy (PDR).Methods:A randomized controlled clinical trial was conducted.Fifty eyes of 50 patients with PDR who underwent minimally invasive vitrectomy in Xuzhou First People's Hospital from September 2018 to January 2019 were enrolled and randomly divided into HPMC group and BSS group, with 25 eyes in each group.All eyes underwent three-channel minimally invasive vitrectomy under non-contact wide-angle lens, and 2% HPMC and BSS were applied to keep the ocular surface moist according to grouping during the operation.The combination of cataract surgery, operation time, application times of 2% HPMC or BSS and the grade of corneal epithelial edema before and after operation were recorded.The patients were followed up for 1 month.Corneal fluorescein sodium staining and tear film break-up time (BUT) test were performed before operation and at 1 day, 3 days, 10 days and 1 month after operation.Schirmer Ⅰ test (SⅠt) and ocular surface disease index (OSDI) questionnaire were carried out before operation and at 1 month after operation.The study protocol adhered to the Declaration of Helsinki and was approved by an Ethics Committee of Xuzhou First People's Hospital (No.xyyⅡ[2018]005). Written informed consent was obtained from each patient prior to any medical examination.Results:The average frequency of application of eye drops in the HPMC group was (5.00±1.56) times, which was significantly less than (50.56±17.00) times in the BSS group ( t=-13.071, P<0.01). The postoperative corneal edema score of the HPMC group was 1.0 (1.0, 1.0) point, which was lower than 2.0 (2.0, 2.0) points of BSS group, and the difference was statistically significant ( Z=-4.909, P<0.01). There were statistically significant differences in postoperative corneal fluorescein sodium scores among 1 day, 3 days, 10 days and 1 month in the two groups (HPMC group: χ2=36.040, P<0.01; BSS group: χ2=50.892, P<0.01). The 1-, 3- and 10-day postoperative corneal fluorescein sodium scores in the HPMC group were significantly lower than those in the BSS group (all at P<0.05). One-day, 3-day, 10-day and 1-month postoperative BUT values were (6.15±2.20), (6.95±2.46), (6.16±2.11) and (5.81±2.92) seconds in the HPMC group, respectively, and were (3.89±1.87), (5.32±2.59), (5.01±2.12) and (4.97±2.10) seconds in the BSS group, respectively.There were significant differences in BUT between the two groups at different time points ( Fgroup=5.240, P<0.05; Ftime=2.846, P<0.05). The preoperative and 1-month postoperative SⅠt values in the BSS group were (12.24±5.55)mm and (9.96±4.53)mm, respectively, showing a statistically significant difference ( t=3.863, P=0.001). The postoperative OSDI score in the BSS group was (51.00±12.04) points, which was significantly higherly than the preoperative value of (47.89±10.95) points, and the difference was statistically significant ( t=-2.111, P<0.05). Conclusions:Compared with BSS, 2% HPMC in minimally invasive vitreous surgery has better protective effect on corneal epithelium of PDR, reduces the frequency of eye drop use during the surgery and shortens the repair time of ocular surface tissue after surgery.

Objective:To compare the efficacy, recurrence rate and safety of a single intravitreal injection of ranibizumab between type 1 prethreshold and threshold retinopathy of prematurity (ROP).Methods:A retrospective cohort study was conducted.A total 100 eyes from 51 ROP patients in Xuzhou Municipal Hospital Affiliated to Xuzhou Medical University from January 2016 to December 2019 were enrolled.Retcam and indirect ophthalmoscope were used to confirm the diagnosis of 39 cases of type 1 prethreshold ROP (70 eyes) and 18 cases of threshold ROP (30 eyes). All the subjects were given a single intravitreal injection of ranibizumab (IVR) 0.25 mg/0.025 ml as the initial treatment within 24-36 hours after diagnosis.Events such as increased intraocular pressure, intraocular bleeding and endophthalmitis were observed and recorded 1 day after IVR.Retcam photography was used to check whether the fundus lesions was subsided 1 week after IVR.Regular follow-up was conducted once every 2-4 weeks until retinal vascularization.Systemic and ocular complications were recorded.The cure rate, the time of retinal vascularization and the recurrence rate of single IVR for type 1 prethreshold disease and threshold disease were compared.This study was approved by the Ethics Committee of Xuzhou Municipal Hospital Affiliated to Xuzhou Medical University.Results:After a single IVR, 86 of 100 eyes (86.00%) were cured, among which 60 of 70 type 1 prethreshold ROP eyes (85.71%) and 26 of 30 threshold ROP eyes (86.67%) were cured, with no significant difference between the two groups ( χ2=0.09, P=0.65). No systemic adverse reactions were observed after a single IVR treatment.The final retinal vascularization time was (11.86±8.60) weeks in the type 1 prethreshold ROP group and (14.45±10.01) weeks in the threshold ROP group, with no statistically significant difference between the two groups ( t=0.88, P=0.22). Recurrence of ROP appeared in 10 eyes after a single IVR, including 4 eyes (5.71%) in the type 1 prethreshold ROP group and 6 eyes (20.00%) in the threshold ROP group, the recurrent rate was significantly different between the two groups ( χ2=4.76, P=0.03). Six recurrent eyes were treated with IVR for the second time, and 4 recurrent eyes were treated with laser photocoagulation.Five eyes presented with preretinal hemorrhage after IVR treatment, and were recovered without intervention. Conclusions:The treatment of type 1 prethreshold ROP and threshold ROP with single IVR in vitreous cavity is safe and effective.The recurrence rate of threshold ROP is higher than type 1 prethreshold ROP.Retreatment can significantly improve the cure rate.