Objective To prepare and identify rabbit polyclonal antibody against embryonic liver fordrin 3(ELF3),and investigate the distribution of ELF3 in mice tissue.Methods ELF3 specific N-terminal peptide was synthesized,and conjugated to Keyhole limpet hemocyanin(KLH)as immunogen.The ELF3-KLH complex was injected into rabbits subcutaneously,and then ELF3 antibody was purified using affinity chromatography.The titer of the antibody was evaluated by ELISA.The specificity of antibody against ELF3 and immunolocalization of ELF3 were evaluated by using Western blot and immunohistochemistry.Results Rabbit polyclonal antibody against ELF3 was prepared by the immunization of ELF3-KLH complex.ELISA and Western blot results showed the antibody against ELF3 had high titer and specificity.Western blot and immunohistochemical studies demonstrated ELF3 was expressed in the mouse heart,liver,brain and kidney tissue,particular on the cell membrane.Conclusion The preparation of polyclonal antibody against ELF3 was successful due to its high titer and specificity;ELF3 was expressed in the mice heart,liver,and kidney,particular on the cell membrane.It will provide an excellent tool for further study on the ELF3 function.

Objective:To discuss the diagnosis and treatment of urothelial carcinoma of the prostate.Methods:The clinical data of 25 patients with prostate urothelial carcinoma from January 2011 to November 2019 were retrospectively analyzed.Among the 25 patients, the age of onset was 39 to 85 years old, with an average of (63.4±11.2) years old, 13 patient presented with gross hematuria, 9 patients presented with dysuria, and 3 presented with bladder irritation. The PSA of 25 patients was within the normal range (less than 4 ng/ml). 17 cases of pelvic MRI showed abnormal signals in the bladder and prostate area, 3 cases indicated that prostate cancer had invaded the bladder, and 14 cases considered bladder cancer Invasion of the prostate suggests a cauliflower-like abnormal signal in the bladder area. 6 of this 14 patients have a history of bladder cancer. All 25 patients underwent surgical treatment, and 14 underwent transurethral diagnostic resection, of which 6 cases accepted radical cystectomy later. One patient underwent radical cystectomy combined with pelvic lymph node dissection 15 days after the first operation.9 cases received radical cystectomy.2 cases undergoing transurethral palliative resection due to multiple metastases before the operation.Results:The postoperative pathological diagnosis of 25 cases were all prostate urothelial carcinoma, 13 cases were accompanied by bladder urothelial carcinoma, secondary prostate urothelial carcinoma, and 12 cases were primary prostate urothelial carcinoma. After the operation, 13 patients were further treated. Among the patients with secondary prostate urothelial carcinoma, 7 patients received bladder perfusion, 2 patients received GC chemotherapy, 1 patient received local pelvic radiotherapy.25 patients were followed up for 2 to 36 months with an average of (21.5±10.1) months. Among them, lymph node metastasis were seen in 17/25 patients. lymph node metastasis were found in 7/25 before surgery, and 1/25 found lymph node metastasis during surgery. Among the patients with distant metastases afterwards, multiple metastases throughout the body (4/14), lung metastases only (6/14), and bone metastases only (4/14), the 1-year survival rate was 88% (22/25), the 2-year survival rate was 40% (10/25), and the 3-year survival rate was 28% (7/25).Conclusions:The diagnosis depends on histopathological examination. Early diagnosis may help improve prognosis. The first choice is a comprehensive treatment based on radical surgery.

To investigate the RNA interference (RNAi) effect induced by vector-derived small interfering RNA (siRNA) targeting the three gatekeeper genes (Rad52, Ku70, Ku80) and screen the more effective target sites from candidates for further research, by using siRNA design tools online, we selected 2 candidate sequences directed to every gatekeeper gene. According to the sequences, six vector-derived siRNAs (denoted psiRNA1-6) and one mocking psiRNA7 were constructed. Among them, psiRNA1 and psiRNA2 targeted Rad52, psiRNA3 and psiRNA4 to Ku70, psiRNA5 and psiRNA6 to Ku80. The mocking psiRNA7 was used as control. After sequence identification, the seven plasmids were transfected into HepG2 cell line. siRNA-induced silencing of gatekeeper genes was determined by using RT-PCR at RNA level and Western Blot at protein level. The results showed that the six plasmids specifically targeting the coding region of gatekeeper genes were successfully designed and constructed. To some extent, the six plasmids could reduce the expression of target gene. Comparatively, the plasmid-derived siRNA psiRNA1, psiRNA4 and psiRNA5 were more effective than their counterparts. The results suggest that the gene silencing efficiency of siRNA is different, depending on their targeted region, and siRNA may provide us with practical tools for further study on the three gatekeeper genes, i.e. Rad52, Ku70, Ku80.

Objective:To investigate the factors related to recurrence and prognosis of retroperitoneal liposarcoma.Method:The clinical data of patients with primary retroperitoneal liposarcoma who underwent surgical treatment in the First Affiliated Hospital of Zhengzhou University from June 2011 to January 2020 were analyzed retrospectively. There were 42 males and 47 females and patients’median age was 53 (26-78). Sixty-five cases were treated by operation in our hospital, and 24 cases were primarily treated by the operation in another hospital. The clinical manifestations of the initial diagnosis included retroperitoneal mass in 41 cases, abdominal distension in 12 cases, abdominal pain in 10 cases, fever in 11 cases, nausea, vomiting and poor appetite in 8 cases, frequent urination and dysuria in 6 cases, and bilateral lower limb edema in 1 case. Preoperative CT imaging showed that the tumor body was located in the retroperitoneal kidney area in 58 cases, while in the retroperitoneal space or the pelvic extraperitoneal space in 31 cases. There were 55 single cases and 34 multiple cases. The median tumor length was 20(3-52) cm. Among the primarily treated 65 patients, 47(72.3%) were considered as primary retroperitoneal liposarcoma by preoperative imaging examination. Among the 89 patients treated by surgery, 78 underwent endoscopic surgery, among which 21 underwent laparoscopic surgery, 38 cases of retroperitoneal laparoscopic surgery, 19 cases of Da Vinci robot-assisted laparoscopic surgery. Open operation was performed in 11 cases. There were 87 patients undergoing radical resection and 2 patients undergoing palliative resection. Forty-two patients underwent intraoperative combined resection of the adjacent organs. The recurrence and survival status of patients were followed up.Results:All the 89 patients underwent the operation successfully, with the median operative blood loss of 200 (10-2000) ml. There were 23 cases being diagnosed of well differentiated liposarcoma, 40 cases of dedifferentiated, 20 cases of myxoid/round, 5 cases of myxoid liposarcoma, and 1 cases of mixed type. Pathologically, there 42 cases with low grade histology and 47 cases with high grade histology. In this study, 89 patients were followed up for 3 to 108 months, and the median follow-up time was 28 months. The 5-year recurrence free survival rate, disease-free survival rate and overall survival rate of the patients were 16.7%, 16.1% and 52.6%, respectively. There were 57 patients presenting local recurrence, 1 patient of lung metastasis, and 1 patient of liver metastasis, and the median disease-free survival time was 24 months. There were 42 patients died of the disease, with a median survival time of 64 months. Univariate analysis showed that intraoperative blood loss( P<0.01), whether multiple cases( P<0.01), pathologic types( P<0.01), and histological grades ( P<0.01) were related to disease-free survival.The intraoperative blood loss( P<0.01), multiple cases( P<0.05), pathologic types ( P<0.05), and recurrence ( P<0.01)were related with overall survival. Gender, age, tumor size, tumor location, whether primary surgery, radical resection or combined resection of adjacent organ had no effect on the prognosis of patients ( P>0.05). Cox regression model multivariate analysis revealed that surgical bleeding ( RR=2.360, 95% CI 1.313-4.241, P=0.004), multiple tumor ( RR=1.899, 95% CI 1.068-3.375, P=0.029), and pathological type ( RR=4.976, 95% CI 1.622-15.264, P=0.005) were independent factors affecting disease-free survival. The recurrence was an independent factor affecting the overall survival of patients ( RR=31.495, 95% CI 1.062-933.684, P=0.046). Conclusions:Retroperitoneal liposarcoma is a rare disease with high recurrence rate. The intraoperative blood loss, whether multiplicity and pathological type are independent factors affecting the disease-free survival, and recurrence is independent factors affecting the overall survival.

Objective:To explore the clinicopathological characteristics and prognostic factors of bladder squamous cell carcinoma (SqCC)and bladder adenocarcinoma.Methods:A retrospective analysis of the clinical data of 107 patients with nonurothelial carcinoma of the bladder admitted to the First Affiliated Hospital of Zhengzhou University from October 2011 to January 2019 was performed. Among the 107 patients, 78 were males and 29 were females(ratio 2.69∶1), and the median age of onset was 62.0 years. According to histological types, patients were divided into SqCC group, urachal adenocarcinoma group and non-urachal adenocarcinoma group. There were 55 cases in the SqCC group, including 40 males and 15 females. Their mean age was 69.0(58.0, 75.0) years. 14 cases had the history of smoking. The clinic manifestation included hematuria in 35 cases, bladder irritation in 13 cases, dysuria in 2 cases and pain in 5 cases .Tumors located at the anterior and posterior walls in 18 cases, at the lateral wall in 27 cases, at the triangular area in 8 cases and at the apical wall in 2 cases. The average diameter of tumor was 4.5(3.0, 6.0) cm. 37 cases suffered with single tumor and 18 cases suffered with multiple tumors. The surgical method was radical cystectomy in 38 cases, partial cystectomy in 4 cases, TURBT in 9 cases, interventional surgery in 2 cases, and no operation in 2 cases. There were 20 cases in the urachal adenocarcinoma group, including 14 males and 6 females; age 53.5(43.5, 57.8) years; 6 cases with a history of smoking. The clinic manifestation included hematuria in 16 cases, bladder irritation in 1 case, pain in 2 cases and asymptomatic in 1 case. Tumors located at anterior and posterior walls in 4 cases and at apical wall in 16. The average diameter of tumor was 3.0(2.0, 4.8) cm. Single tumor was present in 18 cases, multiple tumors were present in 2 cases. The surgical method was partial cystectomy in 16 cases, radical cystectomy in 1 case, TURBT in 1 case and no operation in 2 cases. There were 32 cases in the non-urachal adenocarcinoma group, including 24 males and 8 females. Their mean age was 55.0(46.3, 70.8) years.11 cases had a history of smoking. The clinic manifestation included hematuria in 16 cases, bladder irritation in 3 cases, dysuria in one case and pain in 7 cases. Tumor located at anterior and posterior walls in 17 cases, at lateral wall in 7 cases, at triangular area in 5 cases and at apical wall in 3 cases. The average diameter of tumor was 3.6(2.0, 4.5) cm. 23 cases suffered with single tumor, 9 cases suffered with multiple tumors.The surgical method was radical cystectomy in 11 cases, partial cystectomy in 9 cases, TURBT in 9 cases , and no operation in 3 cases. The preoperative data of the three groups of tissue types were compared, the differences of age of onset, tumor diameter, tumor location, reason for treatment, operation method ( P<0.05)among the 3 groups were statistically significant. The clinicopathological characteristics and prognosis of the 3 groups of tissue types were compared, and the Cox proportional regression risk model was used to analyze the clinical factors affecting the prognosis. Results:91 patients were followed up, the overall follow-up rate was 85.1%, the median follow-up time was 26(7, 48) months. The 3-year and 5-year overall survival rates were 54.1% and 42.2%, respectively. In the SqCC group, 11 cases received chemotherapy; 3 cases received postoperative radiotherapy; 12 cases received postoperative perfusion.10 cases had recurrence; 17 cases had lymph node metastasis; 19 cases had distant metastasis; 5 cases were pT x in pT stage, 36 cases were pT 1-pT 2, 14 cases were pT 3-pT 4. 19 cases had unknown tumor differentiation, 4 cases had well differentiated, 24 cases had moderately differentiated and 8 cases had poorly differentiated. In the urachal adenocarcinoma group, 7 cases received chemotherapy, 3 cases had recurrence, 2 cases had lymph node metastasis, 2 cases had distant metastasis; 1 case was pT x in pT stage, 16 cases were pT 1-pT 2, 3 cases were pT 3-pT 4. 9 cases had unknown tumor differentiation, 3 cases had well differentiated, and 5 cases had moderately differentiated, 3 cases had poorly differentiated. In the urachal adenocarcinoma group, 3 cases received chemotherapy, 1 case received postradiotherapy, 11 cases received postoperative perfusion; 10 cases had recurrence. 5 cases had lymph node metastasis, 4 cases had distant metastasis, 6 cases were pT x, 21 cases were pT 1-pT 2 and 5 cases were pT 3-pT 4. 14 cases had unknown tumor differentiation, 8 cases had moderately differentiated and 10 cases had poorly differentiated.The postoperative general data of the three groups of tissue types were compared. There was statistically significant difference between whether there was postoperative perfusion and whether there was distant metastasis (all P<0.05). The univariate analysis results showed gender, age, surgical method, lymph node metastasis, distant metastasis, pT staging, tumor differentiation and histological type were risk factors that affect the prognosis(all P<0.05). Cox multivariate analysis showed that women ( HR=2.604, P=0.032) and distant metastases ( HR=2.571, P=0.026) were independent risk factors affecting the prognosis of patients. Conclusions:SqCC and adenocarcinoma are clinically rare and have poor prognosis. They often present with hematuria. Surgical treatment is the first choice. Radical cystectomy is the first choice for SqCC and non-urachal adenocarcinoma, and extended partial cystectomy is the first choice for urachal adenocarcinoma. Female and distant metastasis are independent risk factors affecting the prognosis of patients.

OBJECTIVETo assess the association of rs9272346 polymorphism of HLA-DQA1 gene with clinical outcome of hepatitis B virus (HBV) infection in ethnic Han population from Hubei, China.

METHODSA case-control study was conducted, which have involved 1028 unrelated subjects including 238 asymptomatic HBV carriers (AHC), 173 acute liver failure (ALF), 292 liver cirrhosis (LC) and 325 hepatocellular carcinoma (HCC). Genotypes of rs9272346 were determined by real-time polymerase chain reaction with a TaqMan MGB probe. Statistical results were analyzed using Chi square test, student's t test and unconditional logistic regression.

RESULTSNo significant differences were detected in the frequencies of G allele between ALF, LC, HCC and AHC groups (P= 0.312, 0.314, 0.264). Compared with the AA genotype, the GG and GA genotypes were not associated with the patients groups under the dominant model: ALF group vs. AHC group (adjusted OR= 1.08, 95%CI: 0.7-1.68), LC group vs. AHC group (adjusted OR= 1.11, 95%CI: 0.87-1.26), HCC group vs. AHC group (adjusted OR= 0.93, 95%CI: 0.65-1.33). For women, the GG and GA genotypes have conferred a protective effect for the outcome of ALF (OR= 0.30, 95%CI: 0.1-1.87).

CONCLUSIONOur results suggested that rs9272346 polymorphism of HLA-DQA1 may not independently influence the outcome of HBV infection in ethnic Han Chinese in Hubei, while the GG and GA genotypes may confer a protective effect against ALF in women.

Adult ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; Female ; Genetic Predisposition to Disease ; Genotype ; HLA-DQ alpha-Chains ; genetics ; Hepatitis B ; genetics ; Humans ; Male ; Middle Aged ; Polymorphism, Genetic

Objective To investigate the clinical features and treatment principles of inflammatory myofibroblastic tumor of the urinary bladder (IMTUB).Methods From April 2013 to October 2016,6 cases of IMTUB patients were analyzed retrospectively.All cases were presented with gross hematuria.4 cases underwent ultrasonography,of which 3 cases showed solid mass in bladder,1 case showed inflammatory change.6 cases underwent CT examination,3 cases with bladder cancer,1 case with bladder sarcoma,1 case with malignant transformation of adenoma,1 case with rich blood supply.No lymph node metastasis.Bladder occupying lesions were considered in 2 cases of MRI examination.5 cases of cystoscopy showed bladder solid mass.In 6 cases involved,2 patients received partial cystectomy,2 patients underwent transurethral resection of bladder tumor,1 patient underwent radical resection of urachal carcinoma and the other one was treated with chemotherapy.Results Immunohistochemical staining was positive in ALK (100.0％) 、Vimentin(100.0％) 、CK(100.0％) 、SMA (83.3％) 、EMA(66.7％) and Ki-67 (5％-30％),negative in S-100 and Desmin.Final pathological diagnosis was IMTUB.So far,neither recurrence nor metastasis has been detected for 6 ～ 42 months in 5 cases and the other one lost to follow-up.Conclusions IMTUB is a kind of rare benign tumor of bladder.The golden standard of diagnosis is pathological diagnosis.Surgical resection is the first choice for treatment.Recurrence and metastasis are after the surgery treatment.All patients should be followed up closely.

Objective:To analyze the diagnosis and treatment of adrenal lymphangioma.Methods:From September 2011 to March 2019, 16 case of adrenal lymphangioma (8 males, 8 females), aged from 15 to 65 years were retrospectively analyzed. Eleven cases were discovered accidentally by physical examination or imaging examination when treating other diseases, with 2 patients presenting lumbago, 2 patients presenting abdominal pain and 1 presenting weakness. CT showed that the tumors were cystic or cystic solid low density mass without enhancement (9 cases in the right, 7 cases in the left) and some had calcified lesions. The maximum diameter of the tumors was 2.5-16.0 cm. The endocrine examination was abnormal in 6 patients, including 2 patients with elevated 24-hour urinary vanillic mandelic acid (VMA) and adrenaline, 1 patient with elevated 24-hour urinary free cortisol (UFC) and elevated blood cortisol, and 3 patients with increased aldosterone. They were considered sub-clinical adrenal tumors. All patients were monitored for blood pressure, and patients with elevated VMA were given terazosin for volume expansion; Patients with elevated UFC are given hydrocortisone 200 mg intravenously during operation, hormone dosage is adjusted in time according to blood cortisol levels, and patients with elevated aldosterone were monitored on blood aldosterone and potassium. All cases underwent surgery, including 10 cases of laparoscopic adrenalectomy, 3 cases of laparoscopic adrenal tumor resection, 1 case of open adrenalectomy, and 2 cases of open adrenal tumor resection. One of the open surgery was an emergency exploratory laparotomy for ruptured lymphangioma.Results:The operations were successful, and there was no operation related complications.Sixteen cases were diagnosed as adrenal lymphangioma. The gross specimens were monocystic or multilocular cyst, and the cyst could be detected in the section. HE staining showed that the mass was composed of irregular cystic cavity filled with lymphoid fluid. A single layer of flattened endothelium lines the walls of lymphatic channels. Immunohistochemical staining was positive in D2-40(7/7), CD34(6/6), CD31(6/6), SMA (3/3) and CR(2/2); negative in CK(7/7) and CK7(3/3). So far, no recurrence has been detected after 6-95 months follow-up.Conclusions:Adrenal lymphangioma is a rare benign lesion clinically, without typical clinical symptoms. Preoperative diagnosis mainly depends on imaging examination. It is a non-functional lesion, whose endocrine examinations are mostly normal. Histopathology is reliable diagnostic modality. The therapy can refer to the principle of treatment for adrenal incidental tumors. Surgery is the first choice and prognosis is good.

Host genetic, environmental and viral factors are classified as three categories that determine clinical outcomes of hepatitis B virus (HBV) infection. The objective of this study was to detect the associations between polymorphisms rs346473 and rs346482 in Rho GTPase-activating protein 24 (ARHGAP24) gene and disease progression of HBV infection in Han Chinese population. These two SNPs were found by our DNA pooling using Affymetrix Genome-Wide Human Mapping SNP6.0 Array in HBV carriers, and verified by using TaqMan 7900HT Sequence Detection System with 758 progressed HBV carriers versus 300 asymptomatic HBV carriers (AsC) in a discovery phase and 971 progressed HBV carriers versus 328 AsC in a replication phase. Multivariable logistic regression revealed that individuals with genotype TT at variant rs346473 displayed remarkable correlations with disease progression of HBV infection both in the discovery phase (OR, 2.693; 95% CI, 1.928-3.760; P=6.2×10(-9); additive model) and the replication phase (OR, 1.490; 95% CI, 1.104-2.012; P=9.0×10(-3); additive model). These two SNPs were in strong linkage disequilibrium with D'=0.99 and r (2)=0.951, and haplotype TT disclosed an increased susceptibility to HBV progression (OR, 1.980; 95% CI, 1.538-2.545; P=8.1×10(-8)). These findings suggest that polymorphism rs346473 in the ARHGAP24 gene might be a part of the genetic variants underlying the susceptibility of HBV carriers to disease progression.

To investigate the RNA interference (RNAi) effect induced by vector-derived small interfering RNA (siRNA) targeting the three gatekeeper genes (Rad52, Ku70, Ku80) and screen the more effective target sites from candidates for further research, by using siRNA design tools online,we selected 2 candidate sequences directed to every gatekeeper gene. According to the sequences, six vector-derived siRNAs (denoted psiRNA1-6) and one mocking psiRNA7 were constructed. Among them, psiRNA1 and psiRNA2 targeted Rad52, psiRNA3 and psiRNA4 to Ku70, psiRNA5 and psiRNA6 to Ku80. The mocking psiRNA7 was used as control. After sequence identification, the seven plasmids were transfected into HepG2 cell line. siRNA-induced silencing of gatekeeper genes was determined by using RT-PCR at RNA level and Western Blot at protein level. The results showed that the six plasmids specifically targeting the coding region of gatekeeper genes were successfully designed and constructed. To some extent, the six plasmids could reduce the expression of target gene.Comparatively, the plasmid-derived siRNA psiRNA1, psiRNA4 and psiRNA5 were more effective than their counterparts. The results suggest that the gene silencing efficiency of siRNA is different,depending on their targeted region, and siRNA may provide us with practical tools for further study on the three gatekeeper genes, i.e. Rad52, Ku70, Ku80.