Objective To investigate the feasibihty and effectiveness of the fluorescence in situ hybridization(FISH)technique in the diagnosis of renal pelvic tumor.Methods FISH technique were used to detect the abnormalities of chromosome 3,7,17 and 9 p16 from 20 people without malignancy.The morning's first urine were availahled from 15 patients with hematuria and suspected of renal mass by imaging,then were studied by FISH technique and urine cytology respectively.The sample was considered positive if two or more probes results were higher than the criteria,or one probe had two or more abnormal results.Results Eight of 15 patients with hematuria were renal pelvic urothehal carcinoma,5 were renal clear cell carcinoma,1 was renal chromophobe cell carcinoma,1 was xanthogranulomatous pyelonephritis,confirmed by pathology.All of the 8 renal pelvic carcinoma were considered positive by FISH technique,while only 1 was considered positive by urine cytology.The variation rate of chromosome 3,7,17 were 100%(8/8)and chromosome 9 p16 was 75%(6/8).Six of 7 non-renal pelvic carcinoma were negative by FISH technique,but all Were negative by urine cytology.Conclusion FISH technique is a non-invasive and effective method for the diagnosis of renal pelvic tumor,and it's more sensitive than urine cytology.

Objective To discuss the diagnosis and treatment of Cushing's disease without remission after transsphenoidal surgery.Methods 12 patients of Cushing's disease without remission after transsphenoidal surgery were investigated in this retrospective study.There were 2 males and 10 females with an average age of 36 (21-46) years old.Cushing's disease was diagnosed with the typical symptoms and signs of Cushing's syndrome,endocrinological examination and CT or MRI.Transsphenoidal surgery was performed in all patients.None of the patients achieved remission after operation.Serum and urinary free cortisol (UFC)were still elevated 3 months after operation.Subtotal adrenalectomy combined with pituitary radiotherapy were performed in 5 patients,single subtotal adrenalectomy in 4,pituitary radiotherapy in 2 and bilateral adrenalectomy with adrenal autotransplantation in 1 patient.Results Those receiving subtotal adrenalectomy combined with pituitary radiotherapy had no recurrence or Nelson's syndrome during 20 years of follow-up,but 4 patients receiving single subtotal adrenalectomy recurred after 2 to 9 years after operation.2 patients achieved remission with pituitary radiotherapy 1 to 2 years later without recurrence during 5 years of follow-up.One patient who underwent bilateral adrenalectomy with adrenal autotransplantation needed steroid replacement and Nelson's syndrome occurred during 2 years of follow-up because transplanted adrenal gland had no function.Conclusion The diagnosis of Cushing's disease should be reconfirmed for patients without remission after transsphenoidal surgery and the treatment should be individualized based on the results of operation,pathology and imaging.

The contents of cholesterol in a number of vegetable oils were determined by saponification-gas chromatography ( FID) . There was quite a large possibility that cholesterol peak was seriously interfered by saponification-gas chromatography ( FID) , so it was difficult to set a cholesterol content value to differentiate vegetable oil from waste oil. Solid phase extraction ( SPE) sample pretreatment was chosen and the process conditions were optimized. The optimal conditions were as follows:0. 25 g of oil samples, 20 mL of 0. 6%ethyl ether-hexane ( V/V) as eluent ( get rid of fatty acid glycerides) and 20 mL of 15% ethyl ether-hexane ( V/V) as eluent ( obtain cholesterol) . An obvious cholesterol-content-difference between vegetable oils and waste oils was found by SPE-GC. The detected cholesterol contents in 84 vegetable oil samples were all less than 50 μg/g and the contents of 11 waste oil samples among the 13 waste oils were greater than 50 μg/g. Therefore, cholesterol content in vegetable oil of more than 50 μg/g could be determined as suspected waste oil, not vice versa by SPE-GC. The correlation coefficient R2 was 0. 9999 in 0-760 mg/L concentration range, and the detection limit was 6. 0 μg/g. Relative standard deviations at two concentration levels of 17. 7 and 695 mg/L were 1. 6% and 1. 5% respectively. The recovery was 103%.

Objective To discuss the diagnosis and treatment of primary adrenal lymphoma(PAL).Methods Two cases of PAL were retrospectively analyzed and the literature was reviewed.Results One patient received laparoscopic right adrenalectomy and was diagnosed as B cell Non Hodgkin's lymphoma by pathology.Immunohistochemical stains showed positive for CD20 (B cell originmarker) but negative for CD3.0ne cycle CHOP ( eyclophosphamide,doxorubicin,vincristine,andprednisone) chemotherapy was given post operation but the patient died three months after operationbecause of dyscrasia.Another patient received 2-stage bilateral laparoscopic adrenalectomy.Immuno histoehemical staining demonstrated positive CD3 (T cell origin marker) activity,but negative CD20 ac tivity and T cell Non l lodgkin's lymphoma was confirmed.Then the patient received four cyclesCHOP chemotherapy and was in good condition during S months follow up.Conclusions PAL is arare disease and has a poor prognosis.Most reported PAL patients who received only one therapeuticmodality have unsatisfactory survival rates.A combination of therapeutic modalities such as surgeryfollowed by chemotherapy and/or radiotherapy may improve prognosis for patients with PAL than sin gle modality therapy.

Objective To evaluate the value of using fluorescence in situ hybridization (FISH)technique for the detection of chromosome aberration of urine exfoliated cells for the diagnosis of bladder tumor.MethodsFISH technique were used to detect the abnormalities of chromosome 3,7,17 and 9p16 site from 20 normal people, and to establish the threshold.The morning's first urinations were available from 75 patients with bladder cancer and 25 patients without urothelial tumor, then were detected using FISH technique and urine cytology respectively.The sample was considered positive if two or more probes results higher than the criteria,or one probe has two or more abnormal results.Results The sensitivity of single using were 73.3％ (55/75),76.0％ (57/75),62.7％ (47/75) and 62.7％ (47/75) for the 4 probes (3,7, 17 and 9p16)respectively.The sensitivity of combined detection was 85.3％ (64/75) and specificity was 96.0％ (24/25) The sensitivity and specificity of urine cytology examination was 9.3％ (7/75) and 100％ (25/25) .The sensitivity of FISH examination was significantly higher than that of urine cytology examination (85.3％ vs 9.3％ ,x2 = 57.00, P ＜ 0.001) .Sensitivity of FISH examination was not correlated with cancer pathologic grading(low vs high : 84.2％ vs 86.5％, x2 = 0.08, P ＞ 0.05)and clinical stage (ta-tl : 82.9％, t2-t4 :87.5％, x2 = 0.32 ,P ＞ 0.05) .ConclusionFISH technique is a non-invasive and effective method for the early diagnosis of bladder tumor and is more sensitive than urine cytology.Furthermore, FISH technique can be used to predict the tumor's biological behavivor and prognosis.

Objective To discuss the clinical feasibility and significance of retroperitoneal laparoscopic adrenalectomy for bilateral macronodular adrenal hyperplasia.Methods 9 patients with bilateral macronodular adrenal hyperplasia received retroperitoneal laparoscopic operations.5 cases had the typical clinical manif(e)stetions of Cushing's syndrome and 4 patients presented hypertension or diabetes mellitus without any sign of Cushing's syndrome.Body mass index was 18.8-31.2,with 25.0 as the average.Serum cortisol was 544-1746 nmol/L,with 984 nmol/L as the average.Urinary free cortisol (UFC) was 347-1989 nmol/24h with 1129 nmol/24h as the average.CT scan showed bilateral enlargement of the adrenal glands with multiple macronodules.The size of the adrenal gland was 6 cm × 5 cm × 4 cm to 10 cm × 6 cm× 3 cm,with 8 cm× 5 cm × 4 cm as the average.Results One patient underwent retroperitoneal laparoscopic bilateral adrenalectomy with replantation of adrenal gland tissue in scrotum,1patient underwent right lateral adrenalectomy combined with left subtotal adrenalectomy,4 patients underwent right lateral adrenalectomy ( 1case was converted to open surgery due to hemorrhage),and 3 patients underwent left lateral adrenalectomy.The average operation duration was 116 min,ranging from 60 to 165 min.The estimated average intraoperative blood loss was 110 ml,ranging from 20 to 600 ml.No severe complications like injury in great vessels or organs happened during the operation.Pathological diagnosis showed nodular or adenomatoid hyperplasia of adrenal cortex.The average postoperative follow-up was 58 months,ranging from 12 to 84 months.Clinical symptoms of Cushing's syndrome disappeared after surgery in the 5 cases with typical clinical features of Cushing's syndrome.Body weight and blood glucose recovered to normal level for those 4 cases presenting body weight increasement,hypertension or diabetes mellitus.2 cases with right lateral adrenalectomy received left adrenalectomy 33 and 41months after the first operation due to recurrence of Cushing's syndrome.Conclusions Retroperitoneal laparoscopic adrenalectomy for bilateral macronodular adrenal hyperplasia is technically feasible in experienced hands. Unilateral adrenalectomy can relieve most of the Cushing's symptoms.Bilateral adrenalectomy or unilateral adrenalectomy combined with contralateral subtotal adrenalectomy can be performed if the symptoms are not improved or recurred after unilateral adrenalectomy.

Objective To explore the diagnosis and treatment of adrenocorticotropin-independent macronodular adrenal hyperplasia (AIMAH). Methods The clinical symptoms, endocrinological and radiological characteristics, treatment modalities, and postoperative clinical course of 12 AIMAH patients were retrospectively reviewed. There were 5 men and 7 women with an average age of 48 years old. Results All cases had some clinical features and biochemical evidence of Cushing's syndrome. The circadian rhythm of serum cortisol was abnormal. Low- and high-dose dexamethasone suppression tests failed to suppress cortisol secretion. CT or MRI scan showed bilateral enlargement of the adrenal glands with multiple macronodules. Bilateral adrenalectomy was performed in three patients and unilateral adrenalectomy combined with contralateral subtotal adrenalectomy was performed in one patient. Seven patients underwent unilateral adrenalectomy of the larger gland. The removed adrenal glands weighted within 8-96 g range and pathologic diagnoses were nodular or adenomatoid hyperplasia of adrenal cortex. The duration of postoperative follow-up was 14 to 124 months. Clinical symptoms of Cushing's syndrome disappeared after surgery in all cases and there was no Nelson's syndrome after bilateral adrenalectomy. Urinary free cortisol and serum cortisol were within normal ranges. No further enlargement of the contralateral gland has been noticed in patients who received unilateral adrenalectomy during the follow-up.Conclusions AIMAH had unique endocrinological and pathological features. Unilateral adrenalectomy appears to be an effective and safe alternative treatment for AIMAH. Bilateral adrenalectomy or unilateral adrenalectomy combined with contralateral subtotal adrenalectomy may be performed if the symptoms had not been improved or recurred after unilateral adrenalectomy.

OBJECTIVETo detect potential mutation in a Chinese family affected with succinic semialdehyde dehydrogenase deficiency.

METHODSGenomic DNA was extracted from the peripheral blood samples of the proband, her family members and 100 healthy controls. All exons and flanking intronic regions of the ALDH5A1 gene were amplified by PCR and subjected to direct sequencing.

RESULTSThe proband was found to have compound heterozygous mutations of the ALDH5A1 gene, namely c.398_399delAA (p.N134X) and c.638G>T (p.R213L), for which her parents were both heterozygous carriers. The same mutations were not found among the 100 healthy controls.

CONCLUSIONThe novel mutations of the ALDH5A1 gene probably underlie the pathogenesis of the disease in the infant, which also enriched the mutation spectrum of the ALDH5A1 gene.

Amino Acid Metabolism, Inborn Errors ; ethnology ; genetics ; Amino Acid Sequence ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; DNA Mutational Analysis ; methods ; Developmental Disabilities ; ethnology ; genetics ; Exons ; genetics ; Family Health ; Female ; Heterozygote ; Humans ; Infant ; Introns ; genetics ; Male ; Mutation ; Sequence Homology, Amino Acid ; Succinate-Semialdehyde Dehydrogenase ; deficiency ; genetics

OBJECTIVETo investigate α-toxin-induced apoptosis of umbilical vein endothelial cells and explore its role in vertical infection of Staphylococcus aureus L-form.

METHODSHUV-EC-C cells exposed to different concentrations (0, 10, 30, 90, and 270 ng/ml) of α-toxin for different time lengths (0, 2, 4, 6, and 8 h) were examined for apoptosis using flow cytometry with Annexin V-PI staining. The levels of tumor necrosis factor-α (TNF-α) and the activities of, caspase-3 and caspase-8 in the cell culture were detected by ELISA and colorimetric method, respectively. α-Toxin-induced cell apoptosis was also analyzed in HUV-EC-C cells treated with a neutralizing antibody of TNF-α or with the inhibitory peptides of caspase-3 (zDEVD-FMK) and caspase-8 (zIETD-fmk).

RESULTSα-Toxin induced apoptosis of HUV-EC-C cells in a dose- and time-dependent manner and caused significantly enhanced expression of TNF-α and the activation of both caspase-3 and caspase-8. Inhibition of TNF-α with its neutralizing antibody and the inhibitory peptides of caspase-3 or -8 all significantly decreased α-toxin-induced cell apoptosis, and the caspase-3 inhibitor completely blocked α-toxin-induced cell apoptosis.

CONCLUSIONα-Toxin-induced apoptosis is partially mediated by the extrinsic cell death pathway of TNF-α and caspase-8 and plays an important role in the vertical infection of S. aureus L-form to affect fetal growth and development.

Apoptosis ; Bacterial Toxins ; toxicity ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Cells, Cultured ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; L Forms ; Staphylococcal Infections ; Staphylococcus aureus ; Tumor Necrosis Factor-alpha ; metabolism

Abstract: Heterologous boost COVID-19 vaccination can solved the problem of decreased efficacy caused by single dose of vaccine. Heterologous booster with adenoviral-vectored COVID-19 vaccine (Ad5-nCoV) following primary immunization with inactivated COVID-19 vaccines is a widely-used vaccination strategy in clinic, while different routes of Ad5-nCoV administration exist and pose a question which route could be more optimal. In this study, we comprehensively evaluated and compared the vaccine immunity induced in mice immunized according to three different vaccination regimens: “3×phosphate buffered solution(3× PBS)”, “2×inactivated vaccine + 1×inactivated vaccine (3×INA)”, “2×inactivated vaccine + 1×Ad5-nCoV (intramuscular)[2×INA+Ad5(im)]”and“2×inactivated vaccine + 1×Ad5-nCoV (intranasal)[2×INA+Ad5(in)]”. We found that heterologous booster with Ad5-nCoV, irrespective of the route of administration, induced significantly higher levels of anti-Spike IgG and subclasses (IgG1and IgG2c), Spike-specific T cells, class-switched Spike+ memory B cells (MBCs) than homologous booster with 3rd dose of inactivated COVID-19 vaccine. Of note, compared with the intramuscular given, intranasal given of Ad5-nCoV as a booster dose clearly induced higher levels of serum and bronchoalveolar bavage fluid anti-spike immunoglobulin A, and moreover, induced stronger infiltration of major innate effector cells like neutrophils, natural killer cells and dendritic cells into the lung tissue, which suggested that mucosal vaccine responses are generated upon intranasal booster with Ad5-nCoV. Altogether, our study analyzed the vaccine immunity induced by different COVID-19 vaccines administered using different regimens, which may guide the clinical use of other types of prophylactic vaccines aiming to mount improved vaccine responses.