OBJECTIVE: To report the clinical and genetic characteristics of a rare case of Gitelman syndrome with comorbid Graves disease and ACTH-independent adrenocortical adenoma. METHODS: A patient who had presented at the Nanchong Central Hospital on December 21, 2020 was selected as the study subject. Clinical data of the patient was collected. Whole-exome sequencing was carried out on DNA extracted from peripheral venous blood samples from the patient and her family members. RESULTS: The patient, a 45-year-old woman, was found to have Graves disease, ACTH-independent Cushing syndrome, hypokalemia and hypomagnesemia following the discovery of an adrenal incidentaloma. MRI scan had revealed a 3.8 cm × 3.2 cm mass in the left adrenal gland. The mass was removed by surgery and confirmed as adrenocortical adenoma. DNA sequencing revealed that the patient and her sister have both harbored compound heterozygous variants of the SLC12A3 gene, namely c.1444-10(IVS11)G>A and c.179(exon1)C>T (p.T60M), which were respectively inherited from their father and mother. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the c.1444-10(IVS11)G>A and c.179(exon1)C>T (p.T60M) were respectively classified as a variant of uncertain significance (PM2_Supporting+PP3) and a likely pathogenic variant (PM3_Strong+PM1+PP3). CONCLUSION The conjunction of Gitelman syndrome with Graves disease and adrenal cortex adenoma is rather rare. The newly discovered c.1444-10(IVS11)G>A variant of the SLC12A3 gene, together with the heterozygous variant of c.179(exon1)C>T (p.T60M), probably underlay the pathogenesis in this patient.
Humans ; Female ; Middle Aged ; Gitelman Syndrome/genetics* ; Adrenocortical Adenoma ; Hypokalemia ; Graves Disease/genetics* ; Mothers ; Mutation ; Solute Carrier Family 12, Member 3

Objective:To investigate whether Losartan can inhibit the formation of ascites in a nude mouse model of ovarian cancer by improving lymphatic drainage.Methods:Eight-week-old nude mice were randomly divided into 3 groups with 24 mice in each group.The blank control group received no treatment.Mice in the other two groups were given orthotopic transplantation of SKOV3 IP1-Gluc human ovarian cancer cells.Peritoneal tumor growth was monitored by peripheral blood Gluc levels, and mice were enrolled when the blood Gluc level reached 1×10 6 relative light units.Twenty-two nude mice were in the control group and 23 in the Losartan treatment group.Mice were sacrificed on the 28 th day after cancer cell implantation.Tumor tissues and diaphragm were removed, and the volume of ascites was measured after suction.Sirius Red, αSMA and Collagen 1 staining were used to detect the contents of extracellular matrix collagen and fibroblasts.Lymphangiography was used to examine the diaphragmatic lymphatic morphology, and intraperitoneal injection of fluorescent microspheres was used to visualize the distribution of fluorescent microspheres in diaphragmatic and mediastinal lymph nodes to estimate the function of diaphragmatic lymphatic drainage. Results:Losartan treatment significantly reduced the formation of ascites in the nude mouse model of ovarian cancer( P<0.05). Intratumoral contents of collagen(sirius red positive area, Collagen 1 content)and fibroblasts(αSMA positive area)were significantly reduced in the Losartan treatment group(all P<0.05). Compared with normal nude mice, the diaphragm lymphatic vessels were dilated and the structure was disordered in the control group.In the Losartan treatment group, the morphology and distribution of lymphatic vessels in the diaphragm improved and tended to normalize, as did the drainage function of lymphatic vessels. Conclusions:Losartan can inhibit the formation of extracellular matrix, reduce the extracellular matrix content of ovarian cancer cells, reduce solid pressure, relieve the oppressive effect on lymphatic vessels, improve lymphatic drainage and reduce ascites formation.

Objective: To analyze the antimicrobial resistance profile in Chinese children. Methods: This was a prevalence survey. From January 1 through December 31, 2016, the isolates were collected from 10 tertiary children hospitals in China. Antimicrobial susceptibility testing was carried out by routine laboratory methods. The penicillin susceptibility of streptococcus pneumonia and Meropenem susceptibility of gram-negative bacteria were detected by E-test and disk diffusion method respectively. Antimicrobial susceptibility results were interpreted according to the criteria of Clinical and Laboratory Standards Institute (CLSI) Guideline 2016. The data of antimicrobial susceptibility testing of isolates from either the different patients (neonatal group and non-neonatal group) or various sources were analyzed by WHONET 5.6 software. Results: A total of 56 241 isolates were collected, of which 41.5% (23 328 isolates) were gram-positive organisms and 58.5% (32 886 isolates) gram-negative organisms. The five leading pathogens were Escherichia coli (7 995/56 214, 14.2%), Straphylococcus aureus (6 468/56 214, 11.5%), Streptococcus pneumonia (6 225/56 214, 11.1%), Haemophilus influenza (5 435/56 214, 9.7%) and Klebsiella pneumonia (4 523/56 214, 8.0%). The Meropenem resistance rates of Klebsiella pneumonia, Enterobacter cloacae, Escherichia coil, Pseudomonas aeruginosa, Acinetobacter baumonia isolates were 27.4% (326/1 189) , 8.1% (29/358) , 2.0% (27/1 362) , 19.5% (34/174) , 49.7% (230/463) in neonatal group and 15.4% (512/3 327) , 4.8% (40/841) , 2.3% (151/6 564) , 13.7% (252/1 840) , and 53.4% (860/1 611) in non-neonatal group. The Methicillin-resistant Staphylococcus aureus (MRSA) rates of neonatal group and non-neonatal group were 46.2% (649/1 404) and 33.3% (1 668/5 010) . The penicillin non-susceptible rates of Streptococcus pneumonia in the two groups were 17.6% (6/34) and 18.2% (1 121/6 158) respectively. The β-lactamase positive rates of Haemophilus pneumonia isolates in the neonatal group and non-neonatal groups were 33.8% (47/139) and 44.4% (2 345/5 282) respectively. Conclusion This investigation highlights the worrisome trend of antimicrobial resistance in children, especially among neonatal patients in China.

Objective To investigate the expressions of matrix metalloproteinases(MMPs) in Kashin-Beck disease(KBD) cartilage as well as in a KBD rat model of T-2 toxin poisoning under selenium deficient conditions, and to investigate the effect of T-2 toxin on MMP-13 expression in human chondrocytes in vitro in order to determine a possible mechanism underlying KBD. Methods Samples of articular cartilage were divided into 2 groups:controls(samples from 5 normal children, traffic accident or operation), and KBD(samples from 5 children with KBD, auctopsy). Thirty-two Sprague-Dawley rats were divided into two groups by body weight using random number table: normal diet group(n = 16) and selenium-deficient diet group(n=16). The selenium level in normal diet was 101.500μg/kg, and in selenium-deficient diet was 1.118μg/kg. Rats were fed for 4 weeks with selenium-deficient or normal diet, respectively. After successful build up of the low selenium rat model, normal diet group was then subdivided into 2 sub-groups: normal group(n = 8) and normal diet plus low T-2 toxin group(n = 8);and selenium-deficient diet group was also subdivided into 2 sub-groups: selenium-deficient group ( n = 8 ) and selenium-deficient diet plus T-2 toxin group ( n = 8 ) . T-2 toxin of 100 μg·kg-1·d-1 was administered by intragastric administration for 30 days. Then the rats were sacrificed, and their knee joints were processed for histopathological evaluation. MMP-1 and MMP-13 locations in cartilages were performed by inmmunohistochemistry. Human chondrocytes C28/I2 were cultured in vitro. The experiment was divided into 4 groups: empty vector plasmid group, MMP-13 promoter plasmid group, MMP-13 promoter plasmid plus 20 μg/L T-2 toxin group and MMP-13 promoter plasmid plus 40 μg/L T-2 toxin group. MMP-13-luciferase reporter plasmid and vector plasmid were transiently transfected into C28/I2 cells for 24 hours, and then treated with 20 - 40 μg/L T-2 toxin for 24 hours. Transactivation of human MMP-13 promoter was analyzed using luciferase reporter constructs containing sequences spanning-1602 to+20 bp in C28/I2 chondrocytes. Results The percentages of chondrocytes staining for MMP-1 in the superficial and middle zones of KBD samples [(29.73 ± 10.12)%, (28.27 ± 0.91)%] were significantly higher than those of controls[(2.47 ± 0.11)%, (0.00 ± 0.00)%, all P < 0.05]. The percentages of chondrocytes staining for MMP-13 in the superficial and middle zones of KBD samples [(13.21 ± 4.32)%, (41.85 ± 6.32)%] were significantly higher than those of controls[(5.72 ± 0.31)%, (0.00 ± 0.00)%, all P<0.05]. The percentages of chondrocytes staining for MMP-13 in the superficial and middle zones of rats fed with selenium-deficient diet plus T-2 toxin group[(13.21 ± 4.32)%, (61.85 ± 8.68)%] were significantly higher than those of the normal and selenium-deficient groups[(2.43 ± 0.22)%, (5.89 ± 0.69)%, (3.03 ± 0.29)%, (25.99 ± 0.57)%, all P < 0.05]. Moreover, T-2 toxin activated the MMP-13 promoter detected with luciferase reporter assays in C28/I2 cells. The luciferase activities in MMP-13 promoter plasmid plus 20 μg/L T-2 toxin group and MMP-13 promoter plasmid plus 40μg/L T-2 toxin group(0.082 78 ± 0.008 40, 0.103 35 ± 0.013 19) were significantly higher than those in empty vector plasmid group and MMP-13 promoter plasmid group(0.024 19 ± 0.000 96, 0.040 32 ± 0.003 56, all P < 0.05). Conclusions These data suggest that T-2 toxin induces cartilage matrix degradation through up-regulation of MMP-13 promoter expression. Increased MMPs staining intensity in KBD cartilage and the rat KBD model of T-2 toxin poisoning under selenium deficient conditions suggest that matrix degradation appear to be driven by MMPs activity.

Objective To investigate the expressions of interleukin-1β(IL-1β), interleukin-6(IL-6) and tumor necrosis factor alpha(TNF-α) in cartilage of children with Kashin-Beck disease(KBD) in order to provide a possible mechanism of the disease. Methods Articular cartilage tissues of 5 KBD children(KBD group) were selected from KBD children autopsy samples keeping in Institute of Endemic Diseases, Medical School of Xi’an Jiaotong University; articular cartilage tissues of 5 normal children ( control group ) were selected from non-KBD areas of Shaanxi Province, three cases were from accident death children, two cases were the samples of congential malformation of six finger. Expressions of IL-1β, IL-6 and TNF-α in the cartilage were detected using immunohistochemistry; the cells of articular cartilage were divided into three areas (superficial zone, middle zone and deep zone) to analyze the expressions of IL-1β, IL-6 and TNF-α. Results The expressions of IL-1β in superficial zone , middle zone and deep zone of articular cartilage of KBD group (63.50 ± 7.19, 54.75 ± 5.50, 66.20 ± 9.91) were significantly higher than those of control group(5.75 ± 1.26, 0.00 ± 0.00, 0.00 ± 0.00, all P<0.05). The expression of IL-6 in superficial zone of articular cartilage in KBD group(55.25 ± 6.24) was significantly higher than that of control group(0.00 ± 0.00, P<0.05). The expressions of TNF-αin all zone of articular cartilage of KBD group(33.25 ± 6.50, 3.75 ± 0.96, 29.80 ± 1.92) were significantly higher than those of control group (3.74 ± 0.82, 0.00 ± 0.00, 0.00 ± 0.00, all P < 0.05). Conclusion The levels of IL-1β, IL-6 and TNF-α are up-regulated in articular cartilage of KBD children, suggesting that cytokines may play an important role in matrix degradation in KBD children cartilage.

BACKGROUNDGiant cell interstitial pneumonia (GIP) was a rare form of pneumoconiosis, associated with exposure to hard metals, which had been reported mostly as isolated case reports. We described eight cases of GIP diagnosed in our hospital during the past seven years, with particular reference to new findings.

METHODSEight patients with GIP confirmed by biopsy in the Nanjing Drum Tower Hospital affiliated to Medical School of Nanjing University from 2005 to 2011 were retrospectively analyzed. For each patient, the occupy histories and medical records were thoroughly reviewed and clinic data were extracted. Two radiologists, without knowledge of any of the clinical and functional findings, independently reviewed the HRCT scans of all patients. Follow-up data were collected.

RESULTSAmong the eight patients, seven had a history of exposure to hard metal dusts, one denied an exposure history. The most common manifestations were cough and dyspnea. One patient initiated with pneumothorax and another pleural effusion, both of which were uncommon to GIP. The main pathologic appearances were the presence of macrophages and multinucleated giant cells in the alveolar space. The clinical symptoms and radiographic abnormalities were obviously improved after cessation of exposure and receiving corticosteroid treatments, recurrences were observed in two patients when they resumed work. In spite of exposure cessation and corticosteroid treatment, one patient developed pulmonary fibrosis at seven years follow-up.

CONCLUSIONSAwareness of the patients' occupational history often provided clues to the diagnosis of GIP. Histopathologic examinations were necessary to establish the right diagnosis. Exposure cessation was of benefit to most patients; however, pulmonary fibrosis was possible in spite of exposure cessation and corticosteroid treatment. Better ways should be found out to improve the outcome and quality of life.

Adult ; Alloys ; toxicity ; Cobalt ; toxicity ; Female ; Humans ; Lung ; pathology ; Lung Diseases, Interstitial ; diagnosis ; etiology ; Male ; Middle Aged ; Pulmonary Fibrosis ; diagnosis ; etiology ; Retrospective Studies ; Tungsten ; toxicity ; Young Adult

Objective To examine matrix proteoglycan metabolic markers and probe into the turnover of matrix proteoglycan and enzyme-mediated role of matrix metalloproteinases (MMPs) and aggrecanases in reparative tissues with tissue engineering cartilage. Methods Tissue-engineered cartilage was constructed by cancellous bone matrix gelatin (BMG) with allogeneic chondrocytes in vitro for 2 weeks, then implanted to repair osteochondral defects of rabbit knee joint. Samples were obtained 6 months later to explore the expressions of 3-B-3(-) epitope, MMPs, MMP-generated epitope BC-4 and aggrecanases-generated epitope BC-13. Results In repaired tissues, the expression of 3-B-3(-) epitope increased, but that of MMPs and MMP-generated epitope BC-4 reduced. There was no expression of aggrecanases-generated epitope BC-13. Conclusion Expressions of 3-B-3(-), MMPs, BC-4 and BC-13 can help probe into the matrix proteoglycan turnover in reparative cartilage tissues. Anabolism exceeds catabolism in the repaired tissues. MMPs play an important role in the conservative baseline turnover of proteoglycan and remodeling of the graft tissues.

Objective To analyze the clinical presentations and radiological characteristics of acute exacerbation of idiopathic pulmonary fibrosis (IPF).Methods Clinical and radiological data of 2 patients with acute exacerbation of IPF from April 2006 to July 2008 were retrospectively analyzed and literatures were reviewed.Results Both patients were senior male patients over 60 years old.Dyspnea, cough and inspiratory crackles were the major symptoms and signs.Two patients were experiencing an exacerbation of dyspnea for one week and half of month,respectively.PaO2/FiO2 of both patients was less than 225 mm Hg.In both patients, high-resolution computed tomography (HRCT) scans at the exacerbation showed typical signs of IPF including peripheral predominant, basal predominant reticular abnormality, with honeycombing and traction bronchiectasis and bronchiolectasis,and newly developing alveolar opacity.HRCT scan showed peripheral area of ground-glass attenuation adjacent to subpleural honeycombing in one patient,and diffusely distributed ground-glass opacity in another patient.Two patients had received cortieosteroid treatment.For one patient, the symptoms improved, and ground-glass attenuation adjacent to subpleural honeycombing had almostly resolved.The other patient died of respiratory failure.Conclusions Some acute exacerbation in idiopatic pulmonary fibrosis can be idiopathic.The chnical presentations mainly include the worsening of dyspnea within short time.HRCT generally demonstrates new bilateral ground-glass abnormality with or without areas of consolidation, superimposed on typical changes of IPF.

Objective To construct shifting mutant of cysteine-rich region to 3?@terminal of Tat gene of human immunodeficiency virus type-1 (HIV-1) HXB2 strain, and to analyze the immunogenicity of mutant protein (Tat-cct) after prokaryotically expressed and purified. Methods The cysteine-rich region (nucleotides 64--111) of Tat gene was shifted to 3'terminal of Tat of HIV-1 HXB2 strain by polymerase chain reaction (PCR) and Tat mutant DNA sequence was obtained. Prokaryotie express plasmid pET32a-Tat-cct was constructed and transformed into E. coli BL21 (DE3), then Tat-cct protein was expressed and purified. BALB/c mice were immunized with the fusion protein Tat-cct, and immunogenicity of the immunized serum was detected by enzyme-linked immunosorbent assay (ELISA). Results The recombinant plasmid pET32a-Tat-cct expressed in E. coli BL21 (DE3) and the relative molecular mass of the purified fusion protein was 31 000. The serum antibody titer of mice immunized with Tat-cct recombinant protein was 1 : 1600, which binded specifically with both Tat-ect protein and Tat protein (amino acids 1-101). Conclusions The recombinant protein Tat-cct of Tat mutant strain can be expressed efficiently in E. coli and well retains immunogenicity, which provides valuable information for basic research of HIV-1 Tat vaccine.

Objective To explore the value of hysteroscopy in diagnostic curettage, the incidence of complications and preventive measures in senile women. Methods One hundred and thirteen senile patients who underwent hysteroscopy from January 2002 to December 2007 were recruited in the study, and one hundred and fifty-seven non senile patients were selected as control group. All the patients were operated with diagnostic curettage and hysteroseopy. The success rate of hysteroscopy operation, the incidence of complications and control effect were recorded and compared between two groups. Results The success rate of hysteroscopy operation was 88. 5% (100/113) in the senile group and 100% (157/157) in the non senile group, and there was significant differenee(P <0. 01). The incidence of complications was higher in the senile women group than that in the non senile women group (P< 0. 01). In senile group, the accuracy of diagnostic curettage and hysteroscopy was higher than diagnostic curettage alone, and the success rate was 95.5% (64/67)in estrogen group and 69.6%(32/46) in blank control group (P<0. 01) ,which indicated that estrogen was positively related with the success rate of operation. Compared with misoprostol, estradiol-pretreatment operation was safer and had higher success rate. Conclusions It is safe and feasible to perform hysteroscopy and diagnostic curettage for older women, though it has higher incidence of complications and lower success rate compared with non senile women. Hysteroscopy and diagnostic curettage are more accurate than diagnostic curettage alone, and it is a good choice to apply estradiol before operation.