[Clinical and StAR genetic characteristics of 33 children with congenital lipoid adrenal hyperplasia].

Objective: To analyze the clinical and genetic characteristics of 33 children with congenital lipoid adrenal hyperplasia (CLAH) caused by StAR gene defects. Methods: The clinical, biochemical, genetic, and follow-up (until December 2021) data of 33 children diagnosed with CLAH from 2006 to 2021 were retrospectively analyzed in Xinhua Hospital, Shanghai Jiao Tong University School of Medicine. Results: Of the 33 children with CLAH, 17 had a karyotype of 46, XX and 16 had a karyotype of 46, XY; 31 were female and 2 were male by social gender. Classic type and non-classic type were found in 30 and 3 children respectively. The age at diagnosis was 9.0 (3.0, 34.5) months. All the 30 cases with classic CLAH presented within the first year of life with skin hyperpigmentation (28 cases, 93%), vomiting and(or) diarrhea (19 cases, 63%), no increase in body weight (8 cases, 27%), elevated adrenocorticotropic hormone levels (21cases (70%)>275 pmol/L), decreased cortisol levels (47 (31,126) nmol/L), hyponatremia ((126±13) mmol/L), hyperkalemia ((5.7±1.1) mmol/L), and normal 17α-hydroxyprogesterone levels (30 cases, 100%). All these with classic CLAH exhibited female external genitalia. Three children with non-classic CLAH (including 2 cases of 46, XY and 1 case of 46, XX) also showed signs and symptoms of adrenal insufficiency, but 2 of them had an age of onset later than 1 year of age, including 1 case of 46, XY with male external genitalia and 1 case of 46, XX with female external genitalia. The other 46, XY patient with non-classic CLAH presented with adrenal insufficiency at 2 months of age, showing micropenis and hypospadias. In the 17 females with 46, XX, 4 older than 10 years of age showed spontaneous pubertal development. A total of 25 StAR gene pathogenic variants were identified in 33 patients, with p.Q258* (18/66, 27%), p.K236Tfs*47 (8/66, 12%) and p.Q77* (6/66, 9%) being the common variantion. Six novel variants were found, including c.358T>G, c.713_714del, c.125del, c.745-1G>A, c.179-2A>C, and exon 1 deletion. Conclusions: Patients with classic CLAH typically present with signs and symptoms of primary adrenal insufficiency in the early infancy period and female external genitalia. p.Q258*, p.K236Tfs*47 and p.Q77* are common variants in CLAH patients.

[Identification and Characterization of circRNAs in the Developing Stem Cambium of Poplar Seedlings].

Non-coding RNAs are a class of RNAs with multiple roles in plant life. Covalently closed circular RNA molecules (circRNAs) have been recently shown to be a group of RNA isoforms that show widespread tissue-specific expression in plants, often cooperating with the corresponding linear mRNAs to regulate gene function. However, no previous study of poplar has identified circRNAs in the cambium and determined their potential roles in the cambium or xylem development. In the present study, we sequenced RNAs in the cambium of poplar seedlings at two developmental stages, and identified and characterized 4912 circRNAs. Alternative back-splicing circularization events for 87 genes were identified among the circRNAs derived from different chromosomes. A total of 1138 circRNAs originated from 928 host genes, which were classified among the three major functional categories by GO analysis. Thirty-nine circRNAs were differentially expressed between cambium samples of stems at two developmental stages. Twenty-four DEcircRNAs interacted with 98 miRNAs as targets, of which some were associated with cambium growth and development. The results suggest that circRNAs play important roles in the cambium in relation to the regulation of stem growth and development in poplar seedlings.

[Application value of urinary IGFBP7 and TIMP-2 in acute kidney injury with decompensated hepatitis B virus-related liver cirrhosis].

Objective: To investigate the application value of new urinary biomarkers insulin-like growth factor binding protein 7 (IGFBP7) and tissue matrix metalloproteinase inhibitor-2 (TIMP-2) in acute kidney injury with decompensated hepatitis B virus-related liver cirrhosis. Methods: 45 newly hospitalized cases with decompensated hepatitis B virus-related liver cirrhosis were selected. Among them, 19 cases were combined with AKI on admission (cirrhosis-AKI group), 26 cases without AKI (cirrhosis-non-AKI group), and 12 healthy cases (normal control group). First-morning urine samples were collected and IGFBP7 and TIMP-2 were detected by enzyme-linked immunosorbent assay (ELISA). Urinary IGFBP7 and serum creatinine (SCr) were dynamically monitored after hospitalization in cirrhosis-non-AKI group. Normally distributed measurement data were compared by t-test, and non-normally distributed measurement data were compared by rank sum test. The receiver operating characteristic (ROC) curve and area under the curve (AUC) were used to evaluate the diagnostic accuracy of the indicators. Results: Urinary IGFBP7, IGFBP7 with TIMP-2 (IGFBP7×TIMP-2) in cirrhosis-AKI group (n = 19) were equally higher than that of the cirrhosis-non-AKI group (P < 0.05). Urinary IGFBP7, TIMP-2 and IGFBP7×TIMP-2 in cirrhosis-AKI group or cirrhosis-non-AKI group were significantly higher than those of the normal control group (P < 0.01). The AUC of urinary IGFBP7 and urinary IGFBP7×TIMP-2 for diagnosis of AKI were 0.703 (95% CI 0.547-0.860) and 0.700 (95% CI 0.541-0.859), respectively. In the liver cirrhosis-non-AKI group (n = 26), 5 cases of AKI were newly diagnosed according to the changes in SCr during hospitalization (progressive group). Urinary IGFBP7 was significantly increased 2 days before the diagnosis of AKI. The concentration of urinary IGFBP7 at admission in the progressive group (n = 5) was higher than that of the non-progressive group (n = 21) (P < 0.05). Conclusion: Urinary IGFBP7 and TIMP-2 concentrations were significantly increased in patients with decompensated hepatitis B virus-related liver cirrhosis. When AKI occurred, urinary IGFBP7 and IGFBP7×TIMP-2 was further increased. Urinary IGFBP7 is valuable for early AKI diagnosis, and may play a role in predicting AKI occurrence.

Effects of lncRNA MALAT1-mediated ß-catenin signaling pathway on myocardial cell apoptosis in rats with myocardial ischemia/reperfusion injury.

OBJECTIVE: To investigate the effects of long non-coding ribonucleic acid (lncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) on myocardial ischemia/reperfusion (I/R) injury in rats and its mechanism, and to provide a certain reference for the clinical prevention and treatment of myocardial infarction. MATERIALS AND METHODS: A total of 60 male Sprague-Dawley rats were divided into 3 groups using a random number table, including the Sham group (n=20), I/R group (n=20) and I/R + MALAT1 small interfering RNA (siRNA) group (n=20). An I/R model was established by means of recanalization after ligation of the left anterior descending coronary artery of the rats. The rats in the I/R + MALAT1 siRNA group were used to establish a model of MALAT1 knockdown by injecting MALAT1 siRNA from the tail vein. The myocardial infarction area in each group was detected via 2,3,5-triphenyl tetrazolium chloride (TTC) staining. The ejection fraction% (EF%) and fractional shortening% (FS%) of the heart in each group were measured through echocardiography. Hematoxylin and eosin (H&E) staining was adopted to determine the morphological changes in myocardial cells in each group. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining was performed to detect the apoptosis levels of myocardial cells and fibroblasts in the cardiac tissues in each group, and Western blotting assay was conducted to measure the expression levels of apoptosis-related proteins [B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax)]. In addition, the content of ß-catenin in the three groups of rats was determined via immunohistochemical staining. Finally, the impacts of MALAT1 siRNA on the expression level of ß-catenin protein were detected using Western blotting assay. RESULTS: MALAT1 siRNA could prominently ameliorate the I/R-induced cardiac insufficiency in the rats and improve the EF% and FS% of the heart (p<0.05). Moreover, MALAT1 siRNA was able to remarkably inhibit the I/R injury-induced myocardial infarction, reducing the infarction area from (59.54±3.45) to (24.85±1.30; p<0.05). The results of the H&E staining indicated that compared with those in the I/R group, the myofilaments of the myocardial cells were well-arranged, the degrees of degradation and necrosis of the myofilaments declined, and the cellular edema was relieved markedly in the I/R + MALAT1 siRNA group. It was shown in the results of immunohistochemistry and Western blotting that MALAT1 siRNA could notably reverse the I/R-induced up-regulation of ß-catenin expression (p<0.05). CONCLUSIONS: MALAT1 knockdown can significantly ameliorate the I/R-induced myocardial injury and improve the cardiac function of the rats, whose mechanism is probably correlated with the inhibition of MALAT1 siRNA on ß-catenin. Therefore, MALAT1 siRNA is expected to become a new target for the treatment of myocardial infarction.

Aldose reductase inhibitor Epalrestat alleviates high glucose-induced cardiomyocyte apoptosis via ROS.

OBJECTIVE: To clarify the role of aldose reductase inhibitor (ARI) in the high glucose-induced cardiomyocyte apoptosis and its mechanism. MATERIALS AND METHODS: In this study, H9c2 cardiomyocytes were employed as objects, high-glucose medium as stimulus, and ARI Epalrestat as a therapeutic drug. The cell apoptosis and activity changes of nitric oxide synthase (NOS), NO, and reactive oxygen species (ROS) were evaluated via Hoechst staining, enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR), and Western blotting. In addition, the mitochondrial membrane potential was measured via fluorescence counting. RESULTS: Epalrestat inhibited the activity of AR to improve high glucose-induced oxidative stress in cardiomyocytes, weaken ROS activity, relieve the inhibition on NO activity, alleviate mitochondrial membrane potential damage, reduce the level of high glucose-induced cardiomyocyte apoptosis, and suppress the expression and activity of Caspase-3, thereby preventing high glucose-induced cardiomyocyte apoptosis. CONCLUSIONS: ARI protects against high glucose-induced cardiomyocyte apoptosis.

First evidence of Mycoplasma haemocanis in China.

Previous studies show heamotropic Mycoplasma infection frequently occurs among splenectomized, immuno-suppressive or co-infected dog populations. However, in our study, the detection of 162 blood samples from dogs found 3 healthy, female dogs infected with Mycoplasma haemocanis in southeastern China. These infected dogs were grown in dog breeding center and had a history of tick infestation. This is the first molecular report of M. haemocanis in dogs from China. The 16S rRNA gene was partially sequenced and a phylogenetic tree constructed. Mycoplasma spp. was 99.9%-100% identical to the corresponding gene sequences of M. haemocanis and M. haemofelis available in GenBank. In this study, Mycoplasma spp. was identified as M. haemocanis because the bacterium was obtained from dogs.

More advanced or aggressive colorectal cancer is associated with a higher incidence of "high-grade intraepithelial neoplasia" on biopsy-based pathological examination.

BACKGROUND: Invasion of submucosa (ISM) is required for the pathological diagnosis of colorectal cancer according to the WHO criteria. A large proportion of colorectal cancers may be underdiagnosed as high-grade intraepithelial neoplasia (HGIN) because ISM is not identified in the preoperative biopsy. The aim of this study was to investigate the clinicopathologic features that are associated with missing the diagnosis of ISM in biopsy specimens of invasive colorectal cancer. METHODS: Three hundred and sixteen patients diagnosed with colorectal cancer between January 2007 and December 2008 with well-preserved preoperative biopsy specimens were enrolled in the study. Three hundred and eleven patients had an isolated lesion, and five had two lesions. Biopsy specimens were reevaluated by two senior pathologists. Clinicopathologic features, biopsy pathology and surgical pathology results of all patients were analyzed by univariate and multivariate analyses. RESULTS: ISM was identified in 216 cases (67.3 %) by biopsy-based pathological examination, and missed in 105 (32.7 %) cases, 72 of which were diagnosed as HGIN. Univariate analysis indicated that in colorectal cancer patients with smaller biopsy specimens (P = 0.042), mucinous or signet-ring cell carcinoma (P = 0.003), higher WHO tumor grade (P = 0.001) and positive lymph nodes (P = 0.011), ISM was more likely to be missed. There was a trend toward an increased diagnosis of ISM with the increase in the number of biopsy specimens (P = 0.105). On multivariate logistic regression analysis, smaller biopsy specimens (OR, 1.810; 95 % CI, 1.081-3.032; P = 0.024) and higher WHO tumor grade (OR, 2.073; 95 % CI, 1.046-4.107; P = 0.037) were the only factors associated with failure to identify ISM. CONCLUSIONS: A large number of invasive colorectal cancers are at risk of being underdiagnosed as HGIN by biopsy-based pathology. The smaller the biopsy size, the less likely it is that the muscularis mucosae is included in the specimen. Also, in the more advanced or aggressive colorectal cancers, ISM is more likely to be missed on biopsy, which may be due to the destruction of the muscularis mucosae by more aggressive cancers.

The effects of a fat- and sugar-enriched diet and chronic stress on nonalcoholic fatty liver disease in male Wistar rats.

PURPOSE: The pathogenesis of nonalcoholic fatty liver disease (NAFLD) is still under debate. The aim of this study was to investigate the effects of a long-term fat- and sugar-enriched diet (FSED) and chronic stress (CS) on NAFLD. METHODS: Male Wistar rats were fed on either a standard diet or a FSED and given CS, a random electric foot shock (2 hr/morning and afternoon per day), or not for 12 weeks. After the experimental period, epididymal adipose tissue weight, sign of visceral obesity (VO), and hepatic index (HI) were measured. At sacrifice blood samples and liver were obtained. Histology of the liver was blindly determined by a pathologist. RESULTS: Histopathologically, moderate to severe steatosis, ballooning hepatocytes, and portal or lobules inflammation were observed in the FSED+CS group. However, mild to moderate steatosis with a few portal inflammation in the FSED group and mild steatosis or not with a few portal inflammation in the CS group were found correspondingly. In addition, more severe blood-fat disorder, high HI, fatty metabolic dysfunction, oxidative stress, high expressions of C-reactive protein mRNA and low expressions of peroxisome proliferator-activated receptor alpha mRNA in the liver were also revealed in the FSED+CS group. But, the degree of VO was not different between the FSED and FSED+CS groups. CONCLUSION: The observations strongly suggest that chronic stress can aggravate fat- and sugar-enriched diet-induced NAFLD from steatosis to steatohepatitis in male Wistar rats, although VO is not changed.

Variation of ER status between primary and metastatic breast cancer and relationship to p53 expression*.

OBJECTIVE: Estrogen-dependent growth of breast cancer can be blocked by anti-estrogens. Estrogen receptor (ER) presence in breast cancer implies responsiveness to endocrine therapy. However, for those patients who ultimately develop resistance to endocrine therapy, the mechanisms remain unclear. The present study attempted to compare the expression status of ER mRNA in a series of primary breast tumors with matched metastases and explored the relation between ER and mutant p53 expression. METHODS: In situ hybridization using a digoxigenin-labeled estrogen receptor cDNA probe was employed to determine the expression of ER mRNA in 52 cases of primary tumors and their matched axillary lymph node metastases. Immunohistochemical staining using a monoclonal antibody against ER was also performed. RESULTS: ER expression was observed in 53.8% (28/52) of primary tumors and 48% (25/52) of metastases, while 57.7% (30/52) of primary tumors and 53.8% (28/52) of metastases showed ER mRNA positivity. There were variations in ER status between in situ hybridization and immunohistochemistry measurements and between primary tumors and metastases. Mutant p53 expression was inversely associated with ER-negative, high-grade tumors. CONCLUSIONS: In situ hybridization may be a more specific and sensitive method for determination of ER status than immunohistochemistry. It is possible that the biologic properties of ER change, and these changes may influence tumor response to endocrine therapy. In view of the ER variation, it was suggested that the ER status of metastatic tumors in addition to primary tumors should be taken into consideration in order to better determine the benefit of clinical endocrine therapy.

A comparison of the patterns of laminin expression in fibroadenoma, fibrocystic diseases, pre-invasive and invasive ductal breast carcinoma.

The basement membrane (BM), of which laminin is a major glycoprotein component, is an important barrier to tumour cells which must be breeched before metastatic spread can occur. We have compared the pattern of laminin expression in a range of benign and malignant breast lesions to better understand the process of tumour progression. A total of 162 cases of breast samples, comprising 18 fibroadenomas, 22 cases of fibrocystic disease, 96 cases of invasive ductal carcinoma and 26 carcinomas with intraductal components, were evaluated for laminin expression by a standard immunoperoxidase method on formalin-fixed, paraffin-embedded histological sections, using a commercial antibody against human laminin. The pattern of laminin expression was charted as follows: Type I, > 70% of BM complete/continuous; Type II, > 70% of BM moderately disrupted; Type III, > 70% of BM completely disrupted. The Type I pattern was observed in all cases of fibroadenoma and fibrocystic diseases, and in 77% of intraductal carcinoma components. Various patterns of BM disruption were observed in invasive ductal carcinoma. Severity of BM disruption correlated with histological grade of the carcinomas (P < 0.001). Small-sized tumours, those without lymphatic invasion and lymph node-negative tumours showed more complete patterns of laminin expression. The current study suggests that tumour cells with high histological grade possess an enhanced capacity to disrupt the basement membrane, an important step in the metastatic process. The detection of BM disruption by immunohistochemical staining for laminin is technically easy and may be usefully applied for the differentiation of in situ and microinvasive carcinoma.

A comparison of the pattern of cathepsin-D expression in fibroadenoma, fibrocystic disease, preinvasive and invasive ductal breast carcinoma.

In the metastatic process, proteolytic enzymes play an important role in mediating the passage of cancer cells through the basement membrane and extracellular matrix. We have compared cathepsin-D (CD) expression in a range of benign and malignant breast lesions so as to investigate its role in breast cancer progression. One hundred and sixty-two breast samples, comprising 18 fibroadenomas, 22 fibrocystic disease, 96 invasive ductal carcinoma and 26 lesions with intraductal carcinoma components, were evaluated for CD expression by the standard avidin-biotin-immunoperoxidase complex method on formalin-fixed, paraffin-embedded histological sections using a commercial antibody against human cathepsin-D. Of the invasive ductal carcinomas, 61.5% showed stromal cell CD positivity, whereas 48.9% expressed CD positivity in neoplastic cells. There was significant correlation between neoplastic cell and stromal CD positivity. The prevalences of CD positivity in both neoplastic and stromal cell components were significantly higher (P < 0.05 and P < 0.01, respectively) in histological grade III tumors compared to grades I and II carcinomas. CD expression by either neoplastic or stromal cells did not show significant correlation with patient age and tumor size. Only 15% of intraductal carcinomas were CD positive and expression was limited to neoplastic cells. Neither epithelial nor stromal cells in fibrocystic lesions and fibroadenomas were CD positive, but a weak to moderate positivity was observed within myoepithelial cells in mammary ducts. These findings provide insights into the mechanism whereby tumors with high histological grade mediate invasion into tissue. The role of stromal cells in tumor progression and the means of their recruitment deserve further study.

Public attitudes towards the autopsy in China.

The autopsy rate in China has fallen within the last few years. The attitudes of the public have become an important factor for its decline. Attitudes towards autopsy were surveyed from 400 subjects residing in the 27 provinces of China. Among the 299 survey forms returned, 284 were completed correctly, the remaining 15 being incomplete. The response rate was 71%. More than half of the responding members considered autopsy beneficial. However their perception of autopsy was quite inaccurate and prejudicial, lacking complete information. The two most frequent reasons for not accepting an autopsy were the lack of awareness towards the practice of autopsy and the wariness of body disfigurement after the autopsy was performed. It was suggested that the most appropriate way to change public perception of autopsy was through education. The primary tool for reaching the public was through the media. In addition, the approach to obtaining consent for autopsy should be improved.

Quantitative comparison of apoptosis to cell proliferation and p53 protein in breast carcinomas.

OBJECTIVE: To clarify the correlation between apoptosis and tumor cell proliferative activity in human breast cancer and to investigate their relevance to p53 protein. STUDY DESIGN: Seventy-one breast carcinomas with histologic grading were analyzed, using counting of mitotic activity index (MAI) and apoptotic index (AI) to examine apoptosis and cellular proliferation, which were then compared with the expression of p53 protein by using a semiquantitative immunohistochemical method. RESULTS: Both the mean MAI and AI were significantly higher in the grade 3 groups (18.30 +/- 2.18 SE, 13.58 +/- 1.94) and 2 (11.32 +/- 1.30, 9.96 +/- 1.84) than in the grade 1 groups (8.24 +/- 1.10, 8.30 +/- 2.20) (P < .001). Also, MAI/AI was significantly highest in the grade 3 group (P < .001). A significant correlation was found between MAI and AI (r = .767, P < .01). Positive expression of p53 protein, indicated by distinct nuclear staining, was found in 35 of 71 carcinomas and was related to neither MAI nor AI (P > .05); there was no significant relation between p53-positive scoring and histologic grading (P > .05). CONCLUSION: Apoptosis in breast cancer seems to correlate with proliferative activity assessed by the mitotic index and supports the hypothesis that apoptosis may play a role in the selection of clonal subpopulations with high growth potential but is not regulated by the p53 system. Further research needs to be conducted to elucidate the relation between apoptosis and tumor progression and the significance of p53 in abnormalities in breast cancer.

Spectroscopic Techniques using Synchrotron Radiation and Free-Electron and Conventional Lasers.

Considerable progress in the investigation of the electronic and vibrational properties of atoms, molecules, materials, surfaces and interfaces has been achieved by combining different photon sources of complementary characteristics. In this paper some experimental results obtained recently at LURE by using two synchronized sources, such as the IR free-electron laser (FEL) CLIO, the VUV storage ring FEL, synchrotron radiation and table lasers, are presented. Using CLIO synchronized with a YAG laser allows the investigation of the vibrational properties of adsorbed species by the non-linear optical technique of visible-IR sum (difference) frequency generation, as shown for the adsorption of hydrogen on platinum in the electrochemical environment. The second result reported here relates to the study of the intersubband stimulated emission in GaAs/GaAlAs quantum wells by pump-probe experiments using the two-colour configuration of CLIO. The combination of a mode-locked Ar(+) laser and synchrotron radiation has been used for investigations in a pump-probe arrangement of the ionization of Xe atoms via the resonant state Xe* 5p(5)5d [3/2](1). The final example is a time-resolved core-level spectroscopy study of photoexcited Si(111) 2 x 1 surfaces by using a combination of the naturally synchronized UV storage ring laser and synchrotron radiation.