Objective To explore the value of coronary flow reserve(CFR) evaluating myocardial ischemia measured by adenosine triphosphate (ATP) stress transthoracic Doppler echocardiography (TTDE),and the feasibility of CFR to predict coronary stenosis.Methods Fifty-four patients suffering chest pain with known or suspected coronary artery disease were performed ATP stress TTDE to measure resting and maximum expansion coronary blood flow velocity and calculate CFR.all patients were performed by coronary angiography (CAG) and single-photon emission computed tomography (SPECT) myocardial perfusion imaging.Results ① To evaluate myocardial ischemia,there was not statistical significant difference between non-invasive CFR and SPECT myocardial perfusion imaging(P ＞0.05).CFR≤2.0 was the best cutoff value for evaluating myocardial ischemia which yielded a sensitivity of 93.3 ％ and specificity of 89.7％.②Coronary artery stenosis was negatively correlated with CFR (P ＜0.001).ROC curve analysis demonstrated that CFR≤ 1.60 yielded a sensitivity of 92.3％ and specificity of 73.3％ to predict coronary stenosis significantly.Conclusions CFR measured by ATP stress TTDE can evaluate myocardial ischemia of coronary artery disease and predict LAD significant stenosis before CAG.Using CFR and CAG has important clinical value for choosing treatment of stable coronary artery disease.

OBJECTIVE:To investigate the effect of Danggui shaoyao powder (DSS) on uterine structure and expressions of estrogen receptorα(ERα),estrogen receptorβ(ERβ)in uterine cavity epithelium and matrix in model rats in perimenopausal peri-od. METHODS:40 female SD rats were randomly divided into sham operation group (normal saline),model group (normal sa-line),DSS low-dose,medium-dose,high-dose groups(1.94,3.87,7.44 g/kg),8 in each group. Except that rats in sham opera-tion group received resection of fat nearby ovaries,rats in other groups received resection of bilateral ovaries to induce models in perimenopausal period. After modeling,rats were intragastrically administrated once a day,for 8 weeks. After administration,wet mass of uterine was weighted. Changes in uterine morphology and structure of rats were observed,expressions levels of ERα and ERβ in uterine cavity epithelium and matrix were determined. RESULTS:Compared with sham operation group,rats in model group showed low columnar in endometrial columnar epithelium,lamina propria layer,muscular layer and serous layer were signifi-cantly atrophied,stromal cells had obvious nuclear condensation. There was marked decrease in uterine wet mass,uterine cavity ar-ea and endometrial thickness as well as the number of uterine glands(P<0.01),and the expression levels of ERαand ERβin uter-ine cavity epithelium and matrix were significantly reduced(P<0.01). Compared with model group,the atrophy degree of endome-trium and lamina propria layer had no significant differences in DSS each dose groups. However,lamina propria layer was rich in glands,and there were significant differences in uterine wet mass,uterine cavity area,endometrial thickness,ERα expression level in uterine cavity epithelium and matrix (P>0.05). However, the number of uterine glands in DSS medium-dose,high-dose groups was significantly increased(P<0.01),and ERβexpres-sion level in uterine cavity epithelium and matrix in DSS high-dose group was significantly increased (P<0.05 or P<0.01). CONCLUSIONS:DSS has not obvious effect on im-proving the symptoms of uterine gland atrophy of model rats in perimenopausal period,but it can increase the number ofuterine glands,and the mechanism may be associated with improving the ERβ expression level in uterine cavity epithelium and ma-trix.

Objective To study the effect of CDCA5 silencing on the growth of human breast cancer cell line MDA-MB-435s in vitro and its mechanism.Methods The expression of CDCA5 in breast cancer tissue,the adjacent normal tissue and human breast cancer cell lines MCF-7,MDA-MB-231 and MDA-MB-435s was tested by Western blot.The target sequences of CDCA5 small interfering RNA (siRNA),control siRNA (siRNA-NC) and blank groups were designed.PCR was used to observe the gene silencing effect on CDCA5.The cell viability was measured by MTT assay in siRNA,siRNA-NC and Blank groups.Cell cycle was determined by flow cytometry.Plkl,SA2 and pSA2 protein expression was tested by Western blot.Rssults The expression of CDCA5 was higher in breast cancer tissue than in the adjacent normal tissue.The expression of CDCA5 was higher in human breast cancer cell line MDA-MB-435s than in MCF-7 or MDA-MB-231 cell line.The expression level of CDCA5 was downregulated significantly in siRNA group than other groups.The cell viability was decreased and the cell percentage of G2/M phase was elevated in siRNA group compared with those in siRNA-NC and Blank groups.Inhibition of the CDCA5 gene can suppress the expression of Plk1 and pSA2.Conclusion Silencing CDCA5 expression can effectively inhibit the proliferation of breast cancer cells,whose mechanism may lie in arresting cell cycle in G2/M phase by suppressing CDCA5/Plkl/SA2 signaling pathway.

Objective To investigate the value of CT taking WO3-x@BSA nanoparticles as contrast agents.Methods CT values of WO3-x@BSA nanoparticles and the iohexol contrast agents at different concentrations were measured.When the tube voltage was the same,CT values of the tungsten at different concentrations were measured,and the relationship curve of CT values and the concentrations,the tube voltages and CT values were plotted.The animal models were performed to observe CT imaging in vivo and in vitro.WO3-x@BSA nanoparticles were injected through tail vein and intraperitoneally of rats in vivo.In fresh pig liver,WO3-x@BSA nanoparticles were injected by portal vein.Results CT value of the tungsten was higher than the iodine at the same concentration,and the imaging signal of the tungsten was more significant than the iodine.The amount of WO3-x@BSA was less than the iohexol contrast agent reaching to the same CT value.WO3-x@BSA nanoparticles could reach the organs of various tissues in SD rats through different ways and showing the high density sig nal.WO3-x@BSA nanoparticles were also injected into the fresh in-vitro liver of pig,and could diffuse and distribute along with the blood vessels with good CT imaging.Conclusion As a novel CT contrast agent,WO3-x@BSA nanoparticles can be used to get good CT imaging.

Objective To investigate the expression of BRCA1 in familial thyroid cancer (FTC) and the relationship between BRCA1 expression and clinicopathologic features.Methods The expression of BRCA1 protein and mRNA was detected respectively by immunohistochemistry and RT-PCR in 37 FTC tissues,35 sporadic thyroid carcinoma tissues and 35 normal thyroid tissues.Results BRCA1 mRNA expression was significantly downregulated in FTC (0.210±0.025) compared with sporadic thyroid carcinoma(0.943±0.021) and normal thyroid tissues(1.001±0.087)(P＜0.01).The positive rate of BRCA1 protein expression was significantly lower in FTC(56.8％,21/37) than that in sporadic thyroid carcinoma(85.7％,30/35) and normal thyroid tissues(94.5％,33/35) (P＜0.05).There was a correlation between BRCA1 protein expression and FTC clinicopathologic features (P＜0.05).FTC patients with lower BRCA1 protein expression tended to have higher risk for bilateral carcinoma,lymph node metastasis,capsular invasion,morbidity under 30 years old and more than 2 patients in a family.Conclusions The expression of BRCA1 protein and mRNA is downregnlated in FTC.BRCA1 protein might serve as a potential biomarker for predicating biological behavior and prognosis of FTC.

Objective To investigate the frequencies of the impaired glucose tolerance and diabetes mellitus in a large cohort of active acromegaly and to identify risk factors associated with the presence of diabetes at diagnosis in these patients.Methods This study included 88 patients with newly diagnosed acromegaly.Patients were further divided into normal glucose tolerance (NGT),impaired glucose regulation (IGR),and diabetes (DM) groups according to oral glucose tolerance test or previous history.Insulin sensitivity and β cell function were also evaluated by homeostasis model assessment (HOMA).Logistic regression analysis was used to explore the independent risk factors for diabetes in patients with acromegaly.Results Impaired glucose regulation was found in 25 (28.4％),and DM in 37(42.0％) acromegaly patients.Compared to NGT and IGR patients,higher proportion of DM patients had family history of diabetes (P＜0.05).Compared to NGT group,higher post-OGTT growth hormone (GH) levels were detected in IGR and DM groups.Insulin-like growth factor 1 (IGF-1) levels were higher in IGR group while lower in DM group (P＜0.01).Homeostasis model assessment for β cell function (HOMA-β) was slightly higher in IGR group,but significantly lower in DM group (P ＜ 0.01).Homeostasis model assessment for insulin resistance (HOMA-IR) was slightly higher in IGR and DM groups but without significant difference among 3 groups.In multivariate analyses,family history of diabetes(OR=12.710,95％ CI 1.176-137.30,P=0.036),age(OR=1.106,95％ CI 1.018-1.202,P=0.017),and GH levels(OR=1.075,95％ CI 1.007-1.147,P=0.030) were independent risk factors of diabetes in acromegaly patients.Conclusion Impaired glucose metabolism is present in nearly 70％ of patients at diagnosis of acromegaly,and is associated with age,family history of diabetes,and higher GH levels,but not with IGF-1 levels.

OBJECTIVE: To analyze the clinical phenotype and pathogenic variant in a child diagnosed with mental retardation and microcephaly with pontine and cerebellar hypoplasia (MICPCH). METHODS: Clinical phenotype of the child was reviewed. Whole exome sequencing was carried out for the child. Candidate variant was verified by Sanger sequencing of the family member. RESULTS: The proband manifested dyskinesia, development delay, cerebellar hypoplasia and bilateral hearing impairment. WES results revealed that the proband has carried a pathogenic c.1641_1644delACAA (p.Thr548Trpfs*69) variant of the CASK gene, which was verified by Sanger sequencing to be a de novo variant. CONCLUSION The c.1641_1644delACAA (p.Thr548Trpfs*69) variant of the CASK gene probably underlay the MICPCH in the proband. Above finding has provided a basis for genetic counseling. WES should be considered for the diagnosis of neurological dysplasia.
Cerebellum/abnormalities* ; Child ; Developmental Disabilities ; Family ; Humans ; Mental Retardation, X-Linked ; Microcephaly/genetics* ; Nervous System Malformations

Objective:To assess the renal-protective effect of Elabela (Ela) on diabetic kidney disease (DKD), and explore its potential mechanism.Methods:db/db mice were randomly divided into diabetic group and Ela intervention group, while db/m mice were taken as normal control group. The mice in the Ela intervention group were intraperitoneally injected with Ela-21 5 mg·kg -1·d -1 for 8 weeks. At the end of the experiment, urinary albumin/creatinine ratio (UACR) was measured. The renal pathological changes were observed by HE and PAS staining. The expression of aquaporin 2(AQP2) examined by immunohistochemistry. The level of collage Ⅳ(Col-Ⅳ) and AQP2 in renal tissue was analyzed by Western blot. The human renal tubular epithelial cells (HK-2) were incubated with high glucose medium and further interfered with apelin receptors (APJ)-siRNA. Western blot analysis was used to detect the effect of Ela intervention on Col-Ⅳ and AQP2 expression. Finally, to clarify the possible mechanism of Ela regulating AQP2, the interaction between Ela-induced APJ activation and arginine vasopressin (AVP)-evoked arginine vasopressin receptor 2 (AVPR2) activation was investigated by NanoBit ? technology. Results:(1) Without affecting blood glucose and body weight, Ela intervention significantly reduced the UACR in db/db mice, and attenuate pathological changes of the kidney, as well as expression of Col-Ⅳ and AQP2. (2) Ela treatment could remarkably inhibit the high glucose-induced the expression of Col-Ⅳ and AQP2, which was reversed by interfering with APJ. (3) AVP-induced downstream β-Arrestin-2 signaling transduction via AVPR2 was obviously antagonized by interaction of Ela and APJ, further suggesting that the inhibitory effect of Ela on AQP2 may be related to antagonizing AVP/AVPR2 signaling.Conclusion:Ela exerts renal protection by inhibiting the expression of AQP2 through APJ.

Objective:To explore the effect of endoplasmic reticulum stressed (ER) on colorectal cancer (CRC) cell proliferation and invasion via ATG5-mediated autophagy pathway and the underlying mechanism.Methods:We performed bioinformatics analysis to identify the expression level of PERK, ATF6 and ATG5 in CRC tissues and adjacent tissues and the correlation between PERK and ATG5 expression in CRC tissues.The expression level of PERK in CRC cell lines was examined by qRT-PCR assay. Cell proliferation was quantified by CCK-8.The invasion of the cells was detected by Transwell.Western blot assay was performed to verify the levels of protein. The levels of autophagy were examined by electron microscopy.Results:PERK and ATF6 expression in CRC tissues was higher than that in the adjacent tissues and PERK expression was higher in CRC cells than intestinal mucosal cells. Expression level of PERK in CRC cell lines HCT116,SW480,HT29,LoVo and colonic mucosa cell lines FHC was 1.51±0.04,3.12±0.05,2.19±0.04,2.38±0.06 and 0.98±0.04 ( P<0.001) .The increased expression of PERK promoted CRC cell proliferation and invasion. PERK expression levels was positively associated with ATG5 expression levels ( r=0.52, P<0.001) and overexpression of PERK accelerated the protein expression of ATG5 (1.00±0.04,3.53±0.07, t=74.61, P<0.001) . ATG5 was highly expressed in CRC tissues. Overexpression of ATG5 could promote proliferation,invasion and accelerate autophagy of CRC cells (the number of autophagosomes in the blank control group,the negative control group and ATG5-Overexpression group was 4.33±1.53, 4.00±1.00, 9.67±2.52, and t=3.14,3.62, P=0.035,0.022, respectively) .ATG5 promoted colorectal cancer cell proliferation and invasion through autophagy pathway. Conclusion:ER stressed-CRC cells could promote CRC cell proliferation and invasion through ATG5-mediated autophagy pathway.

Objective:To investigate the clinical significance and possible mechanisms of elevated homocysteine(Hcy) levels in peripheral blood of children with sepsis.Methods:The clinical data of 51 children with sepsis (sepsis group) admitted to PICU at Xuzhou Children′s Hospital from January 2019 to December 2019 were analyzed, and the levels of Hcy in plasma were compared with 50 non-septic children (common infection group) and 50 healthy children (healthy control group) during the same period.The possible mechanism of metabolic disorders about Hcy was analyzed by detecting the levels of the key rate-limiting enzymes cystathionine-β-synthase(CBS) and cystathionine-γ-lyase(CSE), which were in the downstream of metabolism in septic mouse model induced by lipopolysaccharide.Results:The level of Hcy in plasma was (12.62±5.46)μmol/L in sepsis group, which was significantly higher than those in common infection group[(9.42±2.28) μmol/L] and healthy control group[(8.14±1.60) μmol/L]( P<0.05). The level of Hcy in plasma of 12 children with acute kidney injury in sepsis group was significantly higher than that of 39 children without acute kidney injury in sepsis group[(16.48±5.87)μmol/L vs.(11.62±4.74) μmol/L, P<0.05]. The level of Hcy in plasma of six children with acute liver failure in sepsis group was significant higher than that of 45 children without acute liver failure in sepsis group[(18.35±7.10) μmol/L vs.(11.84±4.78) μmol/L, P<0.05]. The level of Hcy in serum significantly increased in septic mouse models ( P<0.01). The transcription and protein expression levels of key rate-limiting Hcy transcription enzymes CBS and CSE in liver and kidney tissues of septic mouse were significantly down-regulated ( P<0.05). Conclusion:The level of Hcy in peripheral blood of children with sepsis increases, which is more obviously in children with acute kidney injury or acute liver injury.When patients developed sepsis, the expression of CBS and CSE will be restrained, leading to disorders related to transsulfuration metabolism and elevated level of Hcy in peripheral blood.