[Clinical and genetic studies of a family with hereditary angioedema].

Objective: To diagnose a large family of patients with hereditary angioedema, and to study its inheritance pattern and gene locus. Methods: A retrospective analysis was carried out from August 2021 to February 2022 in a proband (female, 48 years old) and 12 family members who underwent medical history collection and laboratory examinations in the Department of Otorhinolaryngology and Head and Neck Surgery, the Second Hospital of Shanxi Medical University. The clinical data of members and non-affected members [including 7 males and 5 females, aged 12-78 (median 24) years old], were drawn a family map while confirming the diagnosis. Whole exome sequencing technology was used to detect the genetic sequence of the proband and to verify its family members to map the genetic pedigree of the mutation. Results: The inheritance pattern of the family was autosomal dominant, and 8 members of the family were diagnosed with hereditary angioedema by laboratory examination, including 7 cases of type I and 1 case of type Ⅱ. Whole exome sequencing analysis was performed on 2 patients with 2 phenotypes, and it was found that they both carried the same pathogenic mutation locus, which was c.890-2A>G. The family members were verified by next-generation sequencing, and it was found that all members of the family who had a history of edema contained this mutation site, while the younger brother of the proband who had no history of edema did not have this mutation. Conclusion: Both type Ⅰ and type Ⅱ phenotypes are present in this hereditary angioedema family, and the mutation of SERPING1 gene c.890-2A>G causes the onset of each patient in this family.

LINC00641 induces the malignant progression of colorectal carcinoma through the miRNA-424-5p/PLSCR4 feedback loop.

OBJECTIVE: To illustrate the role of LINC00641 in inducing the malignant progression of colorectal cancer (CRC) through the miRNA-424-5p/PLSCR4 feedback loop. PATIENTS AND METHODS: LINC00641 levels in paired CRC and non-tumoral tissues were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Its prognostic potential in CRC was assessed by Kaplan-Meier method. Changes in proliferative and migratory abilities of HCT116 and SW620 cells transfected with si-LINC00641 were evaluated by 5-Ethynyl-2'- deoxyuridine (EdU), cell counting kit-8 (CCK-8) and transwell assay. The feedback loop LINC00641/miRNA-424-5p/PLSCR4 was identified through Dual-Luciferase reporter assay and its involvement in CRC progression was finally explored by rescue experiments. RESULTS: LINC00641 was upregulated in CRC tissues, which was an unfavorable factor to the overall survival of CRC. Proliferative and migratory abilities of HCT116 and SW620 cells were inhibited by knockdown of LINC00641. LINC00641 could competitively bind miRNA-424-5p, thereby abolishing its inhibitory effect on PLSCR4 expression. Knockdown of PLSCR4 could inhibit proliferative and migratory abilities of HCT116 and SW620 cells. CONCLUSIONS: LINC00641 stimulates proliferative and migratory abilities of CRC through the miRNA-424-5p/PLSCR4 feedback loop.

Correlation between microRNA-766 expression in patients with advanced gastric cancer and the efficacy of platinum-containing chemotherapy.

OBJECTIVE: We aimed at observing the correlation between microRNA-766 expression and the efficacy of platinum-containing chemotherapy in patients with stage IV gastric cancer (GCa). PATIENTS AND METHODS: Tissue specimens were obtained from 100 patients with stage IV GCa who received platinum-based chemotherapy, and microRNA-766 expression in these samples was examined by quantitative real-time polymerase chain reaction (qPCR) analysis. Survival analysis was carried out through Kaplan-Meier test. The influencing factors of survival were assessed through COX univariate and multivariate regression. RESULTS: GCa tissues contained significant lower expression of microRNA-766 than adjacent tissues. The degree of tumor differentiation and peritoneal metastasis were confirmed to have great relevance to microRNA-766 level. Patients with high microRNA-766 expression have better chemotherapy efficacy and longer progression-free survival. CONCLUSIONS: Our study shows for the first time that the highly expressed microRNA-766 in tumor tissues of patients with stage Ⅳ GCa predicts better platinum-containing chemotherapy efficacy and prognosis.

Roles of circ-CSPP1 on the proliferation and metastasis of glioma cancer.

OBJECTIVE: The aim of this study was to explore the association between circ-CSPP1 and the proliferation, invasion, and migration of glioma cancer (GC). PATIENTS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was used to detected circ-CSPP1 expression in GC tissues and cells. Subsequently, siRNA was transfected to suppress circ-CSPP1 expression in vitro. Cell counting kit-8 (CCK-8) assay, colony formation assay, and 5-Ethynyl-2'-deoxyuridine (EdU) staining assay were performed to examine the proliferation of GC cells. Meanwhile, transwell assay was conducted to determine the invasion and migration of GC cells. Furthermore, Western blot assay was conducted to analyze the protein expressions of E-cadherin, N-cadherin, and Vimentin. RESULTS: Circ-CSPP1 expression was significantly up-regulated both in GC tissues and cells. GC cells with low expression of circ-CSPP1 showed significantly reduced proliferation, invasion, and migration abilities. In addition, up-regulated E-cadherin protein expression, along with down-regulated N-cadherin and Vimentin protein expressions were observed in GC cells with circ-CSPP1 siRNA treatment. CONCLUSIONS: Circ-CSPP1 promoted the proliferation, invasion, and migration of GC cells.

Molecular mechanisms of lipid metabolism disorder in livers of ewes with pregnancy toxemia.

Pathogenesis of pregnancy toxemia (PT) is believed to be associated with the disruption of lipid metabolism. The present study aimed to explore the underlying mechanisms of lipid metabolism disorder in the livers of ewes with PT. In total, 10 pregnant ewes were fed normally (control group) whereas another 10 were subjected to 70% level feed restriction for 15 days to establish a pathological model of PT. Results showed that, as compared with the controls, the levels of blood ß-hydroxybutyrate (BHBA), non-esterified fatty acids (NEFAs) and cholesterol were greater (P<0.05) and blood glucose level was lower (P<0.05) in PT ewes. The contents of NEFAs, BHBA, cholesterol and triglyceride were higher (P<0.05) and glycerol content was lower (P<0.05) in hepatic tissues of PT ewes than those of the controls. For ewes with PT, excessive fat vacuoles were observed in liver sections stained with hematoxylin-eosin; furthermore, inner structures of hepatocytes including nuclei, mitochondria and endoplasmic reticulum were damaged seriously according to the results of transmission electron microscope. Real-time PCR data showed that compared with the controls, the expression of hepatic genes involved in fatty acid oxidation (FAO) and triglyceride synthesis (TGS) was enhanced (P<0.05) whereas that related to acetyl-CoA metabolism (ACM) was repressed (P<0.05) in PT ewes. Generally, our results showed that negative energy balance altered the expression of genes involved in FAO, ACM and TGS, further caused lipid metabolism disorder in livers, resulting in PT of ewes. Our findings may provide the molecular basis for novel therapeutic strategies against this systemic metabolic disease in sheep.

First Report of Fusarium oxysporum and F. solani Causing Fusarium Dry Rot of Carrot in China.

Carrot (Daucus carota L.) is an economically important vegetable crop in China. In August 2008, a disease was observed on carrot in Inner Mongolia. The symptoms appeared as dry rot lesions on root surface, expressing light brown cankers with defined rounded or irregular shapes (1,3). The average disease incidence was up to 80% in Tuo Ke Tuo County. The disease has been a serious problem in these two counties since then, especially where consecutive carrot cropping was practiced. Carrot roots with typical dry rot symptoms were washed with tap water. Root tissues near the margin of necrotic lesions were excised, surface sterilized with 1% NaOCl for 3 min, and rinsed with sterile distilled water three times. The disinfected tissue was placed on potato dextrose agar (PDA) in a petri dish. Plates were incubated at 25 ± 1°C in the dark for 4 days. Fusarium single spore isolates were obtained from characteristic colonies (1). Three isolates (CF1, CF2, and CF3) were used for further study. The isolates were identified as Fusarium spp. on the basis of microscopic morphology on PDA. CF1 produced pink pigment, abundant falciform macroconidia of 14.7 to 38.2 × 4.5 to 5.7 µm with 2 to 3 septates, and elliptic microconidia of 7.5 to 15.1 × 3.3 to 5.4 µm with none or one septate. CF2 and CF3 produced light blue pigment, abundant falciform macroconidia of 16.4 to 34.4 × 4.0 to 6.1 µm with 2 to 3 septates, and elliptic microconidia of 6.7 to 10.7 × 3.0 to 4.9 µm with none or one septate. They were further identified and confirmed by PCR. The PCR involved amplifying the internal transcribed spacer (ITS) region of ribosomal DNA using genomic DNA as the template with universal primers ITS1 and ITS4 (2). The PCR products were sequenced. BLAST analysis of these sequences against the GenBank database determined the taxonomy of the isolates. The sequence of CF1 was 99% identical to F. oxysporum (Accession No. KC594035); sequences of CF2 and CF3 were 99% identical to F. solani (KC215123). To confirm the pathogenicity of the isolates, mature carrot roots (cv. Hong Ying 2) were inoculated with mycelial plugs (5 mm in diameter) cut from the margin of actively growing colonies on PDA plates. One mycelial plug was placed on each carrot root, with the mycelial side facing the root. PDA plugs were used for controls. Each treatment had five replicates. The inoculated roots were incubated in a humid chamber (90% RH) at 25°C. Four days after incubation, mycelia of the isolates developed and covered most of the surface of carrot roots, and brown rot lesions were observed on all inoculated roots, while the controls remained symptomless. This experiment was repeated. In another trial, carrot seeds (cv. Hong Ying 2) were sown in sterilized soil in pots (30 × 25 cm opening) with 15 seeds per pot. The soil was infested with either CF1, CF2, or CF3 by adding spore suspension to make the final concentration of 1 × 104 CFU/g soil. Plants grown in non-infested soil served as controls. There were three replicates per treatment. All the treated pots were placed in a field. After 13 weeks, the same symptoms of dry rot were observed as previously described. No symptoms were observed on the control plants. The trial was repeated. Symptomatic tissues from the inoculated roots were sampled and the pathogen was re-isolated, and identified using PCR. To our knowledge, this is the first report of F. oxysporum and F. solani causing dry rot of carrot in China. References: (1) H. Abe et al. Annual Report of the Society of Plant Protection of North Japan, 48:106-108, 1997. (2) X. Lu. Plant Dis. 97:991, 2013. (3) A. F. Sherf and A. MacNab. Pages 138-139 in: Vegetable Diseases and Their Control. John Wiley & Sons, Inc., 1986.

Effects of diagnostic contrast-enhanced ultrasound on permeability of placental barrier: a primary study.

OBJECTIVE: To evaluate the effects of contrast-enhanced ultrasound (CEU) on the permeability of placental barrier primarily. METHODS: A total of 60 pregnant Sprague Dawley (SD) rats were divided into 10 groups, including six groups of microbubbles-enhanced ultrasound (varied mechanical index (MI) of 0.13, 1.0 and 1.4 with continuous and intermittent insonation respectively) (US+MB), two groups of ultrasound insonation only (continuous and intermittent insonation respectively) (US), the group of microbubbles only (MB) and the control group. Evans blue (EB), as the tracer, was intravenously injected before treatment. The EB in placenta and fetus was observed under fluorescence microscope and analyzed quantitatively. The EB amount was compared between groups and between placenta and fetus. Lanthanum nitrate-tracing transmission electron microscope examination was performed to observe the distribution of lanthanum in the placenta and fetus. RESULTS: Observed by naked eye, the plancenta was dyed into deep blue while there was no sign of dyeing to the fetus in all groups. Under fluorescence microscope, the red fluorescence radiated by EB was observed in placenta but not in fetus. The EB amount in placenta, insonated by microbubbles-enhanced ultrasound of varied MI, was higher significantly than that in MB, US and control group (all P<0.01) while there was no difference between the latter three groups. And in each group, EB amount was much higher in placenta than that in fetus (P<0.01). The lanthanum particles deposited in the intercellular space in the syntrophoblast while there was no lanthanum presented in the cytotrophoblast in all groups. CONCLUSION: Our findings suggest that diagnostic CEU with SonoVue will not increase the permeability of placenta to the macromolecules larger than albumin, although it may affect placenta.

[Production of oil-processing compounds by microorganisms from the Daqing oil field, China]. / Obrazovanie neftevytesniaiushchikh soedinenii mikroorganizmami iz neftianogo mestorozhdeniia Datsin (KNR).

Twenty pure cultures isolated from formation waters of the Daqing oil field were studied with respect to their capacity to produce surface-active compounds in media with individual hydrocarbons, lower alcohols, and fatty acids. Aerobic saprotrophic bacteria belonging to the genera Bacillus, Brevibacillus, Rhodococcus, Dietzia, Kocuria, Gordonia, Cellulomonas, Clavibacter, Pseudomonas, and Acinetobacter decreased the surface tension of cultivation media from 55-63 to 28-44 mN/m. Strains of Bacillus cereus, Rhodococcus ruber, and Bacillus licheniformis produced biosurfactants most actively. Bacteria of the genera Rhodococcus, Dietzia, Kocuria, and Gordonia produced exopolysaccharides in media with hydrocarbons. Culture liquids of the strains of R. ruber and B. licheniformis exhibited oil-releasing effect. Thus, the Daqing oil field is inhabited by aerobic bacteria capable of producing effective oil-releasing agents.

[The medium conditioned incubation with rat heart cells maintains the properties of ES cells].

17 kinds of conditioned media were selected by plating tests, propagation and other tests with two cell lines, C-19-2 and MESPU-13. The results suggested that the rat heart cells conditioned media (RH-CM) can inhibit the spontaneous differentiation effectively, maintain the normal diploid karyotypes and promote adherence and proliferation of mouse ES cells. The ES cells were propagated in RH-CM to the 20th passage remaining their pluripotent in vivo and in vitro differentiation ability. RH-CM can be used as supplement of ES cell media. ES cells which are cultured in media containing 70% RH-CM and on PMEF feeder can maintain their undifferentiation state and diploid karyotype. RT-PCR detection suggested that there was mLIF expression in rat heart cells.

Porcine inhibin: initial fractionation as a high molecular weight complex.

A high molecular weight complex or aggregate of inhibition was obtained by chromatography of porcine follicular fluid in Fractogel TSK65F. Recovery of activity was good (usually 80-100%), but only 30-60% was recovered as a high molecular weight complex (greater than 160,000) free of albumin and gamma globulin (the two major proteins in follicular fluid). The balance of the activity was distributed in the gamma globulin-albumin region of the chromatogram (i.e., 160,000 down to 65,000 daltons). Distribution in this region of the chromatogram in part reflected the prior processing of the sample (e.g., it was augmented by ethanol or acetone precipitation prior to chromatography). The utility of Fractogel chromatography lies in its ability to resolve a large portion of the inhibin activity from the major proteins (albumin and gamma globulin), plus an efficient recovery of activity. Maximum purification on the Fractogel chromatograms was approximately 20-fold. This product and the other Fractogel fractions were tested for protease activity by a sensitive slab gel procedure. All fractions contained detectable protease activity that could potentially affect inhibin activity during further fractionations. This was shown with a protease fraction isolated from porcine follicular fluid by affinity chromatography. When added to a partially purified inhibin preparation, this protease fraction destroyed 77% of the inhibin activity.