To investigate the function of ALK3 gene, the gene regulation and the signaling pathway related to ventricular septum defect during heart development. The model mice with ALK3 gene knock-out via alpha-MHC-Cre/lox P system were bred. The mRNA expression level of control group was compared with that of experiment group and ALK3 downstream genes were screened using PCR-select cDNA subtraction microarray. The mRNA of control group was extracted from E11.5 normal mouse hearts, and that of experiment group, from E11.5 hearts of mice with alpha-MHC Cre(+/-) ALK3(F/+) genotype. It was found that the mice with ALK3 gene knock-out produced heart defects involving the interventricular septum. The platelet-activating factors acetylhydrolase and the transcription factor Pax-8 and so on, were down-regulated. However, the Protein Tyrosine Kinase (PTK) of Focal Adhesion Kinase (FAK) subfamily and beta subtype protein 14-3-3 were up-regulated in the alpha-MHC Cre(+/-) ALK3(F/-) mice. These data provide support that ALK3 gene played an important role during heart development. The platelet-activating factors acetylhydrolase and Pax-8 genes could be important ALK3 downstream genes in the BMP signaling pathway during interventricular septum development. PTK and beta subtype protein 14-3-3 might be regulatory factors in this pathway.
1-Alkyl-2-acetylglycerophosphocholine Esterase ; genetics ; metabolism ; 14-3-3 Proteins ; genetics ; metabolism ; Animals ; Bone Morphogenetic Protein Receptors, Type I ; genetics ; metabolism ; Genotype ; Heart Septal Defects, Ventricular ; genetics ; Mice ; Mice, Knockout ; Oligonucleotide Array Sequence Analysis ; PAX8 Transcription Factor ; Paired Box Transcription Factors ; genetics ; metabolism ; Protein-Tyrosine Kinases ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; genetics ; physiology

Background Indirect traumatic optic neuropathy (TON) is an acute injury of the optic nerve associated with severe visual dysfunction,which may be a result of secondary mechanical injury and vascular disorder of the optic nerve due to trauma.We analyzed the natural course of axonal loss and blood flow disturbances in patients with indirect TON to find a possible therapeutic window.Methods A cohort of 54 patients with indirect TON recruited between October 2008 and October 2010 at Beijing Tongren Hospital was retrospectively analyzed.The patients were divided into no light perception group (NLP) and better than NLP (btNLP) group.Specifically,the thickness of the retinal nerve fiber layer (RNFL) measured by spectral domain optical coherence tomography (SD-OCT),and hemodynamic parameters of the ophthalmic artery (OA),central retinal artery (CRA) and posterior ciliary artery (PCA) were determined.Results Two weeks after injury,there was a statistically significant decrease in the thickness of RNFL in the btNLP group as compared with the fellow control eyes (P ＜0.05).In contrast,in the NLP group,RNFL thickness slightly increased for 2 weeks following injury,then overtly reduced after 4 weeks (P ＜0.05).Peak systolic velocity (PSV) of CRA was significantly decreased 4 weeks after injury (P ＜0.05) in both the NLP group and btNLP group (P ＜0.05).The thickness of RNFL in the NLP group was negatively correlated with PSV of CRA after 1 week of injury (P ＜0.05,r=-0.962).Conclusions SD-OCT is a useful supplement in detecting the axonal loss in TON.The dynamic change of the thickness of RNFL appears to correlate with the hemodynamic disturbances in the natural course of TON.The first 2 weeks following an injury is critical and should be considered as the therapeutic window for TON patients.

OBJECTIVETo evaluate the clinical value of double contrast-enhanced ultrasonography using oral and intravenous contrast agents in preoperative staging of gastric cancer.

METHODSSixty-two patients with biopsy-proven gastric cancer were enrolled into this study, and were examined by double contrast-enhanced gastric ultrasonography preoperatively. The results were compared with postoperative pathologic findings.

RESULTSThe accuracy of oral contrast-enhanced gastric ultrasonography and double contrast-enhanced ultrasonography in determining the T stage of gastric cancer was 72.9% (T1: 66.7%, T2: 60.0%, T3: 76.9%, T4: 71.4%) and 88.1% (T1: 66.7%, T2: 80.0%, T3: 89.7%, T4: 100%), respectively, with a statistically significant difference between the two methods (P = 0.036). The sensitivity, specificity, accuracy and Youden index of oral contrast-enhanced gastric ultrasonography and double contrast-enhanced ultrasonography in assessment of lymph node metastasis were 74.5%, 66.7%, 72.9%, and 0.41 versus 89.4%, 75.0%, 86.4%, 0.76, respectively. No significant difference in the accuracy of assessment for lymph node metastasis was observed (P > 0.05).

CONCLUSIONDouble contrast-enhanced ultrasonography is useful for preoperative staging of gastric cancer, especially for T staging.

Adult ; Aged ; Contrast Media ; Endosonography ; methods ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; methods ; Preoperative Care ; Prospective Studies ; Stomach Neoplasms ; diagnostic imaging ; pathology ; surgery ; Sulfur Hexafluoride

OBJECTIVEConventional deletion of ALK3, also termed as bone morphogenetic protein (BMP) receptor IA, in mice might result in early embryonic lethality. To investigate the function of ALK3 in cardiac development, the cardiac-specific deletion of ALK3 in mice was made by Dr. Schneider, using Cre recombinase driven by the alpha-MHC promoter that Dr. Fukushipe worked out. Such specific deletion of ALK3 caused death in mid-gestation with defects in the trabeculae, interventricular septum, and endocardial cushion. Since ALK3 is not a cardiac-specific gene, it is extremely important to identify ALK3 downstream genes.

METHODSAlpha-MHC Cre+/-, ALK3 F/- and alpha-MHC Cre+/-, ALK3 F/+ embryos were obtained after 20 alpha-MHC Cre+/-, ALK3 +/- mice and 20 ALK3 F/F mice were mating. The ALK3 downstream genes were screened using microarray made in Germany that could identify 25000 genes in mouse. Two populations of mRNA, one derived from the embryonic heart (11.5 days) of alpha-MHC Cre+/-, ALK3 F/- mice, and the other derived from the alpha-MHC Cre+/-, ALK3 F/+ mice, were compared. Cardiac-specific ALK3 downstream genes were identified using real time quantitative RT-PCR and in situ hybridization.

RESULTSThe expression of 12 genes, such as Pax-8 and Hox-3.5 were down-regulated in alpha-MHC Cre+/-, ALK3 F/- mouse heart. The expression of 16 genes including Ras-related protein Rab-5b and EPS-8 protein was up-regulated in the group of alpha-MHC Cre+/-, ALK3 F/-. It was found that the Box protein Pax-8 gene was down-regulated by 7.1 fold (P < 0.001) in the alpha-MHC Cre+/-, ALK3 F/- mice by real time quantitative RT-PCR. It was also revealed that the Box protein Pax-8 gene was expressed stronger in alpha-MHC Cre+/-, ALK3 F/+ than alpha-MHC Cre+/-, ALK3 F/- E11.5 days mouse heart by means of in situ hybridization.

CONCLUSIONThe Box protein Pax-8 gene is an important and cardiac-specific ALK3 downstream gene in the BMP signaling pathway during inter-ventricular septum development.

Animals ; Bone Morphogenetic Protein Receptors ; DNA-Binding Proteins ; genetics ; Down-Regulation ; Female ; Heart ; embryology ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Myocardium ; metabolism ; pathology ; Nuclear Proteins ; Oligonucleotide Array Sequence Analysis ; PAX8 Transcription Factor ; Paired Box Transcription Factors ; Receptors, Growth Factor ; genetics ; physiology ; Reverse Transcriptase Polymerase Chain Reaction ; Trans-Activators ; genetics

OBJECTIVETo investigate the diagnostic value of contrast-enhanced ultrasonography (CEUS) in preoperative T-staging of gastric cancer.

METHODSA sulfur hexafluoride-filled microbubble ultrasound contrast agent and a continuous real-time imaging technique of contrast pulse sequencing were used. Normal gastric wall was examined by CEUS in 8 healthy volunteers and the results were compared with the findings on multislice computed tomography. Sixty-two patients with gastric cancer proved by biopsies who received preoperative CEUS examination were involved in this study, and the CEUS result was compared with postoperative pathological findings.

RESULTSThe normal gastric wall presented a one-layer structure in the portal venous phase and a three-layer structure in the arterial and equilibrium phase including a slightly hyper-enhanced inner layer, a hypo-enhanced intermediate layer, and a markedly hyper-enhanced outer layer, which corresponded histologically to the mucosal, submucosal, and muscular-serosal layer, respectively. The accuracy of transabdominal ultrasonography and CEUS in determining the T stage of gastric cancer was 72.9% and 88.1% respectively, and the difference was statistically significant (chi(2)=4.37, P=0.036).

CONCLUSIONSCEUS shows the normal gastric wall as a one- or a three-layer structure, which provides a theory base for CEUS in preoperative T-staging of gastric cancer. CEUS is a useful diagnostic method for preoperative T-staging of gastric cancer.

Adult ; Aged ; Contrast Media ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; methods ; Stomach Neoplasms ; diagnostic imaging ; pathology ; Sulfur Hexafluoride ; Ultrasonography ; methods

BACKGROUNDIndirect traumatic optic neuropathy (TON) is an acute injury of the optic nerve associated with severe visual dysfunction, which may be a result of secondary mechanical injury and vascular disorder of the optic nerve due to trauma. We analyzed the natural course of axonal loss and blood flow disturbances in patients with indirect TON to find a possible therapeutic window.

METHODSA cohort of 54 patients with indirect TON recruited between October 2008 and October 2010 at Beijing Tongren Hospital was retrospectively analyzed. The patients were divided into no light perception group (NLP) and better than NLP (btNLP) group. Specifically, the thickness of the retinal nerve fiber layer (RNFL) measured by spectral domain optical coherence tomography (SD-OCT), and hemodynamic parameters of the ophthalmic artery (OA), central retinal artery (CRA) and posterior ciliary artery (PCA) were determined.

RESULTSTwo weeks after injury, there was a statistically significant decrease in the thickness of RNFL in the btNLP group as compared with the fellow control eyes (P < 0.05). In contrast, in the NLP group, RNFL thickness slightly increased for 2 weeks following injury, then overtly reduced after 4 weeks (P < 0.05). Peak systolic velocity (PSV) of CRA was significantly decreased 4 weeks after injury (P < 0.05) in both the NLP group and btNLP group (P < 0.05). The thickness of RNFL in the NLP group was negatively correlated with PSV of CRA after 1 week of injury (P < 0.05, r = -0.962).

CONCLUSIONSSD-OCT is a useful supplement in detecting the axonal loss in TON. The dynamic change of the thickness of RNFL appears to correlate with the hemodynamic disturbances in the natural course of TON. The first 2 weeks following an injury is critical and should be considered as the therapeutic window for TON patients.

Adult ; Female ; Humans ; Male ; Middle Aged ; Nerve Fibers ; physiology ; Optic Nerve ; physiology ; Optic Nerve Injuries ; physiopathology ; Retinal Neurons ; physiology ; Retrospective Studies ; Tomography, Optical Coherence

OBJECTIVES: To explore the optimal maintenance dose of caffeine citrate for preterm infants requiring assisted ventilation and caffeine citrate treatment. METHODS: A retrospective analysis was performed on the medical data of 566 preterm infants (gestational age ≤34 weeks) who were treated and required assisted ventilation and caffeine citrate treatment in the neonatal intensive care unit of 30 tertiary hospitals in Jiangsu Province of China between January 1 and December 31, 2019. The 405 preterm infants receiving high-dose (10 mg/kg per day) caffeine citrate after a loading dose of 20 mg/kg within 24 hours after birth were enrolled as the high-dose group. The 161 preterm infants receiving low-dose (5 mg/kg per day) caffeine citrate were enrolled as the low-dose group. RESULTS: Compared with the low-dose group, the high-dose group had significant reductions in the need for high-concentration oxygen during assisted ventilation (P=0.044), the duration of oxygen inhalation after weaning from noninvasive ventilation (P<0.01), total oxygen inhalation time during hospitalization (P<0.01), the proportion of preterm infants requiring noninvasive ventilation again (P<0.01), the rate of use of pulmonary surfactant and budesonide (P<0.05), and the incidence rates of apnea and bronchopulmonary dysplasia (P<0.01), but the high-dose group had a significantly increased incidence rate of feeding intolerance (P=0.032). There were no significant differences between the two groups in the body weight change, the incidence rates of retinopathy of prematurity, intraventricular hemorrhage or necrotizing enterocolitis, the mortality rate, and the duration of caffeine use (P>0.05). CONCLUSIONS This pilot multicenter study shows that the high maintenance dose (10 mg/kg per day) is generally beneficial to preterm infants in China and does not increase the incidence rate of common adverse reactions. For the risk of feeding intolerance, further research is needed to eliminate the interference of confounding factors as far as possible.
Caffeine/therapeutic use* ; Citrates ; Humans ; Infant ; Infant, Newborn ; Infant, Premature ; Respiration, Artificial ; Retrospective Studies

Objective To explore the sensitivity of Culex pipiens pallens to common chemical insecticides in the southwestern region of Shandong Province, so as to provide a theoretical basis for the development of reasonable and effective mosquito control measures. Methods The resistance of Cx. pipiens pallens larvae to 5 chemical insecticides, such as cypermethrin, deltamethrin, DDVP, propoxur, and acetofenate were tested by using the WHO biological test method in 2018, and the co-toxicity coefficients after compounding the above-mentioned insecticides were tested by using a drug compounding method. Results The resistance indexes of Cx. pipiens pallens to cypermethrin, deltamethrin, DDVP, propoxur, and acetofenate in 3 cities were 144.43–557.54, 118.17–445.33, 6.44–19.00, 2.37–8.10, and 0.88–2.98, respectively, and expect the difference between the DDVP resistances of Cx. pipiens pallens in Jining City and Heze City was not statistically significant (P > 0.05), all the other differences were statistically significant (all P < 0.05). The synergistic coefficients of cypermethrin + DDVP, cypermethrin + propoxur, DDVP + acetofenate, and propoxur + acetofenate were 199.58 – 456.95, 190.56 – 292.37, 123.32 – 319.24, and 192.31 – 367.32, respectively. The lower synergism was observed by using the mixture of DDVP + propoxur (synergistic coefficient: 99.87–108.36) . Conclusions After decades of chemical control, Cx. pipiens pallens in the southwestern region of Shandong Province has produced different degrees of resistance to common chemical insecticides. Therefore, comprehensive control measures should be taken to control mosquito breeding and prevent the development of insecticide resistance.

Objective:To compare the differences in the prevalence of mild micro-hepatic encephalopathy (MHE) among patients with cirrhosis by using the psychometric hepatic encephalopathy score (PHES) and the Stroop smartphone application (Encephal App) test.Methods:This prospective, multi-center, real-world study was initiated by the National Clinical Medical Research Center for Infectious Diseases and the Portal Hypertension Alliance and registered with International ClinicalTrials.gov (NCT05140837). 354 cases of cirrhosis were enrolled in 19 hospitals across the country. PHES (including digital connection tests A and B, digital symbol tests, trajectory drawing tests, and serial management tests) and the Stroop test were conducted in all of them. PHES was differentiated using standard diagnostic criteria established by the two studies in China and South Korea. The Stroop test was evaluated based on the criteria of the research and development team. The impact of different diagnostic standards or methods on the incidence of MHE in patients with cirrhosis was analyzed. Data between groups were differentiated using the t-test, Mann-Whitney U test, and χ2 test. A kappa test was used to compare the consistency between groups. Results:After PHES, the prevalence of MHE among 354 cases of cirrhosis was 78.53% and 15.25%, respectively, based on Chinese research standards and Korean research normal value standards. However, the prevalence of MHE was 56.78% based on the Stroop test, and the differences in pairwise comparisons among the three groups were statistically significant (kappa = -0.064, P < 0.001). Stratified analysis revealed that the MHE prevalence in three groups of patients with Child-Pugh classes A, B, and C was 74.14%, 83.33%, and 88.24%, respectively, according to the normal value standards of Chinese researchers, while the MHE prevalence rates in three groups of patients with Child-Pugh classes A, B, and C were 8.29%, 23.53%, and 38.24%, respectively, according to the normal value standards of Korean researchers. Furthermore, the prevalence rates of MHE in the three groups of patients with Child-Pugh grades A, B, and C were 52.68%, 58.82%, and 73.53%, respectively, according to the Stroop test standard. However, among the results of each diagnostic standard, the prevalence of MHE showed an increasing trend with an increasing Child-Pugh grade. Further comparison demonstrated that the scores obtained by the number connection test A and the number symbol test were consistent according to the normal value standards of the two studies in China and South Korea ( Z = -0.982, -1.702; P = 0.326, 0.089), while the other three sub-tests had significant differences ( P < 0.001). Conclusion:The prevalence rate of MHE in the cirrhotic population is high, but the prevalence of MHE obtained by using different diagnostic criteria or methods varies greatly. Therefore, in line with the current changes in demographics and disease spectrum, it is necessary to enroll a larger sample size of a healthy population as a control. Moreover, the establishment of more reliable diagnostic scoring criteria will serve as a basis for obtaining accurate MHE incidence and formulating diagnosis and treatment strategies in cirrhotic populations.