Objective:To improve the primary culture method of rat pulmonary microvascular endothelial cells(PMVECs) and obtain purified PMVECs.Methods:The modified tissue block pasted culture method was used to isolate and culture Wistar rat PMVECs. The morphous of cultured cells were observed by microscopy. The cultured cells were identified by detecting factor Ⅷ related antigen and binding isolectin B4. Results and Conclusion:The morphous of cultured primary PMVECs in vitro showed short fusiform shape or polygon, and the monolayer of cultured cells displayed the shape of pavingstone. But the morphous changed followed the transfer of culture and the change of culture condition. The cultured cells had characterization of binding isolectin B4 and negative immunocytochemical staining for factor Ⅷ related antigen. The cultured PMVECs have good growth state and purity,and can be subcultringed stably.The observation of cell morphous integrating with immunocytochemical staining is a reasonable identification method for PMVECs.

Objective To establish a method of isolation, purification, primary culture and identification of alveolar type Ⅱ epithelial cells(AT-Ⅱ).Methods The AT-Ⅱs were isolated from Wistar rats by trypsin,purified by differential centrifugation, erythrocyte spallation, differential adherence and immune adherence, and identified by observing the morphology of cultured cells under the inverted phase and tannic acid staining. Results and Conclusion The cultured primary AT-Ⅱs in vitro presented single or island form growth, and their shapes were round or elliptical. A great deal of fine particles showed sharp contrast, and were observed in intracytoplasm. The cell nuclei were clear. They were positive for tannic acid staining.The primary culture AT-Ⅱs obtained from improved isolation and purification have good growth state and purity, and are suitable for research in vitro.

Objective To assess the neurophysiological effects of transcranial direct current stimulation (tDCS) of the motor cortex when swallowing.Methods Twenty healthy volunteers had anodal tDCS (a-tDCS), cathodal tDCS (c-tDCS) or sham tDCS applied over the hemisphere with stronger suprahyoid projections, effortful swallowing was performed simultaneously.Suprahyoid motor-evoked potentials (MEPs) on both the stimulated and non-stimulated contra-lateral hemisphere were examined immediately before stimulation and 5,30, 60 and 90 minutes later.The MEPs were normalized and analyzed using two-way repeated measures analysis of variance.Results The tDCS had long-lasting effects on the suprahyoid MEPs bilaterally.There were significant changes in the effect over time.Sham tDCS showed no significant effect.Compared with sham tDCS, a-tDCS significantly increased the excitability of the stimulated hemisphere, but not the non-stimulated projection.Compared with sham tDCS, c-tDCS induced decreased cortical excitability in the stimulated hemisphere but an increase in the non-stimulated projection.Conclusions tDCS during swallowing can alter bilateral swallowing activity in the motor cortex in a polarity-dependent and site-dependent way.A-tDCS enhances the excitability of the stimulated hemisphere while c-tDCS inhibits it ipsilaterally but increases it contralaterally.

Child ; Child, Preschool ; Female ; Follow-Up Studies ; Humans ; Infant ; Male ; Prognosis ; Retrospective Studies ; Treatment Outcome ; Vesico-Ureteral Reflux ; diagnosis ; therapy

OBJECTIVEThe Jiangu decoction is used in the treatment of steroid-induced femoral head necrosis in clinical experiences, which has functions of tonifying kidney and activating blood, and invigorating spleen to remove phlegm. The decoction is mainly composed of Radix Polygoni Multiflori, Rhizoma alismatis Rhizoma Drynariae, haw, medlar, Radix Astragali, radix rehmanniae, angelica, Radix Codonopsis, radix salviae miltiorrhizae, Fructus Ligustri Lucidi, licorice, pharmaceutical composition. This study was designed to investigate the influence of Jiangu decoction on peroxisome proliferator-activated receptor gamma (PPARgamma) in the femoral head of rabbits with steroid-induced femoral head necrosis.

METHODSEighteen adult SPF healthy New Zealand rabbits were divided into 3 groups: control group, model group, Jiangu decoction group. The rabbits of Jiangu decoction group orally received Jiangu decoction suspension with a dose of 10 ml/kg each day and the drug content was 0.719 g/ml. The rabbits in control and model groups were given saline with a dose of 10 ml/kg. The methylprednisolone sodium succinate was injected intramuscularly into left leg with a dose of 40 mg/kg. Then the rabbits were fed continuously for 3 weeks. The glucocorticoid levels, PPARgamma and plasma glucocorticoid levels in the femoral head were measured before and after modeling.

RESULTSBefore model established, the plasma glucocorticoid levels had no significant difference among three groups (P=0.301). At 3 weeks after model established,the plasma glucocorticoid level of rabbits in model group increased compared to the control group (P=0.001); and the plasma glucocorticoid level of rabbits in Jiangu decoction group decreased compared with model group (P=0.001). The glucocorticoid level in the local femoral head of rabbits in model group increased compared to the control group (P=0.001); and the glucocorticoid level in the local femoral head of rabbits in Jiangu decoction group decreased compared with model group (P=0.001). The PPARgamma level in the local femoral head of rabbits in model group increased compared to the control group (P=0.018);and the PPARgamma level in the local femoral head of rabbits in Jiangu decoction group decreased compared with model group (P=0.033).

CONCLUSIONThe Jiangu decoction is effective to inhibit the femoral head adipogenic differentiation by decrease the PPAR content, so as to prevent and treat steroid-induced femoral head necrosis.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Femur Head Necrosis ; chemically induced ; drug therapy ; pathology ; Glucocorticoids ; blood ; toxicity ; Male ; Medicine, Chinese Traditional ; PPAR gamma ; analysis ; Rabbits

Background: and Purpose The relationships among interleukin (IL)-10 levels, anxiety, and cognitive status after stroke remain controversial. We aimed to determine the associations of serum IL-10 levels with poststroke anxiety (PSA) and poststroke cognitive impairment (PSCI). Methods: We recruited 350 patients with stroke, of whom only 151 completed a 1-month follow-up assessment. The Mini Mental State Examination (MMSE) and Hamilton Anxiety Scale (HAMA) were used to assess the cognitive status and anxiety, respectively. Serum IL-10 levels were measured within 24 hours of admission. Results: IL-10 levels were significantly lower in the PSA group than in the non-PSA group, and they were negatively associated with HAMA scores (r=-0.371, p<0.001). After adjusting for all potential confounders, IL-10 levels remained an independent predictor of PSA (odds ratio=0.471, 95% confidence interval=0.237–0.936, p=0.032). IL-10 levels were strongly correlated with behavior during interviews, psychic anxiety, and somatic anxiety. Patients without PSCI had higher IL-10 levels were higher in non-PSCI patients than in PSCI patients, and they were positively associated with MMSE scores in the bivariate correlation analysis (r=0.169, p=0.038), and also with memory capacity, naming ability, and copying capacity.However, IL-10 did not predict PSCI in the univariable or multivariable logistic regression. Conclusions Low IL-10 levels were associated with increased risks of PSA and PSCI at a 1-month follow-up after stroke. Serum IL-10 levels may therefore be helpful in predicting PSA.

OBJECTIVESTo review the preliminary clinical experience with high-field-strength intraoperative magnetic resonance imaging (iMRI) suite with neuronavigation system in the pituitary adenoma operation with transsphenoidal approach.

METHODSFrom March 2009 to December 2010, 31 patients [range, 29 - 76 years, mean age (47 ± 11) years]of pituitary adenoma were operated with transsphenoidal approach and intraoperatively with a movable 1.5 T high-field-strength iMRI suite in combination with neuronavigation system. Tumor size was 1.8 - 7.3 cm, mean (3.5 ± 1.2) cm. Twenty-five cases were non-functional pituitary adenoma, 4 cases were prolactin-secreting pituitary adenoma, 2 cases were growth hormone-secreting pituitary adenoma. Thirty patients' resection with transnasal transsphenoidal approach were performed, one patient with transoral transsphenoidal approach was performed.

RESULTSIn 12 cases of 30 patients who planed to totally remove tumor, iMRI had revealed residual lesions and resulted in the change of the surgical strategy, 2 invasive cavernous sinus cases no further resection of the tumor because of internal carotid artery encasement, the other 10 cases resected further, eventually. Finally, 8 cases were totally removed. The ratio of total removal tumor was enhanced to 86.7% (26/30) from 60.0% (18/30). There was no perioperative mortality.

CONCLUSIONSHigh-field-strength iMRI suite with neuronavigation system provides valuable information of tumor resection that allows intraoperative modification of the surgical strategy. It could be very helpful to maximize the resection of the pituitary adenoma and minimize the injury to neurological function.

Adenoma ; surgery ; Adult ; Aged ; Cavernous Sinus ; surgery ; Female ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Monitoring, Intraoperative ; methods ; Neuronavigation ; methods ; Pituitary Neoplasms ; surgery

Background: Genetic defects in the human thyroid-stimulating hormone (TSH) receptor (TSHR) gene can cause congenital hypothyroidism (CH). However, the biological functions and comprehensive genotype–phenotype relationships for most TSHR variants associated with CH remain unexplored. We aimed to identify TSHR variants in Chinese patients with CH, analyze the functions of the variants, and explore the relationships between TSHR genotypes and clinical phenotypes. Methods: In total, 367 patients with CH were recruited for TSHR variant screening using whole-exome sequencing. The effects of the variants were evaluated by in-silico programs such as SIFT and polyphen2. Furthermore, these variants were transfected into 293T cells to detect their Gs/cyclic AMP and Gq/11 signaling activity. Results: Among the 367 patients with CH, 17 TSHR variants, including three novel variants, were identified in 45 patients, and 18 patients carried biallelic TSHR variants. In vitro experiments showed that 10 variants were associated with Gs/cyclic AMP and Gq/11 signaling pathway impairment to varying degrees. Patients with TSHR biallelic variants had lower serum TSH levels and higher free triiodothyronine and thyroxine levels at diagnosis than those with DUOX2 biallelic variants. Conclusions We found a high frequency of TSHR variants in Chinese patients with CH (12.3%), and 4.9% of cases were caused by TSHR biallelic variants. Ten variants were identified as loss-of-function variants. The data suggest that the clinical phenotype of CH patients caused by TSHR biallelic variants is relatively mild. Our study expands the TSHR variant spectrum and provides further evidence for the elucidation of the genetic etiology of CH.

Objective To explore the prognostic factors of central venous catheter-related bloodstream infection(CR-BSI)and provide reference for clinical practice. Methods The clinical data of 346 CR-BSI patients from February 2014 to July 2019 were retrospectively reviewed,and the prognostic factors were analyzed. Results Of the 346 CR-BSI patients,62 died,yielding a case-fatality rate of 17.92%.Univariate analysis showed that 18 factors including age(
Anti-Bacterial Agents ; Carbapenem-Resistant Enterobacteriaceae ; Central Venous Catheters/adverse effects* ; Humans ; Hyperglycemia ; Hypoproteinemia ; Klebsiella Infections ; Klebsiella pneumoniae ; Methicillin-Resistant Staphylococcus aureus ; Mycoses ; Prognosis ; Pseudomonas Infections ; Retrospective Studies ; Risk Factors ; Sepsis/mortality*

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