Objective To observe the effects of folic acid(FA)supplementation on the expression of Flotillin-1 and β-amyloid protein(Aβ)-metabolism-related proteins in the brains of inflammation-stimulated Alzheimer's disease(AD)mice.Methods Twenty-seven 6-month-old male APP/PS1 mice were randomly divided into AD,AD+LPS,and AD+LPS+FA groups,with nine mice in each group.Nine C57BL/6J male mice born within the same month were used as the Control group.The AD+LPS+FA group was given folic-acid-supplemented feed(8 mg/kg)for 3 months of intervention,while the other three groups were fed normal feed.Lipopolysaccharide solution(LPS,250 μg/(kg·d))was injected intraperitoneally into mice in the AD+LPS and AD+LPS+FA groups 1 week before the end of the experiment,and saline was injected into the remaining two groups.The serum inflammatory factors TNF-α and IL-6 levels and brain tissue Aβ1-40 and Aβ1-42 levels of mice in each group were detected by ELISA.Flotillin-1 protein expression in brain tissue was detected using Western blot,and the co-expression of Flotillin-1 and Aβ1-42/APP/PS1/BACE1 in the cortical region of the brain was detected via immunofluorescence double-labeling.Results After ANOVA analysis,we found mice in the AD group had elevated serum TNF-α and IL-6 levels(P＜0.05),elevated levels of Aβ1-40 and Aβ1-42(P＜0.05),increased expression of Flotillin-1 protein(P＜0.05),and increased co-expression of Flotillin-1 and Aβ1-42/APP/PS1/BACE1 in the cortical brain tissue(P＜0.05)compared with the Control group.Compared with mice in the AD group,those in the AD+LPS group had further increases in serum inflammatory factors and Aβ levels in the brain(P＜0.05)and increased co-expression of Flotillin-1 and Aβ1-42/APP/BACE1 double-labeled proteins in their cortical brain tissue(P＜0.05).Compared with mice in the AD+LPS group,those in the AD+LPS+FA group had lower in vivo inflammation levels and Aβ content in the brain(P＜0.05),lower brain tissue Flotillin-1 protein expression(P＜0.05),and lower Flotillin-1 and Aβ1-42/APP/PS1/BACE1 protein co-expression in cortical brain tissue(P＜0.05).Conclusions Folic acid supplementation may reduce Flotillin-1 protein expression and Aβ deposition in the brain of AD inflammatory mice.

Objective:To evaluate the effect of extracorporeal shock wave on the phosphoproteomics of the spinal cord in rats with diabetic neuralgia.Methods:Thirty-six healthy male SPF-grade Sprague-Dawley rats, aged 2 months, weighing 200-250 g, were divided into 3 groups ( n=12 each) using the random number table method: control group (group C), diabetic neuralgia group (group D), and extracorporeal shock wave + diabetic neuralgia group (group E). The rats were continuously fed a common diet in group C, while the rats were fed a high-sugar and high-fat diet for 8 weeks in D and E groups. Streptozotocin 35 mg/kg was intraperitoneally injected, and the successful induction of diabetic neuralgia was defined as the blood glucose >14.6 mmol/L and the mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) ≤85% of baseline values. Group E received extracorporeal shock wave treatment after developing the model, with 1, 000 shocks per session at a frequency of 10 Hz and an energy of 1.0 bar, once per week for a total of 4 sessions. The MWT and TWL were measured before developing the model (T 0) and at 1, 2, 3 and 4 weeks after developing the model (T 1-T 4). After the last extracorporeal shock wave treatment, the rats were anesthetized and sacrificed, and lumbar spinal cord tissues were obtained for proteomic analysis and for detection of the expression of glial fibrillary acidic protein (GFAP), interleukin-1beta (IL-1β), and tumor necrosis factor-alpha (TNF-α) (by immunohistochemistry). Results:Compared with group C, the MWT and TWL were significantly decreased at T 1-T 4 in D and E groups ( P<0.05). Compared with group D, the MWT and TWL were significantly increased at T 1-T 4 in group E ( P<0.05). The results of phosphoproteomics screening revealed 284 differentially phosphorylated proteins in D and C groups, 282 in E and C groups, and 303 in E and D groups ( P<0.05). The results of immunohistochemistry showed that the expression of GFAP, IL-1β and TNF-α was significantly up-regulated in group D compared with group C ( P<0.05); the expression of GFAP, IL-1β and TNF-α was significantly down-regulated in group E compared with group D ( P<0.05). Conclusions:The mechanism by which extracorporeal shock wave alleviates diabetic neuralgia is related to inhibition of astrocyte activation and excessive phosphorylation of mGluR5 in rats.

Objective:To study the clinical features and treatment strategy of neonatal ureaplasma meningitis.Methods:During 2021, the clinical data of 2 neonates with ureaplasma meningitis treated in Children's Hospital of Hunan Province were retrospectively analyzed. Literature on this subject were searched in the following databases: CNKI, Wanfang Database, Chinese Medical Journal Full-Text Database, CQVIP database, SinoMed, PubMed, Embase and Web of Science (up to March 2022). The key words included “infant”, “neonate”, “newborn”, “ureaplasma”, “mycoplasma urealytium”, “meningitis”, “central nervous system infection”, “brain”. The clinical data, treatment and prognosis of patients from the literature were summarized.Results:Case 1, female, gestational age(GA) 33 +3 weeks, intracranial hemorrhage (ICH) and ventricular dilatation were found on 2 d after birth. The cerebrospinal fluid (CSF) routine and biochemistry tests indicated meningitis, but the CSF culture was negative. No improvement after antibiotic treatment. CSF metagenomic next-generation sequencing (mNGS) and 23S rRNA showed Ureaplasma urealyticum on 30 d after birth. The patient was treated with doxycycline (DOX) for 21 d until mNGS turned negative and DOX was discontinued. However, the disease recurred 23 d later and erythromycin was added with DOX as combined therapy. The patient was followed up until 6 months without neurodevelopmental disabilities. Case 2, male, GA 26 weeks, ICH and ventricular dilatation were found on 10 d after birth. The CSF routine and biochemistry tests indicated meningitis, but the CSF culture was negative. No improvement after antibiotic treatment. CSF mNGS and 23S rRNA showed Ureaplasma parvum. The patient received erythromycin therapy for 32 d and had normal neurodevelopment at 5 months. According to the literature, 43 cases were reported including the 2 cases descirbed above, 17 cases were full-term infants and 26 cases were preterm infants. The median CSF leukocytes, glucose and proteins were 566 cells/mm 3, 0.2 mmol/L and 2.2 g/L. 27 cases were diagnosed based on CSF culture, 6 cases using mNGS, 4 cases with both CSF culture and PCR method and 6 cases with other methods. Macrolides alone were used in 14 cases, macrolides combined with another antibiotic were used in 8 cases, non-macrolide antibiotics were used in 9 cases and 12 cases didn't receive any anti-ureaplasma therapy. All 17 term infants survived, however, 8 cases with hydrocephalus. Among the 26 preterm infants, 8 patients died, 18 patients had periventricular-intraventricular hemorrhage and 15 patients had hydrocephalus. Conclusions:Neonatal ureaplasma meningitis has significantly lower CSF glucose level with hydrocephalus as the common complication. For intracranial infections of unknown etiology and no response to treatment, mNGS is helpful in determining the pathogen.Neonatal ureaplasma meningitis should be treated with macrolides alone or as add-on therapy.

Objective:To evaluate the effect of intravenous infusion of lidocaine on the efficacy of conventional treatment for rheumatoid arthritis.Methods:Forty-four patients with rheumatoid arthritis of either sex, aged 32-85 yr, weighing 40-76 kg, who were admitted to the Department of Pain and Nephrology in our hospital from September 2019 to September 2020, were divided into 2 groups ( n=22 each) according to the random number table method: control group (C group) and lidocaine group (L group). Both groups received conventional treatment.When visual analogue scale (VAS) score ≥5, glucocorticoid (GC) and non-steroidal anti-inflammatory drugs (NSAIDs) were taken orally to maintain the VAS score ≤4.In group L, 0.2% lidocaine hydrochloride injection 3 mg/kg (diluted with 0.9% sodium chloride injection 500 ml) was intravenously infused at a rate of 25 ml/h for 2 h, once a day, for 5 consecutive days, based on the conventional treatment.The VAS score, 28-joint Disease Activity Score (DAS28 score), simplified disease activity index score (SDAI score), consumption of GC and NSAIDs and adverse reactions were recorded before treatment (T 1) and at 1, 4 and 8 weeks after treatment (T 2-4). The temperature of the pain area of the affected joint was evaluated through infrared thermal imaging at T 1 and T 2. Results:Compared with the baseline at T 1, VAS score, DAS28 score and SDAI score were significantly decreased at each time point, and the temperature of the pain area of the affected joint at T 2 was decreased in the two groups ( P<0.05). There were no significant differences in VAS score, DAS28 score and SDAI score at each time point between two groups ( P>0.05). Compared with group C, the consumption of GC and NSAIDs was significantly decreased, and the temperature of the pain area of the dorsum of both hands and the dorsum of right foot at T 2 and incidence of adverse reactions were decreased in group L ( P<0.05). Conclusions:Intravenous infusion of lidocaine can optimize the efficacy of conventional treatment for rheumatoid arthritis.

Objective:To develop a self-made plasma quality control material for non-invasive prenatal testing (NIPT) and evaluate its performance.Methods:139 NIPT-negative maternal plasmas stored in the genetic department of Shaoxing maternal and child health hospital from January 1, 2019 to June 30, 2021 were divided into male groups (19 cases) and female groups (120 cases) according to the neonatal gender. 9360 cases from September 2020 to September 2021 were enrolled as clinical validation cases.First step, 200 μl plasma from a 47 years-old non-pregnant healthy women was used as a matrix. Different amounts (0.1, 0.2, 0.5, 2.5, and 5 μl) of positive DNA from fetal chromosome aneuploidy (T21, T18, T13) detection kit were added. The appropriate volume of positive DNA was 0.5 μl according to the test results. Second step,Plasma in male and female group was treated as matrix. 0.5 μl positive DNA was added per 205 μl. Plasma matrix from female group showed good repeatability and the sensitivity was 100%.Third step, evaluate the self-made plasma quality control material, including storage stability, matrix uniformity and repeatability, and the effect of different batch numbers of positive DNA, by calculating Z score and the CV of fetal DNA concentration (FF).Results:Plasma matrix from female group showed good repeatability and the sensitivity was 100%, while the sensitivity of male group was only 84%. The CV of FF in female matrix was 3.9% in the repetitive experiments. After adding 0.5 μl positive DNA, the mean FF of self-made positive plasma quality control was 5.63%±0.42%, Z values>6, and the CV was 7% after storage of three months. Considering the concentration variation of positive DNA in different lots, 1 μl of positive DNA should be added when the FF of positive DNA is lower than 10%.Used in 9360 clinical cases from September 2020 to September 2021, all positive plasma quality control materials showed positive results, and the positive predictive value of trisomy 21 was 100%.Conclusions:The NIPT self-made positive plasma quality control material has been successfully developed in this study. The preliminary experimental results show that it has good repeatability and stability, which is suitable for clinical application.

Objective:To investigate the dosimetry differences in the treatment of locally advanced cervical cancer with intracavitary/interstitial brachytherapy (IC/IS BT), intracavity brachytherapy combined with intensity modulated radiotherapy (ICBT+ IMRT) and simple IMRT.Methods:Totally 16 patients with local advanced cervical cancer were retrospectively selected, which were treated by three-dimensional brachytherapy. On the basis of the original three-dimensional intracavitary/interstitial brachytherapy plan, ICBT+ IMRT and IMRT plans were designed respectively to study the dosimetry differences of target and different organs at risk for the three kinds of plans.Results:A total of 75 brachytherapy treatment plans were designed, including 25 IC/IS BT, 25 ICBT+ IMRT and 25 IMRT. There was not statistically significant difference of target dose parameters between ICBT+ IMRT and IC/IS BT plan ( P>0.05). ICBT+ IMRT plans had better OAR sparing than IC/IS BT. The doses of OARs in the IMRT plans were relatively large and the volume irradiated to more than 60 Gy ( V60) was significantly higher( t=6.77, 10.37, 4.61, 2.83, P<0.05). Conclusions:The ICBT+ IMRT technique not only provides better target coverage, but also maintains low doses to the OARS, which can be used as an alternative treatment to IC/IS BT. Although the target coverage of IMRT is better, the protection of OARs is not satisfied, so it is not suitable for local boost therapy of advanced cervical cancer.

Objective To investigate the early growth and development of extremely low birth weight infants (ELBWI) and very low birth weight infants (VLBWI) through a follow-up study from hospital discharge until 18 months of corrected age.Methods ELBWI and VLBWI who were hospitalized and discharged alive from the Neonatal Intensive Care Unit of Hunan Children's Hospital from January 2013 to June 2014 were recruited.Follow-ups were performed at the corrected age of 40 weeks,as well as at one,three,six,12 and 18 months of corrected age.Several parameters indicating the growth and development of those infants were monitored and assessed.Extrauterine growth retardation (EUGR) was defined as head circumference (HC) or weight ≤ 10th percentile for gestational age at discharge.T-,rank-sum,or Chi-square (or Fisher's exact) test was performed for statistical analysis.Results (1) A total of 285 ELBWI and VLBWI were recruited.Among them,145 (50.9％) were alive at last follow-up,37 (13.0％) died,and 103 (36.1％) were lost.No significant differences in clinical data were observed between the infants who completed the follow-up and those who did not (all P＞0.05).(2) Based on HC and weight,the incidences of EUGR in the 145 infants reached the peak at the corrected age of three months [42.8％ (62/145) and 40.0％ (58/145)],and then declined with increasing age.At 18 months of corrected age,the incidences of EUGR dropped to 31.7％ (46/145) and 14.5％ (21/145),respectively.(3) There were no significant differences in gender,gestational age,birth weight,length of hospital stay,duration of oxygen therapy,and incidences of complications between the infants with and without EUGR (allP＞0.05).(4) The rate of pulmonary surfactant therapy in neonates with EUGR was lower than in those without [27.8％ (15/54) vs 53.8％ (49/91),x2=9.340,P＜0.05].There were no significant differences in mental development index and psycho-motor development index at 12 and 18 months of corrected age between the neonates with and without EUGR (all P＞0.05).Neither HC nor weight at the corrected age of 18 months showed significant differences between the two groups (both P＞0.05).(5) At 18 months of corrected age,31.7％ (46/145) of the infants had their HC ≤ 10th percentile,and 14.5％ (21/145) had their weight ≤ 10th percentile.Infants with HC ≤ 10th percentile were at higher risk of abnormal neurodevelopment than those with HC ＞10th percentile [67.4％ (31/45) vs 40.4％ (40/99),X=9.154].Infants with either HC or weight ≤ 10th percentile had higher risk of abnormal neurodevelopment that those with both HC and weight ＞10th percentile [65.5％ (36/55) vs 38.9％ (35/90),x2=9.641] (both P＞0.05).Conclusions ELBWI/VLBWI are at high risk of growth retardation.Incidence of growth restriction declines with age.

Objective To analysis the clinical outcome at discharge and its risk factors of extremely preterm infants.Method To retrospectively analysis the clinical outcome at discharge and it's risk factors of extremely preterm infants (less than 28 weeks gestation) admitted from September 2008 to August 2014 in our Hospital.Result A total of 179 cases were enrolled.Survival rate was 59.2％ (106/179).Unfavorable outcome rate was 74.3％ (133/179),among them 73 cases died.The top five causes of death were severe bronchopulmonary dysplasia (BPD) (28 cases),Ⅲ ～ Ⅳ o intraventricular hemorrhage (IVH) (19 cases),sepsis (16 cases) and necrotizing enterocolitis (NEC) (6 cases).Among the 60 survivals with unfavorable outcomes,35 cases had either severe neurologic or ophthalmological sequela,and 25 cases had severe pulmonary sequela.Univariate analysis showed that,comparing with improved group,unfavorable outcome group had higher rates of not receiving prenatal steroids,placental abruption,male,small for gestation age,resuscitation with chest compression,admission age older than 72 hour,severe respiratory distress syndrome (RDS),without pulmonary surfactant (PS) usage,mechanical ventilation beyond 2 weeks and sepsis (P ＜ 0.05).Logistic regression analysis showed that those without prenatal steroids (OR =9.402,P =0.002),small for gestational age (OR =8.271,P =0.018),resuscitation with chest compression (OR =6.325,P =0.023),admission age older than 72 hour (OR =4.174,P =0.028) were independent risk factors for unfavorable outcome of extremely premature at discharge.Conclusion Extremely preterm infants have a higher rate of unfavorable outcome at discharge.Avoid small for gestational age,transfer properly and in time both in utero and after birth,and conduct prenatal steroids could improve their clinical outcome at discharge.

Objective To explore the clinical effect of the radial artery perforator flap on repairing soft tissue defect after palm postoperative scar contracture. Methods Eighteen patients with palm scar contracture were selected. The palm soft tissue defect was repaired by radial artery perforator flap after the operation of scar removal and soft tissue release. The areas of soft tissue ranged from 5 cm × 3 cm to 8 cm × 5 cm. The area of donor flap exceeds 20%of the wounds area, and wounds were repaired by free skin grafting. Six months after operation, the hand function was compared with that before operation. Results All the patients were followed up for 6-15 months (mean 10.8 months), all flaps survived with good shape, and flap donor site wounds were healed by skin grafting. The patients were evaluated 6 months after operation according to the Chinese Medical Hand Surgery Society of upper part of functional assessment criteria: excellent in 11 cases, good in 5 cases and general in 2 cases, but preoperative functional evaluation was excellent 0 case, good in 3 cases, general in 7 cases and poor in 8 cases. Compared with that before operation, the postoperative function was significantly improved. Conclusions It is a commendable approach of repairing soft tissue defect after postoperative palm scar contracture by radial artery perforator flap, because it can provide reliable blood supply, and significantly improve hand function with exactly clinical effect.

Objective:To investigate the role of epidermal growth factor receptor (EGFR)in the regulation of B[a]P-induced silent information regulator 1 (SIRT1 ) activation in the human bronchial epithelial cells (BEAS-2B),and to clarify the relationship between EGFR/SIRT1 signal transduction pathway and the occurrence and development lung cancer.Methods:The prediction analysis of transcriptional factor binding sites of SIRT1 was performed by MOTIF SearchTM software.The primary cultivated BEAS-2B cells were divided into control group (without B[a]P exposure)and B[a]P groups (the cells were exposed to B[a]P for 6,12,24,48 h).RT-PCR and Western blotting method were carried out to detect the EGFR mRNA and protein expression levels.The BEAS-2B cells transfected with SIRT1 promotor luciferase reporter gene plasmid were treated with human epidermal growth factor (hEGF)and Genistein (tyrosine protein kinase inhibitor)for 24 h,the morphology of BEAS-2B cells was observed by inverted microscope, and luciferase reporter assay was used to test the SIRT1 transcriptional activity.The human lung tissue biopsies were acquired and the immunohistochemical analysis was used to determine the EGFR protein expression level.Results:The transcriptional factor binding sites of SIRT1 contained EGF, 2Fe-2S and vWF.Compared with control group,the expression levels of EGFR mRNA in B [a]P groups were increased in a time-dependent manner,and reached the peak at 12 h (P < 0.05);the EGFR protein expression levels were also increased (P <0.05).The SIRT1 luciferase activity in hEGF group was increased compared with control group (P <0.05);when hEGF and B[a]P worked together,the SIRT1 luciferase activity was increased even further (P <0.001). The cells showed arrangement and morphologic changes gradually when the B[a]P concentration was above 30 μmol · L-1. Genistein (30 μmol · L-1 )inhibited the increase of SIRT1 luciferase activity induced by B [a]P ,and there was significant difference compared with control group (P <0.05).The immunohistochemistry results showed that EGFR expression level in lung cancer tissue was higher than that in normal lung tissue (P < 0.001).Conclusion:EGFR can regulate the B [a]P-induced SIRT1 expression in BEAS-2B cells,and to cause lung chronic inflammation;EGFR/SIRT1 signal transduction pathway may play a role in the occurrence and development of lung cancer.