Abstract: Bartter syndrome (BS) is a kind of inherited metabolic disease characterized by electrolyte and endocrine disorder, resulting from genetic gene mutation or deletion. Clinically, it manifests as vomiting, constipation, feeding difficulties, weight loss, growth retardation. The laboratory examination shows hypokalemia, metabolic alkalosis, hyperreninemia, aldosteronism, etc. As an autosomal recessive genetic disease, BS has an extremely low incidence rate, making diagnosis and treatment quite challenging. In recent years, with the progress of gene detection and other technologies, China has made great progress in the study of BS, more and more BS has been diagnosed accurately. According to known gene mutation types, it can be divided into type 1, type 2, type 3, type 4, type 5 and Gitelman syndrome, a total of 6 types, with type 4 further divided into type 4a and type 4b. At present, the most common type of children with BS in clinical practice is type 3, which is the classic type of BS. This paper reports a case of Bartter syndrome type 4b (BS4b). After two rounds of three-generation family gene sequencing, it was discovered that the infant had a combined mutation of both the CLCNKA and CLCNKB alleles, in addition to Alport syndrome, and both parents were carriers of this type of gene defect. The child was finally diagnosed as BS4b and Alport syndrome. This report, combined with the patient's clinical features, diagnosis and treatment process, as well as related literature analysis, aims to provide experience for understanding and diagnosis and treatment of this disease.

Objective : To investigate whether the application of melatonin (MT) in embryo culture in vitro can improve the treatment effect of in vitro fertilization embryo transfer(IVF⁃ET) in patients with decreasing ovarian reserve (DOR) . Methods : 128 DOR patients receiving assisted reproductive therapy were collected. All patients were treated with an antagonist scheme of super⁃ovulation. Patients were divided into melatonin group (n = 56) and control group (n = 72) according to whether melatonin ( melatonin concentration 10 - 9 mol/L) was added into embryo culture medium. Results : There was no statistically significant difference in oocytes fertilization rate and cleavage rate between the two groups during later embryo culture , but blastocyst formation rate ( 65. 22% vs 56. 16% ) and high⁃quality blastocyst rate (52. 96% vs 40. 94% ) in the melatonin group were higher than those in the control group , and the differences were statistically significant ( P < 0. 05 ) . There were no significant differences in the implantation rate (50. 00% vs 38. 67% ) and clinical pregnancy rate (48. 39% vs 46. 00% ) of blastocysts after freezing⁃thawing between the two groups , but the cycle number of high⁃quality blastocysts obtained in the melatonin group was higher than that in the control group (85. 71% vs 69. 44% ) , and the difference was statistically significant (P < 0. 05) . Conclusion In a way , the application of melatonin in the in vitro culture of early embryos can promote the development of oocytes in patients with DOR , improve the quality of embryos , and finally substantially improve the therapeutic effect of such patients.

Objective: To explore the effects of melatonin(MT) on the fertilization of in vitro matured oocytes in controlled ovarian hyperstimulation(COH) cycles and the development of subsequent embryos. Methods: Imma-ture human oocytes from COH cycles were collected for in vitro maturation(IVM) culture and intracytoplasmic sperm injection( ICSI) insemination,and then the cleavage embryo and blastocyst culture medium supplemented with 0,10-11,10-9,10-7or 10-5mol/L MT were used in order to perform embryo culture in vitro,next the formed high-quality blastocysts were picked up and cryopreserved via vitrification,finally,array-CGH technology was used to detect aneuploidy of rewarmed high-quality blastocysts. Results: The blastocyst rate of 10-9mol/L group was significantly higher than that of 10-11,10-7,10-5and 0 mol/L groups( P<0. 05,P<0. 01,P<0. 000 1,P<0. 01),respectively; the high-quality blastocyst rate of 10-9mol/L group was higher than that of other groups,but there were only significant differences compared with 10-5mol/L and 0 mol/L groups( P<0. 01,P<0. 05); the incidence of aneuploidy in MT group( 17. 6%) was lower than that in non-MT group( 33. 3%),but no significant difference was found. Conclusion The addition of MT to human embryo culture medium can promote the in vitro development of embryo,which is related to concentration,and 10-9mol/L is the optimal concentration.

Objective:To detective the cut-off values of amino acid levels in premature infants in Sichuan.Methods:Data of newborns screening for inherited metabolic diseases (IMD) by tandem mass spectrometry in Sichuan Province from January 2018 to December 2019 were retrospectively analyzed.They were divided into premature infant group ( n=2 264, 1 312 males and 952 females) and full-term infant group ( n=53 275, 28 269 males and 25 006 females). The cut-off values of amino acids in dry blood spots were expressed as percentage ( P0.5 - P99.5), and rank sum test was used for comparison between preterm and full-term infants. Results:(1) The distribution of 11 amino acids [alanine (ALA), arginine (ARG), citrulline (CIT), glycine(GLY), leucine (LEU), methionine (MET), ornithine (ORN), phenylalanine (PHE), proline (PRO), tyrosine (TYR) and valine (VAL)] in premature infants were abnormal.(2) The cut-off values of amino acids in premature infants were as follows: ALA: 135.20-552.33 μmol/L, ARG: 1.34-47.04 μmol/L, CIT: 5.66-32.02 μmol/L, GLY: 181.48-909.93 μmol/L, LEU : 71.10-283.29 μmol/L, MET: 4.21-34.51 μmol/L, ORN: 40.58-293.76 μmol/L, PHE: 23.60-106.30 μmol/L, PRO: 77.76-358.24 μmol/L, TYR: 27.52-352.91 μmol/L, VAL: 53.74-228.37 μmol/L.(3) The cut-off values of amino acid in full-term infants were as follows: ALA: 135.20-552.33 μmol/L, ARG: 1.30-42.73 μmol/L, CIT: 5.92-30.35 μmol/L, GLY: 208.17-980.09 μmol/L, LEU: 72.91-287.49 μmol/L, MET: 4.27-33.90 μmol/L, ORN: 48.40-305.59 μmol/L, PHE: 27.63-92.27 μmol/L, PRO: 97.38-372.75 μmol/L, TYR: 40.19-276.54 μmol/L, VAL: 65.75-237.92 μmol/L.(4) Except for PHE ( Z=-0.58, P>0.05), the other indicators were significantly different between 2 groups [ALA ( Z=-15.32, P<0.05), ARG ( Z=-5.62, P<0.05), CIT ( Z=-5.86, P<0.05), GLY ( Z=-14.52, P<0.05), LEU ( Z=-5.62, P<0.05), MET ( Z=-5.22, P<0.05), ORN ( Z=-13.01, P<0.05), PRO ( Z=-22.09, P<0.05), TRY ( Z=-2.09, P<0.05), VAL ( Z=-17.82, P<0.05)]. Conclusions:The establishment of the cut-off values of amino acids in premature infants in Sichuan provides a theoretical basis for laboratory diagnosis of IMD screening, which enhances the accuracy of diagnosis and avoids excessive medical treatment.

Background Neck-shoulder pain is one of the most common discomfort symptoms among nursing staff, mostly caused by a heavy workload, restricted workstation, and prolonged poor posture. Objective To investigate the prevalence of neck-shoulder pain among nursing staff in tertiary general hospitals in Hunan Province, and to analyze workload-related risk factors. Methods From October to December 2018, a multi-stage stratified randomized cluster sampling method was adopted to select a total of 1 200 nursing staff who met the inclusion andexclusion criteria in six tertiary general hospitals from five geographic subdivisions of Hunan Province were selected. A self-designed questionnaire was used to collect demographic characteristics, and past 1-month prevalence of neck-shoulder pain, a neck disability index (NDI) was used to assess the impact of neck pain on daily life, and functional disability was graded by the index of impaired neck function; a self-designed questionnaire was adopted to evaluate potential workload-related risk factors (working time, postural load, and force load) of neck-shoulder pain after a Delphi expert consultation. A total of 1 161 valid questionnaires were returned, and χ^２test and logistic regression model were used to screen the potential risk factors for neck-shoulder pain. Results The prevalence rate of neck-shoulder pain in the past 1 month was 87.5% (1 016/1 161) and varied by different characteristics of the nursing staff, ranging from 76.9% to 91.2%. The scores of the 10 items of the NDI ranged from 0 to 5, but most of them were 0, 1, and 2; of the 10 items, neck pain had the greatest impact on sleep (1.30±1.21) and the least impact on self-care behaviors (0.35±0.65). The index of impaired neck function showed that the majority (65.4%) of nurses had mild cervical spine dysfunction and 30.1% had moderate cervical spine dysfunction. The univariate analysis results revealed that 24 out of the 31 workload-related risk factors had significant differences, and further multiple analysis results showed that three variables entered the logistic regression model. Under the same conditions, nurses with ≥5 h of cumulative head-down tasks per shift had 3.03 times higher neck-shoulder pain risks compared with those with ＜1 h. Compared with “occasionally or never”, nurses who “sometimes” and “often” tilted their necks back and maintained this posture for a long time showed 2.36 and 2.94 times higher risks for neck-shoulder pain respectively. The nurses who reported “sometimes” and “often” having difficulty using their force at work due to unnatural posture had 2.78 times and 7.08 times higher neck-shoulder pain risks than those who “occasionally or never” respectively. Conclusion The reported rate of neck-shoulder pain among nurses in tertiary general hospitals in Hunan Province is high, but most of them are mild dysfunction. Working hours and posture load may affect the risk of neck-shoulder pain.

Objective:To explore the role of hippocampal γ oscillation abnormality in sepsis-associated encephalopathy (SAE).Methods:Seventy male Sprague-Dawley rats (2-3 months) were randomly (random number) divided into three groups according to the random digital table method: sham, CLP, and CLP + dopamine 4 (D4) receptor agonists RO-10-5824 group. The SAE animal model was established by cecal ligation and puncture (CLP). On day 10-14 after surgery, the open field, novel object recognition, and fear conditioning tests were performed. After that, the hippocampus was collected to measure expressions of parvalbumin (PV) and D4 receptor. In another set of experiment, CA1 local field potential (LFP) were recorded, and the relationship between LFP and time with novel object was analyzed. Independent sample t-test was used for pairwise comparisons, and multiple comparisons were performed by one-way ANOVA, followed by the Tukey multiple comparisons test. Correlation was analyzed using Pearson correlation. Statistical significance was assumed when P<0.05. Results:Compared with the sham group, hippocampal PV (77.54±4.61)%, D4 expression (56.36±3.88)% and γ oscillation power (41.1±8.62)%, object exposure time (36±3) s, new object recognition rate (49±4)%, and scene stiffness time (56±7) s were decreased significantly ( P<0.05). However, RO-10-5824 treatment could increase hippocaml γ oscillation power (92.3±6.7)%, and reverse the decreased new object exposure time (44±3) s and new object recognition rate (63±4)%. Correlation analysis showed that hippocampal γ oscillation power was positively associated with new object exposure time ( r=0.609 2, P=0.015 9). There was no difference in total distance traveled or time spent in the center among groups ( P>0.05). Conclusion:Hippocampal γ oscillation abnormality might play a key role in cognitive impairment associated with SAE.

Objective:To explore the change characteristics of amino acid levels in neonates, so as to provide theoretical basis for accurate clinical interpretation.Methods:By preliminary screening and diagnosis from 32 855 newborns, 32 843 samples were collected using tandem mass spectrometry to inherited metabolic disease (IMD) scree-ning in Sichuan Province from January to December 2018.Afterwards, according to gestational age (1 363 premature infants, 31 468 full-term infants and 12 overdue infants), blood collection time (3-7 days old, 3 095 cases, 8-28 days old, 1 637 cases, and more than 28 days old, 248 cases) and season (7 737 cases in the first quarter, 11 428 cases in the second quarter, 5 482 cases in the third quarter, and 8 196 cases in the fourth quarter), neonates were divided into different study groups.The difference of amino acid level in each group was compared, and the correlation between various influencing factors and metabolic index was analyzed.Results:(1) The distribution of 11 amino acids [alanine(ALA), arginine(ARG), citrulline(CIT), glycine(GLY), leucine+ isoleucine+ hydroxyproline (LEU+ ILE+ PRO-OH), methionine(MET), ornithine(ORN), phenylalanine(PHE), proline(PRO), tyrosine(TYR), and valine(VAL)] in neonates showed non-normally distribution.(2)The distribution of 11 amino acids in different gestational age were tested by nonparametric test, except for PHE( H=0.61, P>0.05)and TYR( H=2.02, P>0.05), and other indicators were significantly different [ALA( H=187.11, P<0.05), ARG( H=23.60, P<0.05), CIT( H=22.90, P<0.05), GLY( H=85.18, P<0.05), LEU( H=56.42, P<0.05), MET( H=18.74, P<0.05), ORN( H=129.27, P<0.05), PRO( H=344.40, P<0.05), and VAL( H=272.92, P<0.05)]. (3) The distribution of 11 amino acids in different blood collection time were significantly different [ALA( H=65.19, P<0.05), ARG( H=404.48, P<0.05), CIT( H=502.13, P<0.05), GLY( H=1 719.44, P<0.05), LEU( H=396.41, P<0.05), MET( H=199.39, P<0.05), ORN( H=31.26, P<0.05), PHE( H=325.49, P<0.05), PRO( H=70.09, P<0.05), TYR( H=159.29, P<0.05), and VAL( H=102.52, P<0.05)]. (4) The distribution of 11 amino acids in different birth seasons were significantly different [ALA( H=401.37, P<0.05), ARG( H=3 229.03, P<0.05), CIT( H=65.45, P<0.05), GLY( H=597.82, P<0.05), LEU( H=1 120.42, P<0.05), MET( H=10 515.18, P<0.05), ORN( H=1 275.23, P<0.05), PHE( H=2 260.17, P<0.05), PRO( H=319.57, P<0.05), TYR( H=884.37, P<0.05), and VAL( H=1 824.49, P<0.05)]. Conclusion:According to different gestational age, season and blood collection time, the metabolism of amino acids in neonates was different.When using tandem mass spectrometry for detection, appropriate interpretation criteria should be selected based on different conditions.

Objective:To evaluate the application of computer-assisted design and three-dimensional printing technique in reconstruction of maxillary defects with individual free fibular flap.Methods:A total of 13 patients, 7 males and 6 females with age from 12 to 55 years old, underwent the reconstruction of maxillary defects after subtotal or total maxillectomy for benign or malignant tumors between January 2016 and December 2018 were reviewed. Ther were 4 cases of subtotal maxillectomy and 9 cases of total maxillectomy. Before operation, osteotomy line was planned on three-dimensional images. A three-dimensional individual resin fibula model based on mirror images of the healthy side maxilla was obtained to fabricate an anatomically adapted osteomyocutaneous fibula free flap using computer-assisted design and forming. Oral, nasal, and eye functions and facial appearance were evaluated.Results:The 13 cases were followed up for 5-40 months, all flaps were alive except one due to flap failure and all cases were tumor free confirmed by CT or MRI. Ten patients could eat normal food without nasal food return, and 3 cases had palatal fistula. Eleven cases showed good speaking functional results. All cases had normal eye positions, no cases with diplopia and loss of vision. With evaluation by VAS, an average score of 8 was obtained, and most of patients were satisfied with their postoperative facial appearances.Conclusion:Reconstruction of maxillary defects by free fibular flap designed by three-dimensional printing and computer-assisted technique not only restored oral, nasal and eye functions, but also showed satisfactory facial appearance.

Objective To investigate the efficacy of preemptive analgesiawith ultrasound-guided modified dorsal penile nerve block combined ketorolac tromethamine for circumcision in pediatric patients.Methods Forty-five pediatric patients with ASA grade Ⅰ undergoing elective circumcision were randomly divided into 3 groups (n=15 each).The same intravenous compound anesthesia was used in the three groups.The pediatric patients underwent ultrasound-guided modified dorsal penile nerve block with a mixture of 0.1 ml/kg of 0.2% ropivacaine and 0.8% lidocaine before operation in group A and group B.The pediatric patients were given ketorolac tromethamine 1 mg/kg intramuscular injection before operation in group A and group C.The total consumption of propfol and sufentanil, occurrence of intraoperative body movement and respiratory depression, emergence time, time from waking up to going out of PACU and adverse reactions such as postoperative agitation, nausea, vomiting and pruritus were aslo recorded.The requirement for postoperative paracetamol suppositories was recorded.Results Compared with group C, the total consumption of propfol and sufentanil were significantly decreased, incidence of body movement andrespiratory depression were significantly decreased, the emergence time and time from waking up to going out of PACU was significantly shortened, requirement for paracetamol suppositories were significantly decreased in group A and group B(P<0.05).Compared with group B, incidence of requirement for paracetamol suppositories was significantly decreased in group A (P<0.05).No pediatric patients developed postoperative nausea, vomiting, pruritus and incidence of emergence agitation had no statistical difference in the three groups.Conclusion Preemptive analgesia with ultrasound-guided modified dorsal penile nerve block combined ketorolac tromethamine is safe and effective when used for circumcision in pediatric patients, and it has good efficacy of postoperative analgesia.

AIM:To investigate the effects of high plasma triglyceride (TG) caused by apolipoprotein C Ⅲ( ApoC Ⅲ) transgene on the occurrence and development of abdominal aortic aneurysm ( AAA) .METHODS:The animal models of hypercholesterolemia and hypercholesterolemia combined with hypertriglyceridemia were established by feeding high-fat diet to LDLR-/-and ApoC Ⅲ+LDLR-/-mice, respectively.AAA was induced in these mice by pancreatic elastase. By evaluating the incidence of AAA, relative maximal abdominal aortic diameter, disruption of the elastic lamellar structure and expression of matrix metalloproteinases ( MMPs) in the aorta walls of the AAA, the occurrence and development of AAA were compared in LDLR-/-and ApoC Ⅲ+LDLR-/-mice fed with either chow diet or high-fat diet.In addition, an in vitro TNF-α-induced aneurysmal microenvironment model on vascular smooth muscle cells ( VSMC) was used to study the impact of triglyceride-rich lipoproteins ( TRLs) from mice with normal or high contents of ApoCⅢon elastin protein expres-sion.RESULTS:Feeding the high-fat diet aggravated the severity of AAA in the LDLR-/-mice.ApoC Ⅲ+LDLR-/-mice fed with high-fat diet had less severe AAA than LDLR-/-mice fed with high-fat diet.TRLs inhibited degradation of VSMC elas-tin protein induced by TNF-α, and in vitro TRLs from the mice with high content of ApoC Ⅲ, compared to those with nor-mal content of ApoC Ⅲ, had better inhibitory effect on the degradation of elastin.CONCLUSION:High plasma TG caused by ApoC Ⅲtransgene alleviates AAA of the LDLR-/-mice induced by elastase and high-fat diet.The effect is probably attrib-uted to the hypertriglyceridemia caused by ApoC Ⅲtransgene.