Objecave To develop a sensitive and specific LC-MS/MS method for determination of testolactone in human urine.Methods A C_(18 )column(2.1×50mm,3.5μm) was used.The mobile phase Was a mixture of acetonitrile and the buffer solution(ammonium acetate-water solution adjusted with formic acid to pH 3.5)at a flow rate of 0.5ml/min.A mass spectrometer equipped with electrospray ionization source was used as a detector and operated in the positive mode.In multiple reaction monitoring(MRM)mode,the ion transitions of m/z 301→121 and m/z 301→25 was used to qualify and quantify the testolactone,respectively.Results Chromatograms showed no endogenous interfering peaks with the urine blank sample.Each analysis was completed within 7min The calibration wag linear in the concentration range within 0.1～50μg/ml.The intra-batch and inter-batch RSD were less than 10%.The recovery rate of the extraction was about 60%.Conclusions The method is proved to meet the requirements of WADA and be suitable for routine screening.

Objective To detect the serum levels of 25‐hydroxy vitamin D[25‐(OH)D] ,homocysteine(HCY) andβ2‐microglob‐ulin(β2‐MG) in the patients with type 2 diabetes mellitus (T2DM ) and to investigate the relationship between serum HCY and β2‐MG with 25‐(OH)D and its clinical significance .Methods A total of 139 cases of T2DM were selected anddivided into 3 groups , the normal albuminuria group for [urinary albumin to creatinine ratio (UACR)< 30 mg/gCr ,45 cases] ,microalbuminuria group (UACR ≥ 30 mg/gCr and < 300 mg/gCr ,48 cases) and massive proteinuria group (UACR ≥ 300 mg/gCr ,46 cases) according to the urinary albumin to creatinine ratio (UACR) .Other 45 individuals undergoing the physical examination were selected as the con‐trol group .The serum 25‐(OH)D level was measured by electrochemiluminescence .Serum HCY level was determined by the enzy‐matic method .Serum β2‐MG level was measured by the latex enhanced immune turbidity method .At the same time ,the biochemical indicators of FBG ,HbA1C ,serum calcium and phosphorus were measured .Results The serum 25‐(OH)D level was decreased with the increase of urinary albumin in the DM patients .And the serum 25‐(OH)D level in the microalbuminuria group and the massive proteinuria group was significantly decreased compared with the normal albuminuria group and the control group ,the difference was statistically significant(P<0 .01) .The serum HCY and β2‐MG levels in the microalbuminuria group and the massive proteinuria group were significantly increased compared with the healthy control group ,the difference was statistically significant (P<0 .05) . Conclusion The serum 25‐(OH)D level is decreased with the increase of urinary albumin in the diabetic patients .The serum HCY andβ2‐MG levels are increased with the increase of urinary albumin and serum 25‐(OH)D level is negatively correlated with the HCY andβ2‐MG levels .

Objective: To discuss the value of femoral head coverage (FHC) in the follow-up of infants with developmental dysplasia of the hip(DDH). Methods: Two hundred and seventeen DDH infants(434 hips) diagnosed by clinical examination and ultrasonography were chosed as treatment group, and then received the brace treatment and followed up with ultrasonography. Four hundred and sixteen infants(832 hips) with normal hips diagnosed by clinical examination and ultrasonography were chosed as control group. The ultrasonographic examination included Graf method and FHC measurement. The infants in the treatment group were followed up with ultrasonography examination up to the age of 6 months. Results: The FHC in the control group was (59.69±4.36)%. In the treatment group, 302 hips were diagnosed to be DDH.Among them, 36 hips were Graf type Ⅱa, 132 hips were Graf type Ⅱb, 69 hips were Graf type Ⅱc, 8 hips were Graf type D, 41 hips were Graf type Ⅲ, and 16 hips were Graf type Ⅳ. The FHC of these hips were (51.33±5.18)%, (53.07±3.94)%, (44.52±5.32)%, (38.70±5.85)%, (27.23±6.82)% and 0, respectively. After treatment, 222 hips became Graf type Ⅰ, 49 hips were improved, and their FHC increased. Twenty-eight hips were not significantly improved after treatment, but the FHC of most hips (including 14 hips of type Ⅱb, 3 hips of Ⅱc and 4 hips of type Ⅲ) increased. Three hips of Graf type Ⅲ aggravated to Graf type Ⅳ, and the FHC decreased to 0. Conclusions The FHC has important value in the follow-up of infants with DDH.

Objective:To measure the pubo-femoral distance of hips of the infants diagnosed with Graf type Ⅰ by high frequency ultrasound.Methods:A total of 1 084 infants(2 168 hips) diagnosed with Graf type Ⅰ by ultrasonography were collected in Beijing Jishuitan Hospital from June 2019 to May 2020. The ultrasonographic examinations included Graf method and pubo-femoral distance measurement. The 95% medical reference value was adopted to confirm range of normal values of the pubo-femoral distance. And the differences of the pubo-femoral distance between different gender, age and bilateral hip were compared.Results:The pubo-femoral distance of hips diagnosed with Graf type Ⅰ was (0.27±0.06)cm, the pubo-femoral distance of males was(0.27±0.06)cm, the pubo-femoral distance of females was (0.27±0.07)cm. The 95% medical reference value was 0.15-0.38 cm. The 95% medical reference value of males was 0.15-0.39 cm, and the 95% medical reference value of females was 0.13-0.41 cm. There were no significant differences between the sexes or between the age(all P>0.05). The difference between the bilateral hip joints from the same infant was statistically significant[(0.274±0.064)cm vs (0.265±0.064)cm, P<0.05]. Conclusions:The 95% medical reference value of hip diagnosed with Graf type Ⅰ is 0.15-0.38 cm.

Objective:To compare the effects of sodium butyrate(Sob)on aged and young asthmatic mice.Methods:Male C57BL/6 mice aged 24 months(n=18)and 4 months(n=18)were divided using randomly generated numbers into 3 groups: a control group, a model group(treated with ovalbumin, OVA), and a treatment group(OVA+ Sob), with 6 mice in each group.The control group was given normal saline as the blank control.Two days before the OVA challenge, the treatment group was given 1 mg/g Sob by intraperitoneal injections and the OVA+ Sob group and the control group were given normal saline intervention.Endpoint evaluation was performed 1 day after the last challenge.Differences in airway hyperresponsiveness(AHR), pathological manifestations of lung tissue sections stained with hematoxylin eosin(HE), and inflammatory factors in lung tissues were compared between aged and young mice and the effects of Sob on asthmatic mice of different ages were evaluated.Results:Under the stimulation of methacholine(Mch), AHR of the O-OVA group was significantly higher than that of the Y-OVA group(6.250 g/L: 276.28±113.62 vs.103.02±19.55, t=3.368, P=0.026; 12.500 g/L: 457.5±157.29 vs.114.76±20.28, t=4.338, P=0.022; 25.000 g/L: 1113.16±256.98 vs.567.87±187.34, t=3.538, P=0.009). HE staining revealed that, compared with the Y-control group, the O-control group exhibited a reduced area of alveolar cavity, partial lung consolidation, proliferation of interstitial fibrous connective tissues, alveolar epithelial cells and capillary endothelial cells.Compared with its control group, the O-OVA group had significantly elevated levels of eotaxin( P=0.035), IL-17A( P=0.004)and IL-1β( P=0.001), among the inflammatory factors, whereas the Y-OVA group had elevated levels of eotaxin( P=0.001), IL-17A( P=0.001), IL-4( P=0.004), KC( P=0.012)and IL-1β( P<0.001). Compared with the O-OVA group, the Y-OVA group had increased levels of IL-4( Z=2.882, P=0.004)but decreased levels of IL-1β( t=2.728, P=0.020). As for the effects of Sob on asthmatic mice of different ages, AHR of the O-OVA+ Sob group was significantly alleviated with stimulation of 12.500 g/L Mch( P=0.015)and 25 g/L Mch( P=0.014), compared with the O-OVA group.With stimulation of 3.125 g/L Mch, AHR of the Y-OVA+ Sob group was significantly decreased( P=0.021)and levels of IL-4( P=0.004)and IL-1β( P=0.014)were significantly reduced in the Y-OVA+ Sob group, compared with the Y-OVA group. Conclusions:The severity of asthma in aged mice is greater than in young mice, perhaps as a result of different immunophenotypes; The alleviating effects of Sob on inflammatory factors in young asthmatic mice may be related to mild AHR in young asthmatic mice, compared with aged asthmatic mice.

Objective? To investigate the health belief of colorectal cancer and its screening in relatives of patients with hereditary colorectal cancer,and to analyze its influencing factors. Methods? Totally 148 blood relatives of patients with hereditary colorectal cancer hospitalized in the Department of Colorectum of a Class ⅢGrade A hospital in Guangzhou from December 2016 to May 2018 were selected using convenient sampling, and investigated with the self-made general information questionnaire and Chinese Colorectal Cancer Health Belief Scale (CCCHBS). SPSS 20.0 was used for statistical analysis. Results? The colorectal cancer health belief of blood relatives of patients with hereditary colorectal cancer totaled (3.37±0.35) in which the perception and acceptance levels of screening benefits, health motives and self-efficacy of colorectal cancer were higher, while the perception levels of severity of colorectal cancer, barriers against colorectal screening and susceptibility were lower. Multivariate linear regression analysis revealed that household income per capita, educational background, with commercial medical insurance or not and the number of blood relatives with cancer were the influencing factors for health belief levels, which accounted for 41.9% of the total variance. Conclusions? The health belief in blood relatives of patients with hereditary colorectal cancer remain at a medium level. Medical and nursing workers should pay attention to the blood relatives of patients with hereditary colorectal cancer with lower educational background, lower household income and less blood relatives with cancer, take measures to improve their health belief levels, and facilitate colorectal cancer screening behaviors so as to prevent and control colorectal cancer in a better manner.

Objective:To explore the high-frequency ultrasonographic and clinical features of superficial intravascular tumors.Methods:Thirteen superficial intravascular tumor patients who underwent ultrasound examination form Beijing Jishuitan Hospital during 2016-2020 were retrospectively analyzed. The location, anatomic blood vessels, shape, size, boundary, internal echo, blood flow and clinical characteristics of the tumors were analyzed.Results:Among the 13 cases, most of the tumors were oval like or long strip, and a few were round like. All 11 cases of primary intravascular tumors showed hypoechoic mass; 2 cases of intravascular metastasis or recurrence showed solid hypoechoic with hyperechoic mass. Arterial blood flow spectrum was detected in 12 cases. Two patients with glomus tumor showed typical tenderness at the fixed point. Two patients had a history of bone tumors in the upper or lower extremities.Conclusions:High frequency ultrasound can clearly recognize the location of the superficial intravascular tumors, describe the morphological characteristics and sonographic features of the tumor, then help to make a more accurate qualitative diagnosis.

Objective:To analyze the genetic and clinical characteristics of MYO15A variants associated non-syndromic autosomal recessive deafness3 (DFNB3).Methods:The hearing test and high-throughput sequencing data of 108 families with non-syndromic hearing loss, who visited the Center of Genetics and Prenatal Diagnosis in the First Affiliated Hospital of Zhengzhou University from November 2016 to February 2019, were retrospectively analyzed to investigate the characteristics of MYO15A variation.Results:Compound heterozygous MYO15A variations were detected in nine patients from eight families, accounting for 7.4% of all 108 families. The variants were c.5910+1G>A/c.9417_9418insTA, c.4234T>G/c.8324G>T, c.3926A>T/c.5002delC, c.9690+1G>A/c.10257_10259delCTT, c.8324G>T/c.10419_10423delCAGCT, c.4519C>T/c.6454G>C, c.6177+1G>T/c.10257_10259delCTT and c.5692C>T/c.7396-1G>A. All patients had severe to profound hearing loss. Among the 14 variations, 12 variations were located in the main structural domains, including 5 in motor domain, 3 in FERM domain, 3 in MyTH4 domain and 1 in IQ motif. The c.3926A>T, c.4234T>G, c.4519C>T, c.5002delC, c.6454G>C, c.8324G>T, c.9417_9418insTA and c.10419_10423delCAGCT had not been reported in the Human Gene Mutation Database up to February 2020. According to the guidelines of the American College of Medical Genetics and Genomics (ACMG), 6 reported variants and the first reported c.4519C>T, c.5002delC, c.9417_9418insTA and c.10419_10423delCAGCT were identified as pathogenic variants, while c.8324G>T was likely pathogenic variant, and c.3926A>T, c.4234T>G and c.6454G>C were variants of uncertain significance.Conclusions:The variations of MYO15A in patients with DFNB3 are mainly complex heterozygous. The clinical phenotypes are mostly severe to profound hearing loss, and the mutation loci are mainly in the motor, FERM and MyTH4 domains.

Objective:To discuss the value of pubo-femoral distance(PFD) in the diagnosis of infants with developmental dysplasia of the hip(DDH).Methods:Nine hundred and thirty-one infants with DDH and normal infants diagnosed by clinical examination and ultrasonography were involved from February 2022 to June 2023 in Beijing Jishuitan Hospital. The ultrasonographic examination included Graf method and pubo-femoral distance measurement. Statistical analysis was made on the measurements of PFD of different types of hips.Results:Among all the hips, 1 359 hips were Graf type Ⅰ, 274 hips were Graf type Ⅱa, 166 hips were Graf type Ⅱb, 31 hips were Graf type Ⅱc, 7 hips were Graf type D, 13 hips were Graf type Ⅲ, and 12 hips were Graf type Ⅳ. The PFD of these hips were (0.34±0.07)cm, (0.35±0.08)cm, (0.35±0.08)cm, (0.51±0.11)cm, (0.67±0.09)cm, (0.95±0.26)cm, (1.27±0.30)cm, respectively. There was no significant difference in PFD between hips of Graf Ⅰ, Graf Ⅱa and Graf Ⅱb ( P>0.05), but there was significant difference in PFD between hips of the other Graf types ( P<0.05). The area under curve(AUC) of the PFD for the diagnosis of stability of the hips in infants was 0.959 with the cut-off value of 0.435 cm, the sensitivity and specificity were 88.9% and 90.4%, respectively. Conclusions:The PFD has great value in the evaluation of the stability of hip joint, and the cut-off value is 0.435 cm.

Rabdosia serra(R.serra),an important component of Chinese herbal tea,has traditionally been used to treat hepatitis,jaundice,cholecystitis,and colitis.However,the chemical composition of R.serra and its effect against colitis remain unclear.In this study,the chemical composition of the water extract of R.serra was analyzed using ultra performance liquid chromatography coupled with a hybrid linear ion trap quadrupole-orbitrap mass spectrometer(UPLC-LTQ-Orbitrap-MS).A total of 46 compounds,comprising ent-kaurane diterpenoids,flavonoids,phenolic acids,and steroids,were identified in the water extract of R.serra,and the extract could significantly alleviate dextran sulfate sodium salt-induced colitis by improving colon length,upregulating anti-inflammatory factors,downregulating proinflammatory fac-tors,and restoring the balance of T helper 17/T regulatory cells.R.serra also preserved intestinal barrier function by increasing the level of tight junction proteins(zonula occludens 1 and occludin)in mouse colonic tissue.In addition,R.serra modulated the gut microbiota composition by increasing bacterial richness and diversity,increasing the abundance of beneficial bacteria(Muribaculaceae,Bacteroides,Lactobacillus,and Prevotellaceae_UCG-O01),and decreasing the abundance of pathogenic bacteria(Turi-cibacter,Eubacterium_fissicatena_group,and Eubacterium_xylanophilum_group).Gut microbiota depletion by antibiotics further confirmed that R.serra alleviated colitis in a microbiota-dependent manner.Overall,our findings provide chemical and biological evidence for the potential application of R.serra in the management of colitis.