In this study ,the objective is to establish a nested‐PCR assay for the detection of H .bilis with high sensitivity and specificity .The nested primers were designed based on sequences of 16S rRNA gene of seventeen subtypes of H .bilis .Af‐ter optimizing reaction condition ,the sensitivity and specificity of the assay were examined via the detection of feces simulated samples ,mice infection model samples and clinic patients’ samples .The detection sensitivity of H .bilis strain for feces simu‐lated samples was 10 CFU/100 μL .H .bilis was successfully detected in the liver ,caecum and feces of experimentally infected mice .Moreover ,H .bilis was successfully detected in the bile ,cholecyst mucous membrane and feces samples from two of ten patients with cholelithiasis .Due to the PCR assay’s high sensitivity and specificity ,the method may be used to detect the infec‐tion of H .bilis .

Objective To evaluate the effect of exogeneous adiponectin on hippocampal advanced glycation end products (AGEs)-reactive oxygen species (ROS)-endoplasmic reticulum stress (ERS) pathway in aged mice with postoperative cognitive dysfunction (POCD).Methods Thirty-two healthy male C57BL/6 mice, aged 18 months, weighing 20-25 g, were randomly divided into 4 groups (n=8 each) using a random number table: control group (group C), POCD group, exogeneous adiponectin group (group APN), and vehicle group (group Veh).Splenectomy was performed to establish the POCD model in aged mice anesthetized with intraperitoneal pentobarbital sodium.In group APN, adiponectin 0.1 μg/g (in 2 μl of phosphate buffer solution) was injected into the lateral cerebral ventricle at 30 min before establishing the model.Phosphate buffer solution 2 μl was given at 30 min before establishing the model in group Veh.Cognitive function was assessed on day 7 after surgery.The mice were then sacrificed, and the hippocampus was harvested for determination of the area of AGE deposition (by immunohistochemistry), levels of ROS (by flow cytometry), and levels of glucose-regulated protein 78 (GRP78), C/EBP-homologous protein (CHOP), caspase-12 and ROS (using Western blot).Results Compared with group S, the freezing time in the contextual fear conditioning test was significantly shortened, the area of AGE deposition and levels of ROS, CHOP and caspase-12 were increased, and the level of GRP78 was decreased in POCD, APN and Veh groups.Compared with POCD and Veh groups, the freezing time in the contextual fear conditioning test was significantly prolonged, the area of AGE deposition and levels of ROS, CHOP and caspase-12 were decreased, and the level of GRP78 was increased in group APN.Conclusion Exogeneous adiponectin decreases the occurrence of POCD probably by blocking hippocampal AGEs-ROS-ERS pathway in aged mice.

Objective:To explore the relationship between DNA ploidy heterogeneity and clinical biological behavior on patients with malignant tumor.Methods:The DNA ploidy heterogeneity of tumor tissue was measured in 163 patients with malignant tumors by flow cytometry.The relations were analyzed between DNA ploidy heterogeneity and clinical stage,pathological grade,metastasis rate of patients with malignant tumors.Results:The rates of DNA ploidy heterogeneity were significantly different in different tumors.The rates of heterogeneity raised with increase of clinical stage and pathological grade(P

Objective:To investigate a large Chinese family in which 9 patients over 4 generations were diagnosed with a form of autosomal dominant spondyloepimphyseal dysplasia(SEMD).Mothods:X-Ray radiograph of proand at 18-month showed absence of secondary ossification centra of femoral heads.His father at 24-year presented severe spondyloepiphyseal changes that principally involved the vertebral bodies,the femoral necks and femoral heads and characterized by generalized platyspondyly with thoracolumbar scoliosis,irregular femoral necks,absent ossification of femoral heads,flat acetabular roofs and coxa vara.The other patients had similar clinical and radiological features.Haplotyping was performed with leukocyte DNA for 5 micosatellite repeat markers from chromosome 12 and the result showed COL2A1 gene as a candidate gene.A total of 54 exons and promoter of COL2A1 gene were amplified and sequenced from all patients and available normal relatives.In addition,exon 23 of COL2A1 gene was amplified and sequenced from 10 controls simultaneously.Results:All patients were identified a 1510(G→A) transition in exon 23 of COL2A1 gene that caused a change from a COL2A1 coding region in available glycine to serine at amino acid position 504.No mutation was found in the normal relatives and 10 controls. Conclusion:The mutation of COL2A1 gene is responsible for this form of SEDC of the family.This is the first familial report of SEDC relating to 1510G→A mutation of COL2A1 gene.The detailed clinical radiogram data will be useful for extending the phenotypic spectrum of type Ⅱcollagenopathies.

Objective: To report a case of azoospermia with a karyotype of 45,X,der(Y)t(Y;13)(q11.2;q12),-13,accompanied with slight bilateral gynecomastia and multiple nodules.Methods: The karyotype was identified by karyotyping and FISH,and the breakpoints of the Y chromosome and the copy number of the BRCA2 gene in 13q12 determined by PCR-STS and DNA polymorphic analysis.The testis and nodule tissues of the patient were obtained for biopsy.Results: FISH confirmed SRY and centromere of the Y chromosome on the questionable 13 chromosome and the karyotype to be 45,X,der(Y)t(Y;13)(q11.1;q12),-13.ish der(Y)(SRY+,DYZ3+,wcp13+).PCR-STS showed the deletion of regions AZFa,b and C,with a breakpoint located inYq11.1 below sY82.No deletion of the BRCA2 gene was observed.The patient was diagnosed with Sertoli cell-only syndrome by testicular biopsy and with angiolipomata by pathological examination of the nodule tissue.Conclusion: The patient's phenotype of complete masculinization could be attributed to presence of the SRY gene,and his azoospermia with small testis to the absence of a fragment from Yq11.1 to Yqter.However,the molecular mechanism of angiolipoma remains unknown.

Objective To report the prenatal molecular diagnosis for two gravida in a family with spondylepiphyseal dysplasia congenita(SEDC)caused by G504S mutation of COL2A1 gene.Methods DNA of the two fetuses was extracted from amniotic fluid at the 19+3 and 18+6 weeks of gestation respectively.Direct sequencing of two samples were performed after amplifying exon 23 of COL2A1 containing the potential mutation.The femur length and biparietal diameter of the first fetus were measured by sonographic scans every two weeks from 17+3 weeks to 27+3 weeks of gestation,and for the second fetus these parameters were measured from 16+1 to 19+1 weeks of gestation.Results Sequncing analysis revealed the first fetus and his mother presented the same mutation which is specifically associated with SEDC,but the second fetus did not show the mutation of COL2A1 gene.Biparietal diameters of the both fetuses were appropriate for gestational age.Femur length of the second fetus was normal for gestational age but that of the first fetus was shortened evidently after the 23 week of gestation.The parents of the first fetus determined to terminate the pregnancy.A medical termination was carried out at 27+5 weeks of gestation and a male fetus with a relatively large head and short limbs was delivered.The radiological findings of the fetus were consistent with SEDC including generalized platy spondesand shortened long bones.Conclusions Prenatal molecular diagnosis is important for the fetus with risk of SEDC and useful for genetic counseling.Genotype of fetus with risk of SEDC can be identified before sonographic scan.Molecular genetic analysis in conjunction with sonographic monitoring was helpful in prenatal diagnosis of SEDC.

Objective:To explore the clinical effects in repairing dorsal complex tissue defect of finger with the flap based on superficial palmar branch of radial artery (SPBRA) with palmaris longus tendon.Methods:From May, 2011 to October, 2017, dorsal complex tissue defects of 15 fingers and thumbs in 15 patients were treated by the flaps which were based on SPBRA with palmaris longus tendon. There were 10 males and 5 females, in an average of 35.8 (19-51) years. All the defects (3 thumbs, 5 index, 2 middle and 5 ring fingers) were dorsal complex tissue defects and all had extensor tendon defects. The areas of soft tissue defect measured 2.0 cm×1.5 cm to 4.0 cm×2.0 cm. The lengths of tendon defect measured 2.0-4.0 cm. All patients received emergency surgery. The time before the surgery was 1.5-3.0 hours. The surgery time was 3.0-4.5 hours, 3.6 hours in an average. Postoperative regular follow-up.Results:All of the wounds healed in stage I and all flaps survived. Texture of the flaps was soft with rosy color. No obvious swelling occurred. All the donor sites healed in stage I. The patients were followed-up for 4-18 months, 8 months in an average. The appearance and function of the repaired fingers and thumbs were satisfactory. The pain, temperature and touch sensations were good. The color of flaps was similar to the normal finger without swelling. The wear resistance of the flaps was good. Thin-line scars were in the wrist donor sites without contracture. The range of motion of active palmar flexion of the wrist was from 0° to 80° and active hyperextension was from 0° to 70°. No obvious limitation was found.Conclusion:It is able to achieve a satisfactory clinical effects by using the flap that is based on SPBRA with palmaris longus tendon in repairing the dorsal complex tissue defect of hand. The advantages of the technique are that the donor site is concealed. The wound is small, and the flap is easy to be harvested and anastomosed.

Objective:To predict and analyze the number of acute pancreatitis (AP) inpatients based on time series model, and to explore the predictive efficiency of the model.Methods:Clinical data of AP inpatients in the Affiliated Hospital of Southwest Medical University from January 2014 to December 2019 were collected. R software was used to collect the time series of AP inpatients, and the trend and seasonal characteristics of AP inpatients from 2014 to 2018 were analyzed. Furthermore, the autoregressive moving average (ARIMA) model was established through stationarity test, model ordering and model testing steps, and the best selected model was used to predict the monthly number of inpatients in 2019 to verify its prediction efficiency.Results:A total of 3 939 AP patients were included in the study. The most common etiology for AP was cholestrogenic (48.2%), followed by hyperacylglyceremia (36.3%). The peak age of hospitalization was from 40 to 60 years old. Time series analysis showed that the number of AP inpatients increased year by year. The highest peak of the disease was from February to March, followed by September to November; and there was seasonal variation and the incidence was relatively small in summer. The established original training set sequence did not pass the stationarity test ( P=0.061), so the ARIMA model was established after it was transformed into a stationarity sequence by first-order difference. According to the criterion of minimum AIC value, ARIMA(2, 1, 1)(1, 1, 1) 12 was selected as the best model. The model was used to predict the number of AP inpatients in 2019, showing that it could better fit the trend of onset time and had good short-term prediction effect. The mean root error and absolute error were 6.8790 and 4.7783, respectively. Conclusions:The number of AP inpatients increases year by year with seasonal changes. ARIMA model is effective in predicting the number of AP inpatients and can be used for short-term prediction.

Objective:To access the efficacy of monoenergetic imaging from spectral CT combined with metal artifact reduction for orthopedic implants (O-MAR) on reducing contrast hardening artifacts in the vein on the injection side, and determining the optimal monoenergetic spectral range to improve the display of axillary lymph node.Methods:A total of 35 patients with breast cancer who underwent chest-enhanced CT scans were enrolled in this retrospective study. The original data were reconstructed to obtain a total of 35 sets of images, including one conventional image, 17 groups of monoenergetic images, and 17 groups of monoenergetic+O-MAR images. The areas of interest were delineated in the high and low-density artifact area on the injection side of the same layer contrast agent, and the contralateral ectopectoralis. The CT value and its standard deviation (SD) were recorded respectively, the artifact area was measured, and the number of axillary lymph nodes was recorded. The difference in CT values (ΔCT 1, ΔCT 2) and the artifact index (AI1 and AI 2) of the high and low-density artifact areas relative to the contralateral ectopectoralis in the same layer were calculated respectively. Friedman test and Wilcoxon signed-rank test were used to compare the differences of ΔCT, AI, artifact area, and number of lymph nodes among the three imaging modalities, and the Kappa test was used to compare the differences in subjective evaluation. Results:As the energy level increased, compared to the conventional image, monoenergetic image, ΔCT 1 absolute value, ΔCT 2 absolute value, AI 1, and AI 2 showed a trend of initially low and then high, artifact area decreased, and the number of detected lymph nodes increased ( P<0.01). Compared to other energy levels, when the monoenergetic image was 100 keV, ΔCT 1 value, 140 keV for ΔCT 2 value, 120 keV for AI 1 value, and 130 keV for AI 2 value were close to zero, and the number of detected lymph nodes was highest at 110-200 keV. In contrast, in the monoenergetic+O-MAR images, ΔCT 1 absolute value showed a trend of initially low and then high, but, ΔCT 2 absolute value, AI 1, AI 2, and artifact area all significantly decreased, whereas the number of detected lymph nodes significantly increased (χ 2 values were 916.23, 895.93, 387.08, 519.41, 890.10, and 1027.98, respectively. All P<0.01). Compared to other energy levels, when the monoenergetic+O-MAR image was at 100 keV, ΔCT 1 value was close to zero, while ΔCT 2 value became close to zero with increasing energy level, and the number of detected lymph nodes was highest at 110-200 keV. As the energy level increased, the ΔCT 1, AI 1, AI 2, and artifact area of monoenergetic+O-MAR images were significantly smaller than those of monoenergetic images at the same energy level, and the number of detected lymph nodes was significantly higher than that of monoenergetic images ( P<0.01). The subjective scores for 110-200 keV monoenergetic images and 100-200 keV monoenergetic+O-MAR images were both higher than 4, and the score for monoenergetic+O-MAR images was significantly higher than that of single-energy spectrum images. The agreement between the two radiologists in assessing subjective scores was good. Conclusion:At 100-120 keV level, spectral CT monoenergetic combined with O-MAR imaging technique has the best performance in removing hardening-induced artifacts of chest-enhanced CT contrast agent and detecting and displaying axillary lymph nodes.

Objective To analyze the clinical characteristics and genotype of a patient with congenital systemic lip-odystrophy(CGL)type 1 associated with exudative xanthoma caused by AGPAT2 gene mutation,and to provide ev-idence for clinical and genetic diagnosis of the disease.Methods Clinical data of the patient such as medical histo-ry,physical examination and laboratory examination were collected.Peripheral venous blood was collected for whole exome sequencing analysis and Sanger sequencing verification,and treatment was provided to patients according to the changes of condition.Results The clinical manifestations of the patient were subcutaneous fat reduction,fatty liver,spleen enlargement,kidney enlargement,high blood sugar and lipids,severe insulin resistance,scattered yellow rash on limbs,which was confirmed as xanthoma.The results of whole exon sequencing showed that the AGPAT2 gene of the patient had a heterozygous nonsense mutation of c.202C＞T:p.R68?and c.646A＞T:p.K216?,and the former was the pathogenic mutation site.Follow-up therapy covers improvement of lifestyle,low-fat diet and regular exercise.The rashes subsided after active lipid-lowering therapy.Conclusions Apart from typical lipody-strophy,the patient was accompanied by exanthemous xanthoma.No CGL1 patient with exanthemous xanthoma has been reported in the domestic literature database up to now,and the genetic test results showed that there was a c.202C＞T heterozygous mutation of AGPAT2 gene.This gene site has not been reported in the literature,and its functional verification needs to be further studied.