Objective To explore the related factors of the influence of persistent pulmonary hypertension( PPH) after neonatal resuscitation. Methods Tow hundred and fifty?six cases asphyxia patients who were treated in the Maternal and Child Care Service Center of Shangluo from March 2012 to March 2015 were retrospectively analyzed. One hundred and thirty?six newborns asphyxia of persistent pulmonary hypertension were selected as the observation group. One hundred and twenty newborns asphyxia without persistent pulmonary hypertension were selected as the control group. Medical records of children of two groups were retrospectively analyzed. Apgar scores of all observed objects. Before resuscitation blood pressure, blood glucose, body temperature,blood pH value of children in the two groups were recorded. Results In admission, the systolic blood pressure,diastolic blood pressure,body temperature,blood sugar,blood pH value in the observation group ((49. 27±8. 08) mmHg,(22. 25±5. 17) mmHg,(34. 40±0. 41)℃,(2. 78±0. 65) mmol/L,(7. 11±0. 79)) were lower than that in the control group((51. 23±4. 17) mmHg,(23. 87±2. 18) mmHg,(36. 80±0. 85)℃, (3. 55±0. 33) mmol/L,(7. 33±0. 17)),the differences were statistically significant(t=2. 480,3. 334,28. 169, 12. 155,3. 166;P<0. 05) . Hypoglycemia,hypothermia,low systolic blood pressure,low diastolic blood pressure were identified as the impact factors that can cause neonatal resuscitation after persistent pulmonary hypertension ( OR=3. 660,1. 930,8. 903,4. 543;95%CI=1. 061-12. 631,1. 399-2. 661,1. 166-67. 962,1. 636-12. 613;P<0. 05). Conclusion Hypoglycemia,hypothermia,low systolic blood pressure and low diastolic blood pressure are impact factors can cause neonatal resuscitation after persistent pulmonary hypertension.

Objective To prokaryotically express the valosin-containing protein(VCP)of Schistosoma japonicum,and ana-lyze its VCP mRNA expressions in the cercaria,schistosomulum,adult worm(female and male worms)and egg. Methods RNA of S. japonicum eggs were extracted,and reversely transcribed into cDNA. The VCP gene of S. japonicum was amplified by using polymerase chain reaction(PCR),and subcloned into the prokaryotically expressed vector pET15b. The recombined plasmid was transformed into BL21 cells,and the expression of the target gene was induced with isopropyl-beta-D-thiogalactopyranoside (IPTG). The recombinant protein was yielded through the purification of inclusion body,and identified by using sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE). The RNA(s)of cercaria,schistosomulum,female adult worm,male adult worm,and egg of S. japonicum were extracted,digested with DNase,purified,and reversely transcribed into cDNA. The mRNA expressions of the VCP gene in various developmental stages of S. japonicum were determined by using fluorescence-based quantitative real-time PCR. Results The VCP gene of S. japonicum was yielded by PCR amplification,and the recombinant pro-tein was obtained through recombinant plasmid expression and purification of inclusion body. The highest VCP mRNA expression in S. japonicum cercaria was detected by the fluorescence-based quantitative real-time PCR,while low expressions were found in the schistosomulum,egg,female and male adult worms. Conclusion The recombinant protein encoded by the VCP gene of S. ja-ponicum is successfully obtained,and the VCP mRNA expression is determined in various developmental stages of S. japonicum.

Objective:To investigate the clinical effect of botulinumtoxin type A (Botox-A) combined with electromyographic biofeedback therapy on the upper limb muscle spasm after stroke.Methods:86 cases of patients with upper limb muscle spasm after stroke in our hospital from January 2016 to January 2017 were selected and divided into the observation group and the control group,with 43 cases in each group.Patients in the control group were treated with electromyographic biofeedback therapy,and the observation group was treated with Botox-A based on the basis of control group.The improvement of upper limb muscle spasm,Upper limb movement function,the active range of wrist joint and life skills before and after treatment were compared between two groups.Results:After treatment,the total effective rate of improvement of upper limb muscle spasm of observation group were significantly higher than that of the control group (P＜0.05);At 2 weeks and 4 weeks after treatment,the Fugl-Meyer scores,Wrist joint activities,modified Barthel index (MBI) of two groups were significantly higher than those before treatment (P＜0.05),which were significantly higher in the observation group than those of the control group (P＜0.05).Conclusion:Botox-Acombined with electromyographic biofeedback therapy had remarkable clinical effect on the upper limb muscle spasm after stroke,which could effectively reduce the upper limb spasticity,improve the arm and wrist movement ability and the ability of daily life.

BACKGROUND: It has been suggested that amniotic epithelial cells (AECs) express almost all the markers of neural cell and secrete biologically active neurotrophins such as brain derived neurotrophin factor (BDNF) and neurotrophin-3 (NT3).If AECs can substitute neural cells, its neurotrophic effect will bring expansive prospect in treating spinal cord injuries and degenerative neural disease.OBJECTIVE: To observe the survival, migration and secretory function of AECs after transplanted into the injured spinal cord.DESIGN: An observational experiment.SETTING: Department of Histology and Embryology, School of Basic Medical Science, Jilin University.MATERIALS: Embryonic rat of 12-14 days (n =1) and adult Wistar rats (n =18, 300-350 g) were provided by the Experimental Animal Center of Jilin University. Immunohistochemical reagents: Mouse anti-rat BrdU monoclonal antibody was bought from Sigma Company. Rabbit anti-rat NT3 polyclonal antibody and rabbit anti-rat BDNF polyclonal antibody were bought from Boster Company. SP immunohistochemistry reagents were purchased from Maixin Company.METHODS: The experiment was made in the Department of Histology and Embryology, Basic Medical Science of Jilin University from July to October 2005. ① Wistar rats were anesthetized by intraperitoneal injection of chloral hydrate, subcutaneous tissue and muscle were separated, spinous process and lamina of vertebra were removed by bone ribbing rongeur. to expose the spinal cord. The spinal cords were clamped at the twelfth thoracic vertebra (T12) for 3 minutes.After surgery, the wounds were smeared with penicillin G, then muscle and skin were sutured. The rats were anesthetized by inhaling ether if necessary. ② Obtaining and culture of AECs: Amniotic membrane was peeled from the placenta of a pregnant Wistar rat of 12-14 days. The amnictic membrane was dissected into small pieces of 1 mm×1 mm×1 mm, then digested and cultured, and mechanically made into single cell suspension, finally plated in bottles. ③ Transplantation of AECs into injured spinal cord: The initial wound was slit and injected with 5 μL Brdu labeled AECs (1×1012 L-1) to the exposed injured spinal cord at 3.0 mm anterior to the injured site. The injections were made at a rate of 5 μL per 3 minutes with a microsyringe. The syringe was slowly pulled out after 5 minutes, then muscle and skin were sutured. ④ Sampling and immunohistochemical analysis: Three animals were sacrificed at 1 week and the other three at 2 weeks postoperatively. The sections were fixed with 40 g/L paraformaldehyde in phosphate buffer solution (PBS) for 20 minutes at room temperature, followed by incubation with primary antibodies at 4 ℃ overnight. The samples were treated with secondary antibodies, biotinylated anti-mouse or rabbit immunoglobulin (IgG) at 37 ℃ for 20 minutes; Followed by incubation of horseradish peroxidase (HRP) labeled third antibodies at 37 ℃ for 20 minutes, then stained with 0.2 g/L diaminobenzidine (DAB) or AEC.MAIN OUTCOME MEASURES: Survival, migration and expression of AECs after transplanted into the injured spinal cord. RESULTS: After transplantation, most of the AECs gather beneath the pia mater of injured spinal cord at 1 week. But they migrated more extensively and many positive nuclear cells (brown) were observed in the center cannel and surrounding gray mater. Meantime, it was also detected that the transplanted AECs could express NT3 (positive cells stained as red) and BDNF in the injured spinal cord.CONCLUSION: AECs could survive for at least 3W after transplanted into the injured spinal cord of adult rats and could migrate widely; Furthermore, they could secrete neurotrophic factors such as NT-3 and BDNF.

Objective To identify the diagnostic value of ADC combined with DWI in benign lesions and malignant lesions of testis. Methods 35 patients with testicular lesions confirmed by operation and pathological examination in our hospital were analyzed retrospectively, including 18 benign lesions and 17 malignant lesions.The mean ADC values of normal tissue and parenchyma of testicular lesions were measured and statistically analyzed by K ruskal-W allis test,and receiver operating characteristic(ROC)curve was delineated. The optimum ADC value for differential diagnosis of malignant testicular lesions was analyzed and determined.Results In 33 cases of normal testicular tissue DWI showed homogeneous high signal,and mean ADC was(1.137 ± 0.119)×10-3mm2/s.18 cases of benign lesions mostly showed unrestricted diffusion,and mean ADC was(1.104 ± 0.463)×10-3mm2/s.In 17 cases of malignant lesions DWI showed high signal,and mean ADC was(0.778 ± 0.198)×10-3mm2/s.The comparison of ADC mean values between malignant testicular lesions and normal tissue as well as benign lesions of testis showed significant difference(P<0.05).The optimum ADC to distinguish malignant testicular lesions from benign testicular lesions was 0.911×10 -3mm2/s(82.4% sensitivity and 82.4% specificity). Conclusion DWI combined with ADC value is beneficial to the preoperative diagnosis and differential diagnosis between malignant testicular lesions and benign lesions of testis.

Plasmids remain important microbial components mediating the horizontal gene transfer (HGT) and dissemination of antimicrobial resistance. To systematically explore the relationship between mobile genetic elements (MGEs) and antimicrobial resistance genes (ARGs), a novel strat-egy using single-molecule real-time (SMRT) sequencing was developed. This approach was applied to pooled conjugative plasmids from clinically isolated multidrug-resistant (MDR) Klebsiella pneu-moniae from a tertiary referral hospital over a 9-month period. The conjugative plasmid pool was obtained from transconjugants that acquired antimicrobial resistance after plasmid conjugation with 53 clinical isolates. The plasmid pool was then subjected to SMRT sequencing, and 82 assem-bled plasmid fragments were obtained. In total, 124 ARGs (responsible for resistance toβ-lactam, fluoroquinolone, and aminoglycoside, among others) and 317 MGEs [including transposons (Tns), insertion sequences (ISs), and integrons] were derived from these fragments. Most of these ARGs were linked to MGEs, allowing for the establishment of a relationship network between MGEs and/or ARGs that can be used to describe the dissemination of resistance by mobile elements. Key elements involved in resistance transposition were identified, including IS26, Tn3, IS903B, ISEcp1, and ISKpn19. As the most predominant IS in the network, a typical IS26-mediated multi-copy composite transposition event was illustrated by tracing its flanking 8-bp target site duplica-tions (TSDs). The landscape of the pooled plasmid sequences highlights the diversity and complex-ity of the relationship between MGEs and ARGs, underpinning the clinical value of dominant HGT profiles.

Objective:To investigate the diagnostic accuracy of serological indicators and evaluate the diagnostic value of a new established combined serological model on identifying the minimal hepatic encephalopathy (MHE) in patients with compensated cirrhosis.Methods:This prospective multicenter study enrolled 263 compensated cirrhotic patients from 23 hospitals in 15 provinces, autonomous regions and municipalities of China between October 2021 and August 2022. Clinical data and laboratory test results were collected, and the model for end-stage liver disease (MELD) score was calculated. Ammonia level was corrected to the upper limit of normal (AMM-ULN) by the baseline blood ammonia measurements/upper limit of the normal reference value. MHE was diagnosed by combined abnormal number connection test-A and abnormal digit symbol test as suggested by Guidelines on the management of hepatic encephalopathy in cirrhosis. The patients were randomly divided (7∶3) into training set ( n=185) and validation set ( n=78) based on caret package of R language. Logistic regression was used to establish a combined model of MHE diagnosis. The diagnostic performance was evaluated by the area under the curve (AUC) of receiver operating characteristic curve, Hosmer-Lemeshow test and calibration curve. The internal verification was carried out by the Bootstrap method ( n=200). AUC comparisons were achieved using the Delong test. Results:In the training set, prevalence of MHE was 37.8% (70/185). There were statistically significant differences in AMM-ULN, albumin, platelet, alkaline phosphatase, international normalized ratio, MELD score and education between non-MHE group and MHE group (all P<0.05). Multivariate Logistic regression analysis showed that AMM-ULN [odds ratio ( OR)=1.78, 95% confidence interval ( CI) 1.05-3.14, P=0.038] and MELD score ( OR=1.11, 95% CI 1.04-1.20, P=0.002) were independent risk factors for MHE, and the AUC for predicting MHE were 0.663, 0.625, respectively. Compared with the use of blood AMM-ULN and MELD score alone, the AUC of the combined model of AMM-ULN, MELD score and education exhibited better predictive performance in determining the presence of MHE was 0.755, the specificity and sensitivity was 85.2% and 55.7%, respectively. Hosmer-Lemeshow test and calibration curve showed that the model had good calibration ( P=0.733). The AUC for internal validation of the combined model for diagnosing MHE was 0.752. In the validation set, the AUC of the combined model for diagnosing MHE was 0.794, and Hosmer-Lemeshow test showed good calibration ( P=0.841). Conclusion:Use of the combined model including AMM-ULN, MELD score and education could improve the predictive efficiency of MHE among patients with compensated cirrhosis.

Objective:To compare the differences in the prevalence of mild micro-hepatic encephalopathy (MHE) among patients with cirrhosis by using the psychometric hepatic encephalopathy score (PHES) and the Stroop smartphone application (Encephal App) test.Methods:This prospective, multi-center, real-world study was initiated by the National Clinical Medical Research Center for Infectious Diseases and the Portal Hypertension Alliance and registered with International ClinicalTrials.gov (NCT05140837). 354 cases of cirrhosis were enrolled in 19 hospitals across the country. PHES (including digital connection tests A and B, digital symbol tests, trajectory drawing tests, and serial management tests) and the Stroop test were conducted in all of them. PHES was differentiated using standard diagnostic criteria established by the two studies in China and South Korea. The Stroop test was evaluated based on the criteria of the research and development team. The impact of different diagnostic standards or methods on the incidence of MHE in patients with cirrhosis was analyzed. Data between groups were differentiated using the t-test, Mann-Whitney U test, and χ2 test. A kappa test was used to compare the consistency between groups. Results:After PHES, the prevalence of MHE among 354 cases of cirrhosis was 78.53% and 15.25%, respectively, based on Chinese research standards and Korean research normal value standards. However, the prevalence of MHE was 56.78% based on the Stroop test, and the differences in pairwise comparisons among the three groups were statistically significant (kappa = -0.064, P < 0.001). Stratified analysis revealed that the MHE prevalence in three groups of patients with Child-Pugh classes A, B, and C was 74.14%, 83.33%, and 88.24%, respectively, according to the normal value standards of Chinese researchers, while the MHE prevalence rates in three groups of patients with Child-Pugh classes A, B, and C were 8.29%, 23.53%, and 38.24%, respectively, according to the normal value standards of Korean researchers. Furthermore, the prevalence rates of MHE in the three groups of patients with Child-Pugh grades A, B, and C were 52.68%, 58.82%, and 73.53%, respectively, according to the Stroop test standard. However, among the results of each diagnostic standard, the prevalence of MHE showed an increasing trend with an increasing Child-Pugh grade. Further comparison demonstrated that the scores obtained by the number connection test A and the number symbol test were consistent according to the normal value standards of the two studies in China and South Korea ( Z = -0.982, -1.702; P = 0.326, 0.089), while the other three sub-tests had significant differences ( P < 0.001). Conclusion:The prevalence rate of MHE in the cirrhotic population is high, but the prevalence of MHE obtained by using different diagnostic criteria or methods varies greatly. Therefore, in line with the current changes in demographics and disease spectrum, it is necessary to enroll a larger sample size of a healthy population as a control. Moreover, the establishment of more reliable diagnostic scoring criteria will serve as a basis for obtaining accurate MHE incidence and formulating diagnosis and treatment strategies in cirrhotic populations.