Objective:To analyze the diagnostic value of cell-free plasma metagenomic next-generation sequencing (mNGS) pathogen identification for severe aplastic anemia (SAA) bloodstream infection.Methods:From February 2021 to February 2022, mNGS and conventional detection methods (blood culture, etc.) were used to detect 33 samples from 29 consecutive AA patients admitted to the Anemia Diagnosis and Treatment Center of the Hematology Hospital of the Chinese Academy of Medical Sciences to assess the diagnostic consistency of mNGS and conventional detection, as well as the impact on clinical treatment benefits and clinical accuracy.Results:①Among the 33 samples evaluated by mNGS and conventional detection methods, 25 cases (75.76%) carried potential pathogenic microorganisms. A total of 72 pathogenic microorganisms were identified from all cases, of which 65 (90.28%) were detected only by mNGS. ②All 33 cases were evaluated for diagnostic consistency, of which 2 cases (6.06%) were Composite, 18 cases (54.55%) were mNGS only, 2 cases (6.06%) were Conventional method only, 1 case (3.03%) was both common compliances (mNGS/Conventional testing) , and 10 cases (30.3%) were completely non-conforming (None) . ③All 33 cases were evaluated for clinical treatment benefit. Among them, 8 cases (24.24%) received Initiation of targeted treatment, 1 case (3.03%) received Treatment de-escalation, 13 cases (39.39%) received Confirmation, and the remaining 11 cases (33.33%) received No clinical benefit. ④ The sensitivity of 80.77%, specificity of 70.00%, positive predictive value of 63.64%, negative predictive value of 84.85%, positive likelihood ratio of 2.692, and negative likelihood ratio of 0.275 distinguished mNGS from conventional detection methods (21/12 vs 5/28, P<0.001) . Conclusion:mNGS can not only contribute to accurately diagnosing bloodstream infection in patients with aplastic anemia, but can also help to guide accurate anti-infection treatment, and the clinical accuracy is high.

Objective:To explore the method of ultrasonography for detecting the fetal umbilical vein diameter, blood flow volume and normalized volume blood flow and establish normal reference ranges with umbilical vein diameter, blood flow volume and normalized blood flow and Z-scores for umbilical vein diameter and blood volume flow.Methods:This was a prospective study on 907 normal fetuses in the Second Xiangya Hospital, Central South University and Women and Children Healthcare Hospital of Zhuzhou from March 2019 to December 2020. The umbilical vein diameter (Duv), umbilical vein blood flow volume (Quv) and normalized volume blood flow (nQ = Quv/estimated fetal weight) of the free loop of umbilical vein (FUV) and fetal intra-abdominal umbilical vein (IUV) were collected. And the mean values and 90% confidence intervals of Duv, Quv and nQ in two segments of umbilical veins at different gestational ages were calculated. Regression analysis of Duv, Quv and nQ were performed with gestational age (GA), and the parameters of umbilical vein in different segments were compared. Finally, with gestational age (GA) as the independent variable, Z-scores of the Duv and Quv were built.Results:The mean values and 90% confidence intervals of Duv, Quv, and nQ in 858 (94.6%) normal fetal umbilical veins were successfully obtained. The Duv, Quv of the FUV and IUV increased as pregnancy progressed. The Quadratic curve of Duv and Linear curve of Quv were of the highest fitnesses, respectively( r=0.951, 0.941, 0.986, 0.982; all P<0.001). While nQ increased with GA followed by a decreased trend, and the Quadratic curve was the highest fitting curve of nQ( r=0.610, 0.611; all P<0.001). Duv-FUV was greater than Duv-IUV( P<0.001), nQ-FUV was bigger than Quv-IUV( P=0.001), and he difference was not statistically significant between Quv-FUV and Quv-IUV( P=0.133). Z-scores models of Duv and Quv were successfully established, and all Z-scores were Gaussian distribution. Conclusions:The normal ranges and Z-scores of umbilical vein parameters are useful to improve the evaluation of placental circulation and provide a strong basis for the monitoring of fetus-related diseases and the evaluation of pregnancy prognosis. The choice of FUV or IUV umbilical vein to evaluate placental circulation may depend on the actual situation in clinical application.

ObjectiveTo investigate the effect of Shengjiang Tonglong prescription hollow suppository on rats with prostate hyperplasia， and the effect of the proteins related to phosphoinositide 3-kinase/protein kinase B （PI3K/Akt） signaling pathway in the prostate， thus exploring the mechanism of Shengjiang Tonglong prescription hollow suppository in the treatment of rats with prostate hyperplasia. MethodTen SD male rats were randomly selected from 60 SD male rats to form a sham operation control group， and the rest rats were subcutaneously injected with testosterone propionate for 4 consecutive weeks after castration to induce the rat model of prostatic hyperplasia. According to the random number table method， the 50 rats were randomly divided into a model group， a finasteride group （0.45 mg·kg^-1）， and three high， middle， and low-dose Shengjiang Tonglong prescription hollow suppository groups （3.98， 1.99， 0.99 g·kg^-1）， with ten rats in each group. After castration for 7 d， the sham operation control group and the model group used the blank hollow suppositories， and the finasteride group and the Shengjiang Tonglong prescription hollow suppository groups used the corresponding hollow suppositories. The drugs were given to the rats by anal plugs continuously for 28 d. The rats were then killed， and the prostate tissues were separated and weighed to observe the effects of drugs on the prostate index of rats in each group. The hematoxylin-eosin （HE） staining was used for the pathological observation of the prostate tissues. The level of dihydrotestosterone （DHT） was detected by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the expression levels of PI3K/Akt signaling pathway protein， B-cell lymphoma-2 （Bcl-2）， cysteine aspartate-specific protease-3 （Caspase-3）， Bcl-2-associated X protein （Bax）， and αB-crystallin （CRYAB） protein in the prostate tissues. ResultAs compared with the sham operation control group， the protein expression levels of prostate index， DHT level， CRYAB， Bcl-2， PI3K， and Akt in the model group were increased， and the protein expression levels of Caspase-3 and Bax were decreased （P<0.05， P<0.01）. As compared with the model group， the prostate index in the high-dose Shengjiang Tonglong prescription hollow suppository group was decreased （P<0.05）， and the level of DHT and the protein expression levels of CRYAB， Bcl-2， PI3K， and Akt in the prostate of the Shengjiang Tonglong prescription hollow suppository groups were decreased， and the protein expression levels of Caspase-3 and Bax were increased （P<0.05， P<0.01）. ConclusionShengjiang Tonglong prescription hollow suppository decreases the expression of CRYAB protein， negatively regulates the PI3K/Akt signaling pathway， down-regulates the level of DHT and the protein expression levels of Bcl-2， PI3K， and Akt， and up-regulates the protein expression levels of Caspase-3 and Bax， thereby inhibiting cell proliferation and promoting cell apoptosis， which plays a therapeutic role in the benign prostate hyperplasia （BPH）. The high-dose Shengjiang Tonglong prescription hollow suppository significantly improves prostatic hyperplasia in rats.

ObjectiveTo investigate the effect of Shengjiang Tonglong prescription hollow suppository on rats with prostate hyperplasia， and the effect of the proteins related to phosphoinositide 3-kinase/protein kinase B （PI3K/Akt） signaling pathway in the prostate， thus exploring the mechanism of Shengjiang Tonglong prescription hollow suppository in the treatment of rats with prostate hyperplasia. MethodTen SD male rats were randomly selected from 60 SD male rats to form a sham operation control group， and the rest rats were subcutaneously injected with testosterone propionate for 4 consecutive weeks after castration to induce the rat model of prostatic hyperplasia. According to the random number table method， the 50 rats were randomly divided into a model group， a finasteride group （0.45 mg·kg^-1）， and three high， middle， and low-dose Shengjiang Tonglong prescription hollow suppository groups （3.98， 1.99， 0.99 g·kg^-1）， with ten rats in each group. After castration for 7 d， the sham operation control group and the model group used the blank hollow suppositories， and the finasteride group and the Shengjiang Tonglong prescription hollow suppository groups used the corresponding hollow suppositories. The drugs were given to the rats by anal plugs continuously for 28 d. The rats were then killed， and the prostate tissues were separated and weighed to observe the effects of drugs on the prostate index of rats in each group. The hematoxylin-eosin （HE） staining was used for the pathological observation of the prostate tissues. The level of dihydrotestosterone （DHT） was detected by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the expression levels of PI3K/Akt signaling pathway protein， B-cell lymphoma-2 （Bcl-2）， cysteine aspartate-specific protease-3 （Caspase-3）， Bcl-2-associated X protein （Bax）， and αB-crystallin （CRYAB） protein in the prostate tissues. ResultAs compared with the sham operation control group， the protein expression levels of prostate index， DHT level， CRYAB， Bcl-2， PI3K， and Akt in the model group were increased， and the protein expression levels of Caspase-3 and Bax were decreased （P<0.05， P<0.01）. As compared with the model group， the prostate index in the high-dose Shengjiang Tonglong prescription hollow suppository group was decreased （P<0.05）， and the level of DHT and the protein expression levels of CRYAB， Bcl-2， PI3K， and Akt in the prostate of the Shengjiang Tonglong prescription hollow suppository groups were decreased， and the protein expression levels of Caspase-3 and Bax were increased （P<0.05， P<0.01）. ConclusionShengjiang Tonglong prescription hollow suppository decreases the expression of CRYAB protein， negatively regulates the PI3K/Akt signaling pathway， down-regulates the level of DHT and the protein expression levels of Bcl-2， PI3K， and Akt， and up-regulates the protein expression levels of Caspase-3 and Bax， thereby inhibiting cell proliferation and promoting cell apoptosis， which plays a therapeutic role in the benign prostate hyperplasia （BPH）. The high-dose Shengjiang Tonglong prescription hollow suppository significantly improves prostatic hyperplasia in rats.

Objective:To analyze the causes of unplanned reoperation in pediatric patients after elective digestive tract surgery and the prognosis.Methods:Medical records were reviewed from pediatric patients undergoing unplanned reoperation after elective digestive tract surgery at our department from Jan 2012 to Dec 2019. Primary diagnoses, procedures and levels of index surgeries, causes and procedures of unplanned reoperations, and patients' prognosis were analyzed.Results:There were 39 cases, and the primary diagnoses included biliary disease, anal and colorectal disease, and intestinal disease. There were 4 (10%) cases of level Ⅱ surgeries,and 35 (90%) cases of level Ⅲ&Ⅳ surgeries. The index surgical procedures included 19 (49%) biliary-intestinal procedures, 11 (28%) simple intestinal procedures, and 9 (23%) anal and colorectal procedures. The direct causes of unplanned reoperation included 10 (26%) anastomotic leakages, 8 (20%) adhesive intestinal obstructions, 5 (13%) postoperative intussusceptions, 5 (13%) incisional complications (infection, dehiscence and incisional hernia), 3 (8%) postoperative hemorrhages and 8 (20%) miscellaneous (iatrogenic injury and surgical misjudgment). Patients' prognosis included 24 (62%) full recoveries, 9 (23%) further operations, and 5 (13%) deaths, and 1 (3%) short bowel syndrome.Conclusions:Most pediatric unplanned reoperations after elective digestive tract surgery occur in complex surgical procedures. The most common causes of unplanned reoperation are anastomotic leakage, adhesive intestinal obstruction. Unplanned reoperations are often prone to adverse effects on prognosis.

Objective:To evaluate clinical characteristics and treatment of postoperative anastomotic stricture in pediatric congenital biliary dilatation patients.Methods:The clinical data of 24 children with postoperative anastomotic stricture from Apr 2012 to Oct 2019 in Beijing Children's Hospital was retrospectively analyzed.Results:There were 6 males and 18 females. Patients were divided into bile- leak group (BL, n=6) and non bile-leak group (NBL, n=18) based on whether there was anastomotic leakage after primary surgery. The main symptoms in BL group was persistent obstructive jaundice, and recurrent cholangitis in NBL group. Postoperative symptoms were first shown in an average of 7.0 months in BL group, compared to 59.0 months in NBL group, P<0.05. In BL group, 4 underwent redoing hepaticojejunostomy, 2 underwent anastomosis plasty. In NBL group, 3 underwent redoing hepaticojejunostomy, 15 did anastomosis plasty with multiple biliary stones found necessitating extraction. After reoperation, one patient had bile leakage, 2 patients had recurrent cholangitis within one-month, 21 patients had uneventful recovery. Five were found to have biliary stones in long-term follow-up. Conclusions:Biliary-enteric anastomotic leakage can cause stricture in postoperative patients of congenital biliary dilatation ,reoperation is necessary in symptomatic patients.

Objective To study the trends of lung cancer mortality among adult residents in Macheng City, Hubei Province from 1984 to 2018． Methods Mortality data was extracted from Macheng City disease surveillance points (DSPs) system and China Demographic Yearbook. The age-period-cohort (APC) model and Intrinsic Estimator algorithm were used to estimate the age effect, period effect and cohort effect of lung cancer mortality. Results The age effect coefficient of lung cancer mortality increased with age from 20 to 74 years old. The mortality risk of the 70-74 group was 42.62 times that of the 20-24 group. The period effect coefficient of lung cancer mortality also continued to rise with time. The cohort effect coefficient was parabolic, and residents born in 1939-1943 had the highest coefficient (1.298 4). Conclusion The risk of lung cancer death of adult residents in Macheng City significantly increased with the year and the rapid development of socio-economics.

Objective: To improve the diagnostic criteria of suspected cases through investigating the epidemiological and clinical manifestations of confirmed cases of new-type coronavirus infection in children. Methods: We retrospective analyzed the epidemiological and clinical manifestations of 6 children with new coronavirus infection diagnosed in Chongqing Three Gorges Central Hospital from February 3, 2020 to February 15, 2020 . Compared with the diagnostic criteria of suspected cases,we summarized the problems encountered in the application of this standard in clinical work and try to put forward Suggestions for improvement. Results: Among the 6 children with confirmed cases: 5 males and 1 female; 3 from Hubei Province and 3 from Wanzhou ; 6 cases of clustered onset of the family; Visiting nature: 3 cases of suspected case income, 3 cases of community or outpatient screening . Three cases with fever and / or respiratory symptoms, one of which had symptoms of diarrhea; all children's blood routine and lymphocyte counts were within the normal range; chest CT imaging except for cases No. 1 and No. 5 were in line with typical new coronavirus pneumonia signs. In addition, the remaining 3 patients had abnormal imaging but did not have the characteristics of new coronavirus pneumonia, and 1 case was normal. Comparison results:Only case 1 of all cases fully met the diagnostic criteria, and the remaining cases did not meet the diagnostic criteria of early suspected cases. Conclusion In order to improve the accuracy and practicality of the diagnosis of suspected cases in children, it is recommended to refine and standardize the diagnostic criteria of some suspected cases.

Objective: To investigate the role of the bone morphogenetic protein 2 (BMP2)⁃Smad1/5 and p38MAPK signaling pathways in the osteogenic differentiation of MSMSCs by insulin⁃like growth factor 1 (IGF1). Methods : A re⁃ combinant adenovirus (RAD) and IGF1 expressing IGF1 gene were constructed. After osteogenic induction, qRT⁃PCR and Western blot were used to detect the phosphorylation level of Smad1/5 and the expression of the BMP⁃2 protein in the BMP⁃Smad signaling pathway; immunohistochemistry was used to observe the nuclear translocation of Smad1/5; qRT⁃PCR and Western blot were used to detect IGF with Noggin and SB203580, inhibitors of the p38MAPK signaling path⁃ way 1⁃mediated osteogenic differentiation of MSMSCs Results: The recombinant IGF1 adenovirus was constructed suc⁃ cessfully. MSMSCs were cultured in inductive medium after infection with different concentrations of Ad⁃IGF1, and then, the protein levels of BMP2 and p⁃Smad1/5 increased. IGF1 can also induce nuclear translocation of Smad1/5. In addition, Noggin significantly reduced the phosphorylation level of Smad1/5 and the formation of mineralized nodules in the MSMSCs. The mRNA levels of Runx2, OPN and ALP also decreased. In contrast, SB203580 decreased neither the phosphorylation level of p38 nor the mRNA expression of Runx2, OPN and ALP in the Ad⁃IGF1 MSMSCs Conclu⁃sion IGF1 can promote the osteogenic differentiation of MSMSCs via the BMP2⁃Smad1/5 signaling pathway. In con⁃ trast, IGF1 may not promote the osteogenic differentiation of MSMSCs via the p38MAPK signaling pathway.

Objective:To improve the diagnostic criteria of suspected cases through investigating the epidemiological and clinical manifestations of confirmed cases of novel coronavirus infection in children.Methods:We retrospectively analyzed the epidemiological and clinical manifestations of six children with novel coronavirus infection diagnosed in Chongqing Three Gorges Central Hospital from February 3, 2020 to February 15, 2020.Compared with the diagnostic criteria of suspected cases, we summarized the problems encountering in the application of this standard in clinic and try to put forward suggestions for improvement.Results:Among the six confirmed cases: five males and one female; three from Hubei Province and three from Wanzhou; six cases of clustered onset of the family.Visiting nature: six cases of suspected case, six cases from community or outpatient screening.Three fever cases with/without respiratory symptoms, one of which had diarrhea; all children′s blood routine and lymphocyte counts were within the normal range; chest CT imagings of No.1 and No.5 case showed typical novel coronavirus pneumonia signs, and the other three patients had abnormalities without the characteristics of novel coronavirus pneumonia, and one case was normal.Comparison results: Only No.1 case fully met the diagnostic criteria, and the remaining cases did not meet the diagnostic criteria for early suspected cases.Conclusion:In order to improve the accuracy and practicality of the diagnosis of suspected cases in children, it is recommended to refine and standardize the diagnostic criteria for some suspected cases.