ObjectiveTo investigate how Xiaoyao Shukun decoction （XYSKD） regulates the formation and release of neutrophil extracellular traps （NETs） via the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway， thereby reducing inflammation， inhibiting the excessive proliferation of fibroblasts in pelvic adhesion tissue， decreasing adhesion and fibrosis， and repairing the tissue damage in sequelae of pelvic inflammatory disease （SPID）. MethodsA total of 84 Wistar rats were randomly allocated into seven groups： blank， model， XYSKD （8 mg·kg^-1）， mTOR agonist （10 mg·kg^-1）， mTOR agonist + XYSKD （10 mg·kg^-1+8 mg·kg^-1）， mTOR inhibitor （2 mg·kg^-1）， and mTOR inhibitor + XYSKD （2 mg·kg^-1+8 mg·kg^-1）. The rat model of SPID was constructed by starvation， fatigue， and ascending Escherichia coli infection. After 14 days of drug intervention， the ultrastructure of fibroblasts in the pelvic adhesion tissue was observed by transmission electron microscopy. The general morphology of the uterus， fallopian tube， and ovary was observed by laparotomy. The levels of interleukin-1β （IL-1β）， interleukin-17 （IL-17）， and tumor necrosis factor-α （TNF-α） in the peritoneal flushing fluid were determined by enzyme-linked immunosorbent assay （ELISA）. The expression of myeloperoxidase （MPO） and citrullinated histone 3 （H3） in the fallopian tube was detected by immunofluorescence. Western blot and Real-time quantitative polymerase chain reaction （Real-time PCR） were employed to determine the relative protein and mRNA levels， respectively， of neutrophil elastase （NE）， intercellular adhesion molecule-1 （CD54）， α-smooth muscle actin （α-SMA）， H3， PI3K， and Akt. ResultsCompared with the blank group， the model group presented a large number of collagen fibers in bundles， numerous cytoplasmic folds of fibroblasts， reduced or absent mitochondrial cristae， and disordered and expanded endoplasmic reticulum. By laparotomy， extensive pelvic congestion， connective tissue hyperplasia， thickening and hardening of the tubal end near the uterus， and tubal and ovarian adhesion or cyst were observed in the model group. In addition， the model group showed raised levels of IL-1β， IL-17， and TNF-α in the peritoneal flushing fluid （P<0.01）， increased average fluorescence intensities of MPO and H3 （P<0.01）， and up-regulated protein and mRNA levels of NE， H3， CD54， PI3K， and Akt （P<0.01）. Compared with the model group， the mTOR agonist group showed increased fibroblasts and cytoplasmic folds， absence of mitochondrial cristae， endoplasmic reticulum dilation， and evident collagen fiber hyperplasia. Pelvic adhesions were observed to cause aggravated damage to the uterine， fallopian tube， and ovarian tissues. The levels of IL-1β， IL-17， and TNF-α in the peritoneal lavage fluid elevated （P<0.01） and the average fluorescence intensities of MPO and H3 enhanced （P<0.01） in the mTOR agonist group. In contrast， the XYSKD group and the mTOR inhibitor group showcased decreased fibroblasts and collagen fibers， alleviated mitochondrial crista loss and endoplasmic reticulum dilation， improved morphology and appearance of the uterine， fallopian tube， and ovarian tissues， lowered levels of IL-1β， IL-17， and TNF-α in the peritoneal lavage fluid （P<0.01）， decreased average fluorescence intensities of MPO and H3 （P<0.01）， and down-regulated protein and mRNA levels of NE， H3， CD54， PI3K， and Akt （P<0.05）. Compared with the mTOR agonist group， the mTOR agonist + XYSKD group showed alleviated pathological changes in the pelvic tissue， declined levels of IL-1β， IL-17， and TNF-α （P<0.01）， decreased average fluorescence intensities of MPO and H3 （P<0.01）， and down-regulated protein levels of NE， H3， CD54， α-SMA， p-PI3K/PI3K， and p-Akt/Akt （P<0.01） and mRNA levels of NE， H3， CD54， α-SMA， PI3K， and Akt （P<0.01）. Compared with the mTOR inhibitor group， the mTOR inhibitor + XYSKD group demonstrated reduced pathological severity of the pelvic tissue， reduced levels of IL-1β， IL-17， and TNF-α （P<0.01）， decreased average fluorescence intensities of MPO and H3 （P<0.01）， and down-regulated protein and mRNA levels of NE and CD54 （P<0.05）. ConclusionXYSKD can inhibit the excessive formation and release of NETs via PI3K/Akt/mTOR to ameliorate the inflammatory environment and reduce fibrosis and adhesion of the pelvic tissue， thereby playing a role in the treatment of SPID. It may exert the effects by lowering the levels of IL-1β， IL-17， and TNF-α and down-regulating the expression of NE， H3， CD54， α-SMA， PI3K， and Akt in the pelvic adhesion tissue.

Abstract: Objective To conduct a molecular epidemiological tracing and analysis of Yersinia pestis strains isolated from two human plague outbreaks with unknown sources in Gansu Province, China. The results of this analysis would provide a basis for isolating and controlling the sources of Yersinia pestis. Methods The strains of Yersinia pestis isolated from two human plague outbreaks occurring on December 12, 2017, and September 27, 2019 were genotyped by the different region (DFR) and the clustered regularly interspaced short palindromic repeats (CRISPR). The repeat numbers of the variable number tandem repeat (VNTR) loci in the tested strains of Yersinia pestis were calculated by the multiple variable number tandem repeats analysis (MLVA), and the location of the phylogenetic tree of the tested strains was determined with the method of minimum spanning tree (MST) by the software BioNumerics 6.6. Results The strain of 20171212 lacked DFR01, DFR02, DFR03, DFR04, DFR13, DFR23, and the DFR type was identified as type 8. The space sequence of YPa was a1'-a2-a3-a4-a5-a6-a7-a35, the space sequence of YPb was b1-b2-b3-b4, the space sequence of YPc was c1-c2-c3, the gene cluster of CRISPR was Ca35', the genotype of CRISPR was 26'. MLVA clustering analysis showed that the strain clustered within in the cluster of Yuerhong pasture in Subei County and formed an independent branch. On the other hand, the strain of 20190927 lacked DFR01, DFR13 and DFR23, with the DFR type identified as type 1b. The space sequence of YPa was a1-a2-a3-a4-a5-a6-a7, the space sequence of YPb was b1-b2-b3-b4, the space sequence of YPc was c1-c2-c3, the gene cluster of CRSIPR was Ca7, the genotype of CRSIPR was 22 MLVA clustering analysis showed that the strain was located close to the cluster of Dangjinshan in Akesai County, and relatively distant from the cluster of Yuerhong pasture in Subei County. Conclusions The genotypes of strain 20171212 by DFR and CRISPR were consistent with the main genotypes of Y. pestis from Himalayana Marmota foci in Subei County, which confirmed that the human plague cases were naturally occurring locally. However, the strain gathered the cluster of Yuerhong pasture in Subei County, which indicated that the source of infection was not in Yanchiwan Town, but in the surrounding area of the Yuerhong pasture. The genotypes of strain 20190927 by DFR and CRISPR were in accordance with the main genotype of Y. pestis from Himalayana Marmota foci in Akesai County and were closer to the cluster of Dangjinshan in Aksai County than to

Objective:To evaluate the predictive value of the serum N-terminal pro-B-type natriuretic peptide (NT-proBNP) concentration for postoperative acute kidney injury (AKI) in neonates undergoing cardiac surgery.Methods:Perioperative data of 110 consecutive neonates (≤28 days) who underwent cardiac surgery in our hospital from October 2017 to May 2021, were collected retrospectively.According to pROCK criteria, the patients were divided into AKI group and non-AKI group.Demographics, predominant diagnosis, laboratory examination, perioperative management and postoperative outcomes were compared between two groups.The concentration of serum NT-proBNP was routinely measured within 12 h after operation.Multivariate logistic regression analysis was performed for the association between serum NT-proBNP and postoperative AKI.Receiver operating characteristic curve was drawn, and the predictive value of serum NT-proBNP for postoperative AKI was determined according to the area under the curve.Results:A total of 106 neonates were enrolled, and the incidence of postoperative AKI was 54.7%.There were significant difference in the baseline hemoglobin concentration, hematocrit and serum creatinine and serum NT-proBNP concentration between AKI group and non-AKI group ( P<0.05). Multivariate logistic regression analysis indicated that NT-proBNP level was an independent risk factor for AKI after cardiac surgery in neonates ( odds ratio 2.49, 95% confidence interval 1.183-5.23, P=0.016). The area under the curve of NT-proBNP predicting AKI after cardiac surgery was 0.66 (95% confidence interval 0.56-0.76, P=0.007). Conclusions:Elevated serum NT-proBNP concentration is an independent risk factor for AKI after cardiac surgery in neonates and has a certain predictive value for AKI, and close monitoring of perioperative NT-proBNP level is helpful for early identification of high-risk neonates.

Objective:To investigate the infection status of Yersinia in the main host animals of plague in Xiahe and Luqu counties, the Himalayan marmot plague foci of Gansu Province, and to provide a basis for exploring the epidemic status of plague in these foci. Methods:Samples of the ileocecal region and contents, pharyngeal swabs (or tongue roots), and blood of the main host animals of plague in Xiahe County and Luqu County where the plague were active in the 1950s and 1960s were collected from 2014 to 2018. The Yersinia isolation, virulence determination and F1 antibody detection were performed, respectively. Results:Totally 24 strains of Yersinia were detected in 958 samples of ileocecal region and contents with a bacterial detection rate of 2.51%, which were 13 strains of Yersinia enterocolitia (Y.e), 1 strain of Yersinia kristensenii (Y.k), 2 strains of Yersinia frederiksenii/ intermedia (Y.f/i), 6 strains of Yersinia intermedia (Y.i), 1 strain of Yersinia aldouae (Y.a) and 1 strain of Yersinia massiliensis (Y.m). Totally 19 strains of Yersinia were detected in 958 samples of pharyngeal swabs (or tongue roots), and the detection rate was 1.98%, which were 8 strains of Y.e, 1 strain of Yersinia pseudotuberculosis (Y.p), 4 strains of Y.k, 1 strain of Y.f/i, 4 strains of Y.i, and 1 strain of Yersinia ruckeri (Y.r). The virulence types of 21 strains of Y.e were ail -ystA -ystB +yadA -virF -rfbc -, ail -ystA -ystB -yadA -virF -rfbc -, respectively, accounting for 9.52% (2/21) and 90.48% (19/21), none were pathogenic. The results of F1 antibody in 1 079 serum samples were all negative. Conclusions:Yersinia are widely found in the pharynx and intestines of the main host animals of plague in Xiahe and Luqu counties, and the Y.e detected are all non-pathogenic strains. The results of this investigation can provide clues for further study on the preservation of Yersinia pestis in host animals and their living environment.

Objective:To summarize the clinical characteristics, diagnosis and treatment of caspase recruitment domain-containing protein 9 (CARD9) gene deficiency associated invasive candidiasis, and report a novel mutation in CARD9 gene.Methods:The clinical characteristics, laboratory tests, treatment and the outcome of follow-up in a boy with invasive candidiasis were described. The boy′s main clinical manifestations were central nervous system infection and retroperitoneal mass. Whole-exome sequencing was performed and Sanger sequencing was verified to identify the CARD9 gene mutations in the patient and his parents. A literature search for “CARD9”and “invasive candidiasis”was conducted in PubMed, Wanfang and CNKI databases from their establishment to May 2020.Results:A 10-year-old boy suffered onset symptom of chronic diarrhea, which lasted for two months. The symptom was followed by progressive neurological symptoms such as headache, vomiting, seizures and disorder of consciousness. His unusual medical history was absent. Candida albicans were cultured several times in cerebrospinal fluid and blood, and yeast-like fungi were found in the stool high power field of vision. Cerebral magnetic resonance imaging indicated obstructive hydrocephalus and abdominal CT scan showed retroperitoneal mass and thickening of the intestinal wall. The whole-exome sequencing analyses of blood samples from the boy and his parents were performed. The results showed that there was a homozygous mutation of c.952-12_956delinsAG in the CARD9 gene, which was an unreported pathogenic mutation. This was confirmed by Sanger sequencing. There was no significant relief from intravenous combined antifungal medications. After lateral ventricular drainage surgery and injection of amphotericin B into the lateral ventricle, improvement of clinical symptoms and cerebral spinal fluid abnormalities was observed after nine weeks, and the retroperitoneal mass shrank. At follow-up after four-month oral combined antifungal medications, the child had no complaint except fatigue. However, cerebral spinal fluid analysis showed increased protein level and decreased glucose. Persistent hydrocephalus and periventricular white matter abnormal signals were revealed on the brain magnetic resonance imaging and the smaller retroperitoneal mass than before on the abdominal CT scan. In addition to this case, totally 21 cases with CARD9 gene deficiency associated invasive candidiasis have been reported worldwide, most of which featured central nervous system infections.Conclusions:CARD9 gene deficiency is an autosomal recessive primary immunodeficiency that confers human susceptibility to fungal disease. The associated invasive candidiasis often affects the central nervous system and makes the patient severely ill. Adequate systemic antifungal therapies should be given, and patients with hydrocephalus need surgical treatment. A novel mutation is reported that expands the variant diversity of CARD9 gene. For patients with unexplained invasive candidiasis, including those without a history of previous recurrent infection, genetic testing is recommended for primary immunodeficiency including CARD9 gene deficiency.

Objective: To explore the application value of B-mode ultrasound combined with real-time color doppler ultrasound in percutaneous nephrolithotomy and provide guidance for clinical application. Methods: A total of 150 patients underwent percutaneous nephrolithotomy from December 2015 to December 2017 in the People's Hospital of Lishui were selected.According to different ultrasound guidance methods, the patients were divided into two groups.The single group(70 cases) received B-guided puncture.In the combined group(80 cases), B-ultrasound combined with real-time color doppler ultrasound-guided puncture was applied.The incidence of complications and the success rate of lithotomy were compared between the two groups.The changes in renal artery blood flow parameters[end diastolic velocity(EDV), peak systolic velocity(PSV) and resistance index(RI)] before and after surgery in the combined group were observed. Results: The incidence of complications in the combined group was 2.50%(2/80), which was lower than that in the single group[14.29%(10/70)](χ²=7.046, P<0.05). The success rate of stone extraction in the combined group was 98.75%(79/80), which was higher than that in the single group[85.71%(60/70)](χ²=9.336, P<0.01). The EDV and PSV of the renal interlobar arteries of the combined group before and after surgery had statistically significant differences (t=3.794, 5.385, all P<0.05), but the RI had no statistically significant difference (P>0.05). The EDV and PSV of renal segment arteries in the combined group before and after surgery had statistically significant differences (t=4.535, 4.884, all P<0.05), while the RI had no statistically significant difference (P>0.05). The EDV and PSV of renal aorta of the combined group before and after surgery showed no statistically significant differences (all P>0.05), while the RI showed statistically significant difference (t=4.360, P<0.05). Conclusion B-mode ultrasound combined with real-time color doppler ultrasound guidance for percutaneous nephrolithotomy can help reduce the incidence of complications and improve the success rate of stone extraction to a certain extent.

Objective To investigate the relationship between fleas density of Spermophilus alaschanicue and host density as well as meteorological factors.Methods The densities of fleas of Spermophilus alaschanicue and meteorological temperature,precipitation,relative humidity data in the same period from 1977 to 2014 in plague natural foci of Spermophilus alaschanicue of Gansu Province were collected.In order to further establish the multiple linear regression and the stepwise regression analysis model,SAS 9.3 software was used to explore the linear correlation analysis between fleas index number,density of Spermophilus alaschanicue and meteorological factors.Results The average body flea index positively correlated with the density of Spermophilus alaschanicue (r =0.34,P ＜ 0.05),in addition,hole fleas index was in negative correlation with the average temperature of the current and preceding first,second and third years (r =-0.44,-0.39,-0.45,-0.54,P ＜ 0.01).However,hole fleas index positively correlated with the average rainfall of preceding first year (r =0.38,P ＜ 0.05).More specifically,the monthly body flea index was negatively correlated with the average rainfall and the average temperature of current,preceding first,second,and third months,and the average relative humidity of current month (r =-0.23,-0.34,-0.43,-0.29,-0.25,-0.33,-0.36,-0.38,-0.26,P ＜ 0.01).After multiple linear regression analysis,the following equations were obtained:body fleas index =3.983 1 + 0.695 1 × Sperrnophilus density-0.004 9 × average annual rainfall;hole fleas index =10.780 4 + 0.009 1 × average annual rainfall-0.216 2 × annual average relative humidity.Conclusions The relationship between fleas density of Spermophilus alaschanicue and the environment is complex,they have mutual connection as well as mutual restriction.Except Spermophilus density affects the number of fleas,the two most important meteorological factors which affect the flea body index are temperature and rainfall.In addition,the relationship between meteorological factors and flea body index shows somewhat lagging behind for two to three months.

Objective The visual inspection method were not appropriate to perform a hemolysis evaluation for colored injection like doxorubicin hydrochloride,this article adopted three methods to evaluate the hemolysis test of doxorubicin hydrochloride in vitro and provide reference for clinical drug safety.Methods Using rabbit erythrocytes as experimental object,the durg concentration 4.0 and 2.0 mg/mL was chosen which range of clinical concentration and preclinical safety evaluation concentration,to evaluate the hemolysis test of doxorubicin hydrochloride injection with blood analyzer test,direct colorimetric assay,and indirect colorimetric assay.Results The evaluation results of three different methods were very consistent.The tube's hemolysis rate of 4.0 mg/mL dose was far greater than 5％,which means serious hemolysis;Only 0.1 mL tube of 2.0 mg/mL dose (according to the drug concentration equal to 0.5 mL tube of 0.4 mg/mL drug concentration) without hemolysis occurring,the other tubes' hemolysis rates were far greater than 5％,which means serious hemolysis.Conclusion The hemolysis phenomenon may occur when 2.0 mg/mL dose of doxorubicin hydrochloride solution for iv injection is used in clinic and dilution (final concentration not more than 0.4 mg/mL) is recommended.

Objective To type Yersinia pestis isolated from Gansu Province,and to study the trend of diffreent strains in different administrative regions and different years.Methods Totally 193 strains were enrolled in this study,including 9 strains of Ganning Dauricus type,18 strains of Aerjin type,45 strains of Qilian type and 121 strains of Qingzang type.These strains were genotyped by clustered regularly interspaced short palindromic repeats (CRISPR),and genotypes were named according to international standard.Genotyping by CRISPR in different administrative regions and different years of Gansu Province was explored.Results Two clusters (Ca7 and Cb4),including four genotypes (genotypes 7,22,24 and 26) were classified by CRISPR.From the point of view of origin,genotype 24 was the main genotype in Akesai 36.36％ (16/44),Subei 36.17％ (17/47),Yumen 50.00％ (5/10) and Su'nan 38.67％ (29/75);the main genotype of Xiahe and Huining was genotype 26 (4/7);the main genotype of Shandan was genotype 22 (1/1).From the point of view of time,the main genotype of Yersiniapestis in Gansu Province during the years of 1960-1969,1970-1979 and 1980-1989 was genotype 26 [53.33％ (8/15),60.00％ (6/10) and 48.28％ (14/29)];the main genotype was genotype 22 [40.91％ (18/44)] during the years of 1990-1999;and the main genotype was genotype 24 [43.16％ (41/95)] during the years of 2000-2009.Conclusion Four genotypes of Yersiniapestis in Gansu Province are quite different in different administrative regions and different years.

Objective:To observe the therapeutic effects of Tongyushu capsules on dysmenorrhea. Methods:With the intravenous injection of oxytocin in rats, the influence of Tongyushu capsules on rat uterine smooth muscle was observed in vivo. With the intraper-itoneal injection of oxytocin to replicate the mouse model of dysmenorrhea, the effect of Tongyushu capsules on dysmenorrhea was ob-served. Using acetic acid writhing method, the effect of relieving pain Tongyushu capsules in mice was observed. By the method of ear swelling induced by xylene in mice, the anti-inflammatory effect of Tongyushu capsules was observed. By the method of ice water bath and subcutaneous injection of adrenaline, the influence of Tongyushu capsules on the acute blood stasis model in rats was observed. Results:Compared with those of the model group, the contraction of uterine smooth muscle of the groups treated with Tongyushu cap-sules at various dosage was decreased obviously (P<0. 05), the number of body torsion reaction induced by oxytocin and acetic acid in mice was obviously decreased in 15 min (P<0. 05), the degree of ear swelling in mice treated with Tongyushu capsules was signifi-cantly reduced (P<0. 05), and the whole blood viscosity, plasma viscosity, hematokrit, erythrocyte sedimentation rate and erythro-cyte electrophoresis time were significantly decreased (P<0. 05). Conclusion:Tongyushu capsules have obvious effects of the inhibi-tion of excessive contraction of uterine smooth muscle, dysmenorrhea alleviation, anti-inflammation and relieving pain, which show good therapeutic effect on dysmenorrhea.