Mycobacterium intracellular CHPC 1.5701 strain was isolated from the sputum specimen of the patient.This study attempted to establish a guinea pig infection model to evaluate its virulence,so as to provide basic scientific basis for the prevention and treatment of Mycobacterium intracellular infection.Mycobacterium intracellular CHPC 1.5701 was cultured in 7H10 medium.The morphology of the colony changed from colorless to light yellow smooth colony,and the acid-fast staining was positive.Mycobacterium intracellular suspension was diluted with 0.9％sodium chloride solution to 1 × 1010 CFU/mL,1×109 CFU/mL,1 × 108 CFU/mL,1 × 107 CFU/mL and 1 × 106 CFU/mL,respectively.Healthy Hartly guinea pigs with negative skin test of pure protein derivatives of tuberculin were selected and randomly divided into 6 groups with 10 guinea pigs in each group,half male and half female.Guinea pigs in the ex-perimental group were divided into 5 groups,which were intra-peritoneally injected with 5 different concentrations of 1.0 mL suspension/guinea pig,and those in the control group were in-traperitoneally injected with 0.9％sodium chloride solution 1.0 mL suspension/guinea pig.The guinea pigs was then observed,weighed every week,dissected 5 weeks later,lung,spleen and liver tissues were taken,and the tissue lesions were analyzed by hematoxylin-eosin(HE)staining,and the bacteria load was detected by tissue homogenate culture in 7H10 medium.The results showed that Mycobacterium intracellular challenge test had no significant effect on body weight of male guinea pigs(P＞0.05),but could reduce body weight of female guinea pigs(P＜0.01).No pathological changes of organ tissues were ob-served in guinea pigs in the control group,while HE staining of lung,spleen and liver tissues of guinea pigs in challenge experi-mental groups showed alveolar wall alveolar epithelial hyperplasia,monocyte infiltration in liver and spleen,granulomatous in-flammation and other pathological changes,and the degree of pathological changes was related to the injection dose of Mycobac-terium intracellular.Mycobacteria intracellular could be isolated from tissue culture of some organs,among which the spleen had a large amount of bacteria.Intrabitoneal injection of Mycobacteria intracellular CHPC 1.5701 can affect the body weight of female guinea pigs and cause lung,liver and spleen infection of guinea pigs,but no caseous necrosis of guinea pigs was ob-served.Compared with the reported data of Mycobacterium tuberculosis virulence test,it is concluded that Mycobacterium in-tracellular has certain virulence to guinea pigs and belongs to low virulence and low pathogenicity bacteria...

Mycobacterium intracellular CHPC 1.5701 strain was isolated from the sputum specimen of the patient.This study attempted to establish a guinea pig infection model to evaluate its virulence,so as to provide basic scientific basis for the prevention and treatment of Mycobacterium intracellular infection.Mycobacterium intracellular CHPC 1.5701 was cultured in 7H10 medium.The morphology of the colony changed from colorless to light yellow smooth colony,and the acid-fast staining was positive.Mycobacterium intracellular suspension was diluted with 0.9％sodium chloride solution to 1 × 1010 CFU/mL,1×109 CFU/mL,1 × 108 CFU/mL,1 × 107 CFU/mL and 1 × 106 CFU/mL,respectively.Healthy Hartly guinea pigs with negative skin test of pure protein derivatives of tuberculin were selected and randomly divided into 6 groups with 10 guinea pigs in each group,half male and half female.Guinea pigs in the ex-perimental group were divided into 5 groups,which were intra-peritoneally injected with 5 different concentrations of 1.0 mL suspension/guinea pig,and those in the control group were in-traperitoneally injected with 0.9％sodium chloride solution 1.0 mL suspension/guinea pig.The guinea pigs was then observed,weighed every week,dissected 5 weeks later,lung,spleen and liver tissues were taken,and the tissue lesions were analyzed by hematoxylin-eosin(HE)staining,and the bacteria load was detected by tissue homogenate culture in 7H10 medium.The results showed that Mycobacterium intracellular challenge test had no significant effect on body weight of male guinea pigs(P＞0.05),but could reduce body weight of female guinea pigs(P＜0.01).No pathological changes of organ tissues were ob-served in guinea pigs in the control group,while HE staining of lung,spleen and liver tissues of guinea pigs in challenge experi-mental groups showed alveolar wall alveolar epithelial hyperplasia,monocyte infiltration in liver and spleen,granulomatous in-flammation and other pathological changes,and the degree of pathological changes was related to the injection dose of Mycobac-terium intracellular.Mycobacteria intracellular could be isolated from tissue culture of some organs,among which the spleen had a large amount of bacteria.Intrabitoneal injection of Mycobacteria intracellular CHPC 1.5701 can affect the body weight of female guinea pigs and cause lung,liver and spleen infection of guinea pigs,but no caseous necrosis of guinea pigs was ob-served.Compared with the reported data of Mycobacterium tuberculosis virulence test,it is concluded that Mycobacterium in-tracellular has certain virulence to guinea pigs and belongs to low virulence and low pathogenicity bacteria...

Objective:To explore the material basis and mechanism of Sangjiang Ganmao injection （SG） in the treatment of common cold by ultra-performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry （UPLC-Q-Orbitrap HRMS） and network pharmacology. Method:UPLC-Q-Orbitrap HRMS was used to identify the chemical components of SG with mobile phase of acetonitrile （A）-0.1% formic acid aqueous solution （B） for gradient elution （0-10 min， 4%-15%A； 10-35 min， 15%-30%A； 35-45 min， 30%-33%A； 45-55 min， 33%-60%A； 55-58 min， 60%A）， flow rate of 0.2 mL·min^-1， electrospray ionization （ESI） and scanning range of m/z 100-1 500 under positive and negative ion modes. Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and GeneCards 5.0 database were used to screen and predict the potential targets of chemical components in SG， STRING 11.0 database and Cytoscape 3.7.2 software were used to construct protein-protein interaction （PPI） network model， gene ontology （GO） analysis and pathway analysis were performed on potential targets by Metascape 3.5， Reactome database and Kyoto Encyclopedia of Genes and Genomes （KEGG）， Cytoscape 3.7.2 software was used to build the network of "herbs-ingredients-key targets". Result:A total of 54 components in SG were identified， and 80 potential targets of SG for treatment of common cold were predicted and screened based on this. SG exerted therapeutic effects by acting on targets such as interleukin （IL）-6， tumor necrosis factor （TNF） and IL-10， and signaling pathways such as IL-17 signaling pathway， TNF signaling pathway and interaction of cytokine receptors. Conclusion:SG may interfere with the expression of inflammatory cytokines by acting on related targets and pathways such as inflammation and immune system， and regulate the immune function of the body as a whole， thereby exerting a therapeutic effect.

italic>Dendrobium officinale Kimura et Migo is a rare Chinese herbal medicine, while Dendrobium crepidatum Lindl is a local medicine in Yunnan, both of which have the function of nourishing yin and stomach. To reveal the differences in chemical composition between the two species, ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-Q-TOF-MS/MS) was used to analyze the chemical composition of stems and leaves of D. officinale and D. crepidatum. Principal component analysis (PCA) and partial least squares discriminant analysis (OPLS-DA) were used to determine the differences in metabolites between species and parts of Dendrobium. Fifty-eight chemical compounds were identified in the two species. Analysis indicated that the side ring of alkaloids connected with nitrogen was readily cleaved during analysis. The results of PCA analysis showed that the stems and leaves of D. officinale and D. crepidatum could be easily differentiated, and the chemical constituents of D. officinale and D. crepidatum were significantly different. OPLS-DA analysis showed that there were 16 metabolite differences between the stems and 22 differences in metabolites between the leaves of D. officinale and D. crepidatum. The main metabolite differences in components between the two Dendrobium species were dendrocrepidine B, dendrocrepidine C and dendrocrepine. There were 14 differences in metabolites between the stems and leaves of D. crepidatum. In conclusion, the chemical compositions of D. officinale and D. crepidatum are quite different; the small molecular compounds of D. officinale are mainly terpenoids and flavonoids, and the content of alkaloids is low. There is no significant difference between stem and leaf. In contrast, D. crepidatum is mainly composed of alkaloids and terpenoids, with crepidamine and dendrocrepine as its unique components, and there are great differences in the components between stems and leaves. This study provides a theoretical basis for the development and utilization of Dendrobium resources.

To explore the optimal therapy time for the treatment of severe coronavirus disease 2019(COVID-19)by traditional Chinese medicine(TCM)and its influence on the therapeutic effect and prognosis. The clinical data,laboratory findings,and outcomes of 64 patients with severe COVID-19 treated with TCM and western medicine in Chongqing from January 20,2020, to March 11,2020 were retrospectively analyzed.Patients were divided into early intervention group[TCM was initiated within 3 days (including day 3) after the first diagnosis of severe type/critical type COVID-19]and late intervention group[TCM was initiated after 7 days (including day 7) after the first diagnosis of severe type /critical type COVID-19].The changes in clinical parameters during the course of disease were compared between the two groups. On day 14,the oxygenation index was 292.5(252.0,351.0)mmHg in the early intervention group,which was significantly higher than that in the late intervention group [246.0(170.0,292.5)mmHg](=0.005).The length of hospital stay [(18.56±1.11)d (24.87±1.64)d,=0.001],duration of ICU stay [(14.12±0.91)d (20.00±1.53)d,=0.000] and time to negativity [(16.77±1.04)d (22.48±1.66)d,=0.001] in the early intervention group were significantly shorter than those in the late intervention group.The intubation rate(7.3%)in the early intervention group was significantly lower than that in the late intervention group(30.4%)(=0.028). Early TCM therapy within three days after a diagnosis of severe COVID-19 can shorten the length of hospital stay,duration of ICU stay,and time to negativity and decrease intubation rate.
Betacoronavirus ; Coronavirus Infections ; drug therapy ; Humans ; Medicine, Chinese Traditional ; Pandemics ; Pneumonia, Viral ; drug therapy ; Prognosis ; Retrospective Studies

Objective:To observe the effect of early weight-bearing on the appropriate population with intertrochanteric fracture after surgery. Methods:From April, 2017 to December, 2018, a total of 45 patients with Evans-Jensen type II intertrochanteric fracture and fracture reduction as positive medial cortex support (PMCS) after proximal femoral nail anti-rotation (PFNA) fixation were randomly divided into control group (n = 22) and experimental group (n = 23). Weight-bearing as tolerated (WBAT) was initiated from six weeks after surgery in the control group, and within 48 h after surgery in the experimental group. The frequency of WBAT in two groups increased gradually from three times a day for ten minutes a time to five times a day for 20 minutes a time until clinical healing of fracture. The length of stay, hospital cost, the fracture healing time and the complication incidence were compared between two groups, as well as the scores of Visual Analogue Scale (VAS) and Harris Hip Score at six weeks, three months and six months after surgery. Results:Compared with the control group, the length of stay was shorter (t = 3.716, P < 0.01), the hospital cost was lower, but no significant difference was found (t = 1.540, P > 0.05), and the fracture healing time was shorter (t = 6.248, P < 0.001) in the experimental group. The complication incidence was lower in the experimental group, but there was no significant difference (χ²= 2.198, P > 0.05). Six weeks, three months and six months after surgery, there was no significant difference in the score of VAS between two groups (t < 1.330, P > 0.05). The score of Harris Hip Score was significantly higher in the experimental group than in the control group six weeks after surgery (t = -5.115, P < 0.001), however, no significant difference was found in other time points (|t| < 1.799, P > 0.05). Conclusion:Early weight-bearing within 48 h after PFNA fixation for Evans-Jensen type II intertrochanteric fractures and reduction with PMCS could shorten the length of stay, shorten the bony healing time and promote early recovery of hip function.

OBJECTIVE To improve the poor water solubility and evaluating poor acitivity of etoposide (VP-16) by preparing VP-16 nanoparticle suspension (VP-16 NP) and its penetration through the blood-brain barrier (BBB).METHODS VP-16 NP was prepared with the anti-solution exchange method.The shape structure and diameter were observed with transmission electron microscopy (TEM) and scanning electron microscopy (SEM) and dynamic light scattering (DLS), respectively. The drug release profiles of the VP-16 powder and VP-16 NP were measured.The effect of VP-16 NP on the growth of KB cells was observed via MTT assay. In addition, primary brain microvascular endothelial cells from 1stto 2nd generation of SD rats at two weeks of age were used to construct an in vitro BBB model.The classic 4 h leak test,trans-epithelial electrical resistance (TEER) test and fluorescein disodium salt(FLU)perme?ability test were conducted to verify whether the in vitro BBB model was successfully established.RESULTS VP-16 NP was a solid sphere with a size of 140 nm detected by TEM,SEM and DLS.The cumulative release rate of VP-16 NP was 3 times that of VP-16 powder. The results of MTT colorimetric assay showed that VP-16 powder had no inhibitory effect on KB cells,while VP-16 NP could effectively inhibit KB cells. In the 4 h leakage experiment, the top and bottom chambers of the Transwell cell model could maintain a liquid surface distance of >0.5 cm,indicating that the in vitro BBB model was initially formed.The effective resistance value of the TEER experiment was 223 Ω·cm2,indicating that the in vitro BBB model was basically established. In FLU permeability experiments, the permeability coefficients were respectively (0.33±0.04)×10-3,(0.42±0.07)×10-3,and (0.52±0.06)×10-3cm·min-1at 15,30 and 60 min, indicating that the model had low permeability.The above three experiments suggested that the BBB in vitro model was successfully constructed. On this basis, the in vitro BBB model was used to evaluate the penetration of VP-16 NP at a permeability coefficient of (1.87±0.03)×10-3cm·min-1at 30 min,showing high permeability.VP-16 NP showed better penetration of BBB.CONCLUSION VP-16 NP is success?fully prepared,which increases the release rate of the drug,enhances the killing effect of the cells,and shows good penetration through the in vitro BBB model evaluation.

Objective The prognostic expression level and prognostic significance of CX3CL1 in patients with non-small cell lung cancer(NSCLC) need further investigation. The purpose of this paper was to investigate the effects of various CX3CL1 mRNA expression levels on patients with NSCLC.Methods By retrieving lung-cancer related gene expression profile data in NCBI GEO database and TCGA of UCSC Cancer Browser, 8 datasets were included based on inclusion and exclusion criteria. All the datasets were collated and standardized through R statistical software. Univariate and multivariate Cox models were conducted for prognosis analysis of CX3CL1 in each dataset. HR values of all the datasets were pooled by meta algorithm.Results High-expression of CX3CL1 mRNA in tumor tissues of lung adenocarcinoma was a positive prognostic factor for overall survival(pooled HR=0.53; 95% CI=0.43-0.65 in univariate analysis; pooled HR=0.52; 95% CI=0.42-0.64 in multivariate analysis). However, in lung squamous cell carcinoma, there was no significant association between CX3CL1 expression and overall survival (pooled HR=1.09; 95% CI=0.82-1.45 in univariate analysis; pooled HR=1.18; 95% CI=0.88-1.58 in multivariate analysis).Conclusion The mRNA level of CX3CL1 in lung adenocarcinoma was positively correlated with better prognosis, but there was no correlation between CX3CL1 mRNA level and prognosis in patients with lung squamous cell carcinoma. CX3CL1 may be used as a potential prognostic marker for patients with lung adenocarcinoma.

OBJECTIVETo observe the effect of Trichinella spiralis and its worm-derived proteins on cecal ligation and puncture (CLP)-induced sepsis in mice.

METHODSEighty male BALB/c mice were randomly divided into sham-operated group, CLP group, Trichinella spiralis muscle larvae (ML) pre-infection group (ML+CLP group), soluble muscle larvae proteins (SMP) treatment group (SMP+CLP group) and excretory-secretory proteins (MES) treatment group (MES+CLP group). In ML+CLP group, the mice were orally infected with 300 Trichinella spiralis muscle larvae at 28 days before CLP and those in the other groups were intraperitioneally injected with PBS or SMP (25 µg/mice) or MES (25µg/mice) 30 min after CLP. The general condition and 72-h survival after CLP of the mice were observed. The levels of alanine transaminase (ALT), aspartate transaminase (AST), blood urea nitrogen (BUN), creatinine (Cr), TNF-α, IL-6, IL-1β, IL-10 and TGF-β were measured at 12 h after the operation, and the pathological changes of the liver and kidney were observed.

RESULTSs Compared with the sham-operated mice, the mice in CLP group showed decreased 72-h survival, obviously increased ALT, AST, BUN, Cr, TNF-α, IL-6, IL-1β, IL-10, and TGF-β with hepatic cords disorder, hepatocytes swelling, glomerulus shrinkage, and renal tubular cell edema. Compared with CLP group, the mice in ML+CLP group showed lowered levels of ALT, AST, Cr, TNF-α and IL-1β and increased levels of IL-10 and TGF-β; in SMP+CLP group, the levels of ALT, AST, Cr, TNF-α and IL-1β were decreased and TGF-β increased. In MES+CLP group, the mice showed obviously increased 72-h survival with lowered levels of ALT, AST, BUN, Cr, TNF-α, IL-6 and IL-1β, increased levels of IL-10 and TGF-β, and alleviated liver and kidney damages.

CONCLUSIONTrichinella spiralis and its worm-derived proteins can decrease the levels of pro-inflammatory cytokines and increase immunomodulatory cytokines, and MES has more potent effect to reduce structural and functional damages of the liver and kidney.

Alanine Transaminase ; blood ; Animals ; Antigens, Helminth ; pharmacology ; Aspartate Aminotransferases ; blood ; Blood Urea Nitrogen ; Cecum ; Creatinine ; blood ; Cytokines ; blood ; Helminth Proteins ; pharmacology ; Kidney ; physiopathology ; Kidney Diseases ; therapy ; Ligation ; Liver ; physiopathology ; Liver Diseases ; therapy ; Male ; Mice ; Mice, Inbred BALB C ; Sepsis ; therapy ; Trichinella spiralis

OBJECTIVETo investigate the clinical application of free superficial iliac circumflex artery skin flaps, as well as the management of donor site defects.

METHODS17 free superficial iliac circumflex artery skin flaps were applied for the traumatic defects or deformities on face, neck, foot, hand, ankle and lower leg, respectively. The donor site defects were closed directly or covered by paraumbilical island flaps.

RESULTSThe 17 flap size ranged from 5 cm x 3 cm to 19 cm x 14 cm. 16 flaps survived completely except 1 flap with partial necrosis, which was closed by free skin graft. The donor site defects were closed directly in 10 cases, and covered by paraumbilical island flaps in 7 flaps without no flap necrosis. The abdomen had a good appearance.

CONCLUSIONSGood appearance can be achieved with free superficial iliac circumflex artery skin flaps for the defects on face, neck, foot, hand, ankle and lower leg. Paraumbilical island flap can be used for the donor site defects.

Arteries ; Foot ; Free Tissue Flaps ; blood supply ; transplantation ; Humans ; Reconstructive Surgical Procedures ; Skin ; Skin Transplantation ; Transplant Donor Site ; surgery ; Wounds and Injuries ; surgery