Application and Prospect of RNA Profiling Analysis in Forensic Body Fluid Identification / 法医学杂志

In forensic physical evidence identification, the accurate identification of the individual origin and their body fluid composition of the biological samples obtained from the crime scene play a critical role in determining the nature of a crime. In recent years, RNA profiling has become one of the fastest developing methods for body fluids identification. Due to the characteristics of tissue or body fluid specific expression, various types of RNA markers have been proven to be promising candidate markers for body fluids identification in previous studies. This review summarizes the research progress of RNA markers in body fluids identification, including the RNA markers that have been effectively verified in current research and their advantages and disadvantages. Meanwhile, this review prospects the application of RNA markers in forensic medicine.

Molecular characterization of Staphylococcus aureus ST6 and ST7 isolates from food-borne illness outbreaks / 中华预防医学杂志

Objective: To analyze the Staphylococcal enterotoxins, Staphylococcal enterotoxin genes, drug resistance and molecular typing of 41 Staphylococcus aureus isolates from 2 food-borne illness outbreaks on 21 August and 27 September 2020 in Guangzhou. Methods: A total of 41 Staphylococcus aureus isolates from 2 outbreaks were analyzed by multilocus sequence typing (MLST) and spa typing. The Staphylococcal enterotoxins typing and the Staphylococcal enterotoxin genes of the isolates were analyzed by ELISA and PCR, respectively. The antimicrobial susceptibility of the isolates was performed by disc diffusion. 21 Staphylococcus aureus isolates were characterized using whole genome sequencing (WGS). Based on the whole genome single nucleotide polymorphism (SNP), the phylogenetic tree was constructed by Snippy. Results: 41 Staphylococcus aureus isolates were divided into 2 types by MLST and spa typing: ST6-t701 and ST7-t091. 2 ST7-t091 isolates were identified as methicillin-resistant Staphylococcus aureus (MRSA). 25 ST7-t091 isolates and 14 ST6-t701 isolates were methicillin-sensitive Staphylococcus aureus (MSSA), and were resistant to 7 and 6 antibiotics, respectively. All isolates were positive for sea by PCR. WGS revealed all 21 isolates carried scn, sak, sea, hla, hld, hlgA, hlgB, hlgC, lukD virulence genes. The results showed the isolates contained an immune evasion cluster type D which located in bacteriophage ϕSa3. The SNP phylogenetic tree showed 2 MRSA ST7-t091 were constituted a separate clade from the 12 MSSA ST7-t091 isolates and 7 ST6-t701 isolates showed high similarity to each other. Conclusion: Base on the results of phylogenetic analysis, the 2 food-borne illness outbreaks occurred on 21 August and 27 September 2020 are caused by the combination of the MRSA ST7-t091 strain and the MSSA ST7-t091 strain, and the MSSA ST6-t701 strain, respectively. All isolates have high level of antibiotic resistance and carry high virulent genes.

Xiaojindan Extract Modulated Macrophage Polarization by Targeting PI3K/Akt Pathway / 中国实验方剂学杂志

ObjectiveTo explore the effect and mechanism of Xiaojindan extract （XJD） on macrophage polarization. MethodLipopolysaccharide （LPS） and interleukin-4 （IL-4） were used to induce M1 and M2 polarization of RAW264.7 cells. The influence of 10-80 mg·L-1 XJD on cell proliferation was detected by Cell Counting Kit-8 （CCK-8） assay. Nitric oxide （NO） and interleukin-6 （IL-6） release was explored by Griess assay and enzyme-linked immunosorbent assay （ELISA）， respectively. The mRNA expression of M1 and M2 macrophage markers was measured by real-time quantitative polymerase chain reaction （Real-time PCR）， and the CD206+ expression was determined by flow cytometry. The activation of phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） pathway was analyzed by western blot. Result10-80 mg·L-1 XJD showed no marked cytotoxicity in LPS （0.5 mg·L-1）- or IL-4 （20 μg·L-1）-induced RAW264.7 cells. Compared with the control group， LPS significantly promoted the expression of M1 macrophage markers （P<0.01）， including increased NO and IL-6 release （P<0.01） and upregulated mRNA expression of interleukin-1β （IL-1β）， inducible nitric oxide synthase （iNOS）， cyclooxygenase-2 （COX-2） and tumor necrosis factor-α （TNF-α） （P<0.01）. Compared with LPS-induced group， 20-80 mg·L-1 XJD decreased the release of NO and IL-6 in a dose-dependent manner （P<0.01）， and similarly 10-80 mg·L-1 XJD suppressed the mRNA expression of IL-1β， iNOS， COX-2 and TNF-α （P<0.01）. Compared with the control group， IL-4 obviously increased the expression of M2 macrophage markers （P<0.01）， including increased CD206+ cell population and upregulated mRNA expression of arginine-1 （Arg-1）， interleukin-10 （IL-10）， interleukin-13 （IL-13） and transforming growth factor-β1 （TGF-β1）. Compared with IL-4-induced group， 10-80 mg·L-1 XJD dose-dependently decreased CD206+ cell population （P<0.01） and inhibited the mRNA expression of Arg-1， IL-10， IL-13 and TGF-β1 （P<0.01）. Western blot showed that XJD significantly downregulated the activation of PI3K/Akt pathway as compared to LPS- and IL-4-induced groups （P<0.05， P<0.01）. ConclusionXJD significantly inhibited the macrophage polarization in the LPS- and IL-4-induced RAW264.7 cells by targeting PI3K/Akt pathway.

Study on regulation of CYP450 enzyme system to reduce liver toxicity through compatibility of Aconiti Kusnezoffii Radix Cocta with Chebulae Fructus and Glycyrrhizae Radix et Rhizoma / 中国中药杂志

Aconiti Kusnezoffii Radix Cocta is one of the most commonly used medicinal materials in Mongolian medicine. Due to the strong toxicity of Aconiti Kusnezoffii Radix Cocta, Mongolian medicine often uses Chebulae Fructus, Glycyrrhizae Radix et Rhizoma to reduce the toxicity, so as to ensure the curative effect of Aconiti Kusnezoffii Radix Cocta while ensuring its clinical curative effect, but the mechanism is not clear. The aim of this study was to investigate the effects of Chebulae Fructus, Glycyrrhizae Radix et Rhizoma and Aconiti Kusnezoffii Radix Cocta on the mRNA transcription and protein translation of cytochrome P450(CYP450) in the liver of normal rats. Male SD rats were randomly divided into negative control(NC) group, phenobarbital(PB) group(0.08 g·kg~(-1)·d~(-1)), Chebulae Fructus group(0.254 2 g·kg~(-1)·d~(-1)), Glycyrrhizae Radix et Rhizoma group(0.254 2 g·kg~(-1)·d~(-1)), Aconiti Kusnezoffii Radix Cocta group(0.254 2 g·kg~(-1)·d~(-1))and compatibility group(0.254 2 g·kg~(-1)·d~(-1),taking Aconiti Kusnezoffii Radix Cocta as the standard). After continuous administration for 8 days, the activities of total bile acid(TBA), alkaline phosphatase(ALP), amino-transferase(ALT) and aspartate aminotransferase(AST)in serum were detected, the pathological changes of liver tissue were observed, and the mRNA and protein expression levels of CYP1 A2, CYP2 C11 and CYP3 A1 were observed. Compared with the NC group, the serum ALP, ALT and AST activities in the Aconiti Kusnezoffii Radix Cocta group were significantly increased, and the ALP, ALT and AST activities were decreased after compatibility. At the same time, compatibility could reduce the liver injury caused by Aconiti Kusnezoffii Radix Cocta. The results showed that Aconiti Kusnezoffii Radix Cocta could inhibit the expression of CYP1 A2, CYP2 C11 and CYP3 A1, and could up-regulate the expression of CYP1 A2, CYP2 C11 and CYP3 A1 when combined with Chebulae Fructus and Glycyrrhizae Radix et Rhizoma. The level of translation was consistent with that of transcription. The compatibility of Chebulae Fructus and Glycyrrhizae Radix et Rhizoma with Aconiti Kusnezoffii Radix Cocta could up-regulate the expression of CYP450 enzyme, reduce the accumulation time of aconitine in vivo, and play a role in reducing toxicity, and this effect may start from gene transcription.

Edible Bird's Nest Attenuates Menopause-Related Bone Degeneration in Rats via Increaing Bone Estrogen-Receptor Expression / 中国结合医学杂志

OBJECTIVE@#To investigate the mechanistic basis for the attenuation of bone degeneration by edible bird's nest (EBN) in ovariectomized rats.@*METHODS@#Forty-two female Sprage-Dawley rats were randomized into 7 groups (6 in each group). The ovariectomized (OVX) and OVX + 6%, 3%, and 1.5% EBN and OVX +estrogen groups were given standard rat chow alone, standard rat chow +6%, 3%, and 1.5% EBN, or standard rat chow +estrogen therapy (0.2mg/kg per day), respectively. The sham-operation group was surgically opened without removing the ovaries. The control group did not have any surgical intervention. After 12 weeks of intervention, blood samples were taken for serum estrogen, osteocalcin, and osteoprotegerin, as well as the measurement of magnesium, calcium abd zinc concentrations. While femurs were removed from the surrounding muscles to measure bone mass density using the X-ray edge detection technique, then collected for histology and estrogen receptor (ER) immunohistochemistry.@*RESULTS@#Ovariectomy altered serum estrogen levels resulting in increased food intake and weight gain, while estrogen and EBN supplementation attenuated these changes. Ovariectomy also reduced bone ER expression and density, and the production of osteopcalcin and osteorotegerin, which are important pro-osteoplastic hormones that promote bone mineraliztion and density. Conversely, estrogen and EBN increased serum estrogen levels leading to increased bone ER expression, pro-osteoplastic hormone production and bone density (all P<0.05).@*CONCLUSION@#EBN could be used as a safe alternative to hormone replacement therapys for managing menopausal complications like bone degeneration.

Correlation analysis of the degree of demodex infection with corneal cell density and ocular surface function in BKC patients / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To investigate the correlation between demodex infection with corneal cell density changes and ocular surface function in patients with blepharo kerato conjunctivitis(BKC). <p>METHODS: Ninety-four patients with BKC(BKC group)at Department of Ophthalmology of our hospital from July 2019 to July 2020 were selected as the research objects, in addition, 80 matched healthy volunteers were selected as control group. The BKC patients were divided into infected group(45 cases)and uninfected group(49 cases)according to whether they were infected with demodex. According to the number of demodex detected in eyelashes, there were 17 cases of suspicious infection, 18 cases of moderate infection and 10 cases of severe infection. All subjects were examined by laser confocal microscopy, and the cell density in the superficial stromal layer of the central cornea and peripheral cornea was calculated. The ocular surface function \〖Schirmer test, Ocular Surface Disease Index(OSDI)\〗, eyelid margin abnormality score, corneal fluorescence stain and tear film break-up time(TF-BUT)of patients with BKC were examined, and the correlation between demodex infection with corneal cell density and ocular surface function in patients with BKC was analyzed. <p>RESULTS: Compared with those in the control group, the cell density in the superficial stromal layer of the central cornea and peripheral cornea was lower in the BKC group(<i>P</i><0.05), and the OSDI, eyelid margin abnormality score and corneal fluorescence stain score were higher(<i>P</i><0.05); the cell density in the superficial stromal layer of the central cornea and peripheral cornea of patients in uninfected group, patients with suspicious demodex infection, moderate demodex infection and severe demodex infection decreased in turn(<i>P</i><0.05), and the OSDI, eyelid margin abnormality score and corneal fluorescence stain score increased significantly in turn(<i>P</i><0.05); the degree of demodex infection was negatively correlated with the cell density in the superficial stromal layer of the central cornea and peripheral cornea in patients with BKC(<i>P</i><0.05), and was positively correlated with OSDI, eyelid margin abnormality score and corneal fluorescence stain score(<i>P</i><0.05).<p>CONCLUSION: The severity of demodex infection has a significant negative correlation with the cell density in the superficial stromal layer of the central cornea and peripheral cornea in patients with BKC, has a significant positive correlation in patients with ocular surface dysfunction.

Correlation analysis of the degree of demodex infection with corneal cell density and ocular surface function in BKC patients / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To investigate the correlation between demodex infection with corneal cell density changes and ocular surface function in patients with blepharo kerato conjunctivitis(BKC). <p>METHODS: Ninety-four patients with BKC(BKC group)at Department of Ophthalmology of our hospital from July 2019 to July 2020 were selected as the research objects, in addition, 80 matched healthy volunteers were selected as control group. The BKC patients were divided into infected group(45 cases)and uninfected group(49 cases)according to whether they were infected with demodex. According to the number of demodex detected in eyelashes, there were 17 cases of suspicious infection, 18 cases of moderate infection and 10 cases of severe infection. All subjects were examined by laser confocal microscopy, and the cell density in the superficial stromal layer of the central cornea and peripheral cornea was calculated. The ocular surface function \〖Schirmer test, Ocular Surface Disease Index(OSDI)\〗, eyelid margin abnormality score, corneal fluorescence stain and tear film break-up time(TF-BUT)of patients with BKC were examined, and the correlation between demodex infection with corneal cell density and ocular surface function in patients with BKC was analyzed. <p>RESULTS: Compared with those in the control group, the cell density in the superficial stromal layer of the central cornea and peripheral cornea was lower in the BKC group(<i>P</i><0.05), and the OSDI, eyelid margin abnormality score and corneal fluorescence stain score were higher(<i>P</i><0.05); the cell density in the superficial stromal layer of the central cornea and peripheral cornea of patients in uninfected group, patients with suspicious demodex infection, moderate demodex infection and severe demodex infection decreased in turn(<i>P</i><0.05), and the OSDI, eyelid margin abnormality score and corneal fluorescence stain score increased significantly in turn(<i>P</i><0.05); the degree of demodex infection was negatively correlated with the cell density in the superficial stromal layer of the central cornea and peripheral cornea in patients with BKC(<i>P</i><0.05), and was positively correlated with OSDI, eyelid margin abnormality score and corneal fluorescence stain score(<i>P</i><0.05).<p>CONCLUSION: The severity of demodex infection has a significant negative correlation with the cell density in the superficial stromal layer of the central cornea and peripheral cornea in patients with BKC, has a significant positive correlation in patients with ocular surface dysfunction.

Lipopolysaccharide inhibits lipophagy in HepG2 cells via activating mTOR pathway / 生理学报

This study aimed to investigate the effect of lipopolysaccharide (LPS) on lipophagy in hepatocytes and the underlying mechanism. Human hepatoma cell line HepG2 was cultured in vitro, treated with 0.1 mmol/L palmitic acid (PA), and then divided into control group (0 μg/mL LPS), LPS group (10 μg/mL LPS), LPS+DMSO group and LPS+RAPA (rapamycin, 10 μmol/L) group. Lipid accumulation in hepatocytes was observed by oil red O staining. The autophagic flux of the cells was assessed using confocal laser scanning microscope after being transfected with autophagy double-labeled adenovirus (mRFP-GFP-LC3). The level of intracellular lipophagy was visualized by the colocalization of lipid droplets (BODIPY 493/503 staining) and lysosomes (lysosome marker, lysosomal associated membrane protein 1, LAMP1). The expression levels of mammalian target of rapamycin (mTOR), phosphorylated mTOR (p-mTOR), ribosome protein subunit 6 kinase 1 (S6K1), p-S6K1, LC3II/I and P62 protein were examined by Western blot. The results showed that the number of red lipid droplets stained with oil red O was significantly increased in LPS group compared with that in control group (P < 0.001). Moreover, in LPS group, the number of autophagosomes was increased, while the number of autophagolysosomes and the colocalization rate of LAMP1 and BODIPY were significantly decreased (P < 0.05). Meanwhile, the ratios of p-mTOR/mTOR and p-S6K1/S6K1, the ratio of LC3II/LC3I and the protein expression of P62 were significantly increased (P < 0.05) in LPS group. Furthermore, compared with LPS+DMSO group, RAPA treatment obviously reduced the number of lipid droplets and autophagosomes, and raised the number of autophagolysosomes and the colocalization rate of LAMP1 and BODIPY (P < 0.05). In conclusion, the results demonstrate that LPS inhibits lipophagy in HepG2 cells via activating mTOR signaling pathway, thereby aggravating intracellular lipid accumulation.

25-hydroxyvitamin D, bone source alkaline phosphatase and ultrasound bone density combined in detection of rickets in infants / 上海预防医学

Objective To analyze the application of serum 25-hydroxyvitamin D (25-OH-D), bone source alkaline phosphatase (BAP) and ultrasonic bone mineral density (BMD) in the detection of infantile rickets aged 3-12 months. Methods Six to 12 months old rickets infants and healthy ones were randomly selected from March to December 2018 in a hospital in Changning, who were divided into two groups as observation or control group (160 infants in each).Two groups were respectively tested with serum 25-OH-D, BAP activity and ultrasonic bone density, so as to explore the diagnostic efficacy of the three combined tests for infant rickets. Results The serum 25-OH-D level and the BMD in the observation group were significantly lower than that in the control group (P < 0.05), and the abnormal detection rate of BAP in the control group was significantly lower than that in the observation group (P < 0.05).There was no statistical significance (P>0.05) between the three detection methods, but the sensitivity and accuracy of the three detection methods combined were significantly better than that of any single detection method (P < 0.05). Conclusion The combined detection with serum 25-OH-D level, BAP activity and ultrasonic BMD can significantly improve the detection efficiency of rickets in infants aged 6-12 months, which is valuable and worthy of clinical application.

Effect of Tetrandrine on Col-I and FN in TGF-β1-induced MRC-5 Cells / 中国实验方剂学杂志

Objective:To investigate the effect of tetrandrine on transforming growth factor-β1（TGF-β1）stimulated MRC-5 cells. Method:Different concentrations of TGF-β1 (0, 2.5, 5, 10, 20, 40 μg·L-1) were applied to MRC-5 cells. Proliferation toxicity of TGF-β1 to MRC-5 was detected by cell counting kit-8 (CCK-8) method. Detection of alpha smooth muscle actin (α-SMA) and Vimentin's expression levels in MRC-5 by Western blot. Detection of changes of collagen I(Col-I) and fibronectin (FN)'s expression levels in MRC-5 supernatants by enzyme linked immunosorbent assay(ELISA) kit. And the appropriate concentration of TGF-β1 activated MRC-5 cells was screened. The appropriate concentration of TGF-β1 and different concentrations of Tet (0, 2.5, 5, 10, 20, 40 μmol·L-1) were applied to MRC-5 cells, and CCK-8 method was used to screen safe concentration again. Western blot was used to detect changes in α-SMA and Vimentin expression levels in MRC-5 cells, and ELISA method to detect changes in Col-I and FN in MRC-5 cell supernatant. Result:Compared with the blank group, 20，40 μg·L-1 of TGF-β1 had toxic effects on MRC-5 cells at 24 hours (P<0.05), and 10，20，40 μg·L-1 of TGF-β1 had toxic effects on MRC-5 cells at 48 h (P<0.05).When Tet is added for 24 h, the half inhibitory concentration (IC50) value was 14.07 μmol·L-1, and when cultured for 48 h, the IC50 value was 7.51 μmol·L-1. Compared with the blank group, the relative contents of α-SMA, FN and Col-I in the 5 μg·L-1 of TGF-β1 group were obviously increased (P<0.05), and the relative contents of Vimentin were significantly increased (P<0.01), and the relative contents of FN and Col-I, α-SMA and Vimentin in 10 μg·L-1 group were significantly increased (P<0.01). 10 μg·L-1 of TGF-β1 was co-cultured with Tet at different concentrations. Compared with the TGF-β1 group, the relative levels of α-SMA, Vimentin and FN in the 5 μmol·L-1 of Tet group were significantly reduced (P<0.01), and the relative levels of Col-I were obviously reduced (P<0.05). In the Tet 10 μmol·L-1 group, the relative contents of the α-SMA, Vimentin, FN and Col-I were significantly reduced (P<0.01). Conclusion:TGF-β1 can increase the levels of Col-I, FN and other extracellular matrices in MRC-5 cells, and Tet can effectively inhibit the occurrence of this change. It is suggested that Tet may inhibit secreting extracellular matrix of fibroblasts in the formation of pulmonary fibrosis.

Effect of Effective Components of Salvia Miltiorrhiza on TGF-β1 Induced Interstitial Transformation of Pulmonary Epithelial Cells / 中国实验方剂学杂志

Objective::To screen out the effective components of Salvia miltiorrhiza by establishing an in vitro model of pulmonary epithelial mesenchymal transformation. Method::Different concentrations of salvianolic acid A (10, 20, 40, 80, 160 μmol·L-1), salvianolic acid B (10, 20, 40, 80, 160 μmol·L-1), tanshinol (10, 20, 40, 80, 160 μmol·L-1), tanshinoneⅡA (10, 20, 40, 80, 160 μmol·L-1) and the blank group were applied to A549 cell, cell proliferation and cytotoxicity assay (MTS) were used to detect the proliferation effect of menthol on A549 cells.After screening the safe concentration of the active ingredients of salvia miltiorrhiza by MTS, cells were divided into blank group, model group, salvianolic acid A group, salvianolic acid B group, tanshinol group and tanshinoneⅡA.Then, the inhibitory effect of the active ingredients of salvia miltiorrhiza on the proliferation of A549 cells induced by TGF-β1 was detected by MTS. Enzyme linked immunosorbent assay (ELISA) method to detect salvia miltiorrhiza effective component of fiber protein(FN), collagen type I (COL-Ⅰ) expression. Based on the above results, the active components of salvia miltiorrhiza, which have best inhibition were screened out, and their effects on the expression of E-calcium-viscosity (E-Cad) protein were detected by Western blot. Result::Compared with blank group, salvianolic acid A 40 μmol·L-1, salvianolic acid B 160 μmol·L-1, tanshinol 160 μmol·L-1 had toxic effects on A549 cells (P<0.05). In the non-toxic concentration range, compared with the model group, salvianolic acid A 10, 20 μmol·L-1, salvianolic acid B 80 μmol·L-1 showed inhibition effect after 24 h culture (P<0.05). After 72 h culture, salvianolic acid A 5, 10, 20 μmol·L-1, salvianolic acid B 40, 80 μmol·L-1inhibition effect was very significant (P<0.01). ELISA results showed that with the blank group, model group cells the expression of FN and COL-Ⅰ increased significantly (P < 0.01). Compare with model group, salvianolic acid A 20 μmol·L-1, salvianolic acid B 80 μmol·L-1 inhibited FN and COL-Ⅰ(P<0.05). Western blot results showed that salicylic acid A and salicylic acid B had protective effects on E-Cad (P<0.01). Conclusion::Salvianolic acid A and salvianolic acid B have inhibitory effects on epithelial mesenchymal transformation by TGF-β1, which may be the main effective components of salvianolic acid in the treatment of pulmonary fibrosis.

CT imaging and quantitative analysis in the differential diagnosis of hepatic epithelioid hemangioendothelioma and hepatic metastasis / 中华肝胆外科杂志

Objective To study the use of abdominal enhanced CT imaging and quantitative index analysis in the differential diagnosis of hepatic epithelioid hemangioendothelioma (HEH) and hepatic metastasis.Methods A study group of 12 patients with HEH who underwent abdominal enhanced CT scanning at the First Affiliated Hospital of Zhengzhou University from February 2014 to October 2018 was retrospectively compared with a control group of 52 patients with hepatic metastases diagnosed clinically and by imaging examinations.The general information and imaging data of these patients were collected and analyzed.Results The lesions in the 2 groups mainly presented as multiple and diffuse lesions.The diffuse lesions of HEH often fused into strips.The hepatic metastasis group showed a higher CT attenuation and TNR in the portal vein phase than the HEH group (P ＜ 0.05).The area under the ROC curves of the two indexes were 0.756 and 0.841 respectively.The centers of the lesions showed almost no or slightly homogeneous enhancement in the HEH group,while the liver metastasis group showed slightly and moderately heterogeneous enhancement,with a significant difference between the two groups (P ＜ 0.05).Female,subcapsular distribution,capsular contraction,target ring sign and lollipop sign were independent risk factors for HEH (P ＜0.05),while a high CT attenuation and TNR in the portal vein phase,elevated tumor markers and lymph node metastasis were independent risk factors for liver metastasis on logistic regression analysis (P ＜ 0.05).Conclusions CT attenuation,TNR,central enhancement features in the portal vein phase,special signs and secondary changes of lesions were helpful for the differential diagnosis between HEH and liver metastasis.

Application Prospect of Massively Parallel Sequencing in Mixed Stain Detection / 法医学杂志

Mixed stains is the common biological sample in sexual crime cases. Its analysis and DNA profiles interpretation are one of the difficulties in forensic examination. The current genetic marking of mixed stain detection mainly rely on capillary electrophoresis （CE） separation technology, and the analysis methods of the results are mainly inclusion rate and likelihood methods. Because CE has limited resolution and is not able to exploit the efficacy of each genetic marker, its ability to split mixed stain is limited. In recent years, the emerging massively parallel sequencing （MPS） technique not only can obtain the base sequence information of genetic markers, but also is capable of detecting multiple genetic markers simultaneously, and thus derives new analytical methods, bringing new opportunities for forensic detection and analysis of mixed stain. This paper intends to review the application prospects of conventional mixed stain analyses and MPS technique, therefore to provide references for later research and practice.

The application value of spectral scanning combined with iterative reconstruction in double-low technique scanning for severe liver cirrhosis / 中华放射医学与防护杂志

Objective To investigate the clinical feasibility of spectral monochromatic imaging combined with adaptive statistical iterative reconstruction-V (ASIR-V) in upper abdominal enhanced scanning with double-low technique for severe liver cirrhosis.Methods Totally 126 cases performed abdominal contrast-enhanced CT scanning were collected prospectively and assigned to 3 groups with 42 cases in each group with random number table method.The filtered back projection algarithm,120 kV and contrast agent dose of 420 mgI/kg were used for the cases of control group.The cases for spectral group and combined group were scanned with spectral imaging mode and contrast agent dose of 300 mgI/kg.The 60 keV monochromatic images combined with pre-0％ and post-40％ ASIR-V were selected and analyzed for spectral group.Combined group with pre-40％ ASIR-V was divided into 2 subgroups with 50 keV,post -50％ ASIR-V and 60 keV,post-40％ ASIR-V separately.One-way ANOVA test was used for analysis of radiation dose and quantitative parameters,and Rank sum test was used for image subject evaluation.Results The CT numbers and CNR had significant differences among spectral group,combined group and control group(F=4.293-13.134,P＜0.05) except for the images of liver parenchyma in PVP and that of 50 keV combined 50％ASIR-V group were higher than that of control group (q=1.825-3.736,P＜0.05).No significant differences existed for image noise and overall image quality scores of organs in four groups.The visualization of hepatic vascular branches in 50 keV combined 50％ASIR-V group was higher than that of other three groups(Z=2.793-6.328,P＜0.05).The radiation dose of combined group was lower than that of spectral and control group (q =-4.879--2.531,P ＜ 0.001).Conclusions Spectral monochromatic imaging combined with pre-and post-ASIR-V can reduce contrast agent dose and radiation dose without degrading image quality for the severe liver cirrhosis in upper abdominal enhanced scanning.

Pharmacodynamic Evaluation of Tanreqing Inhalation Solution / 中国实验方剂学杂志

Objective: To clarify the antitussive, expectorant, antipyretic and anti-inflammatory effects of Tanreqing inhalation solution, and provide basis and data support for further research and development of this preparation. Method: The methods of cough induced by ammonia and tracheal phenol red excretion were used to observe the antitussive and expectorant effects of Tanreqing inhalation solution in mice. The fever model of rats was established by intraperitoneal injection of bacterial lipopolysaccharide(LPS) to observe the antipyretic effect of the Tanreqing inhalation solution, the acute pneumonia model of rats was established by atomizing LPS inhalation, and the anti-inflammatory effect of Tanreqing inhalation solution was observed. Result: Tanreqing inhalation solution could reduce the number of coughs in mice induced by ammonia water, increase the amount of phenol red excretion in mouse trachea, decrease the levels of body temperature and its related regulatory factors of prostaglandin E2(PGE2) and cyclic adenosine monophosphate(cAMP) of rats induced by LPS, decrease the white blood cell(WBC) count and the neutrophil ratio(NEUT) in bronchoalveolar lavage fluid(BALF) of rats with LPS-induced acute pneumonia, and reduce the levels of nuclear transcription factor-κB(NF-κB) and interleukin-1β(IL-1β) in lung tissue. Conclusion: Tanreqing inhalation solution has obvious antitussive, expectorant, antipyretic and anti-inflammatory effects, which is worthy of further development and promotion.

Effect of Different Administration Methods with Bleomycin on Pulmonary Fibrosis in Rats / 中国实验方剂学杂志

Objective:Compare the effects of 3 administration methods (tracheal perfusion, tail vein injection and aerosol inhalation) with bleomycin (BLM) in inducing pulmonary fibrosis in rats, in order to find out the optimal administration methods. Method:Eighty sprague-dawley (SD) male rats with SPF were randomly divided into aerosol inhalation blank group, single tracheal perfusion group(10 mg·kg-1), multiple tracheal perfusion group(5 mg·kg-1), single intravenous injection group(150 mg·kg-1), multiple intravenous injection group(50 mg·kg-1), single aerosol inhalation group (30 min)and multiple aerosol inhalation group(30 min). The mortality and body weight of rats in each group were observed at 7 d, 14 d and 28 d after the administration. And 28 days later after the administration, the lung coefficients of rats in each group were observed, paraffin sections were prepared, hematoxylin-eosin staining (HE) and Masson staining were performed, and the contents of hydroxyproline (HYP) and plasminogen activator inhibitor-1 (PAI-1) in lung tissues were detected by enzyme-linked immunosorbent assay (ELISA), so as to evaluate the alveoli inflammation and pulmonary fibrosis of rats in each group. Result:Compared with the aerosol inhalation blank group, the rats in the trachea perfusion group had the highest mortality among the drug treatment groups. The pulmonary coefficients of rats in the multiple intravenous injection group and the multiple inhalation group were significantly higher than those in the blank group(PPPConclusion:Bleomycin was inhaled repeatedly to establish pulmonary fibrosis model. The pathological injury and physiological indexes of the model rats were relatively stable, which conforms with the evolution process of pulmonary fibrosis.

Mechanism of Dichroa Alkali Salt Inducing Vomiting Based on Pica in Mice / 中国实验方剂学杂志

Objective: To study the possible mechanism of dichroa alkali salt (DAS) in inducing vomiting. Method: Mice pica model was used to observe the antagonistic effect of the three different kinds of antiemetic drugs[dopamine receptor antagonist metoclopramide, 5-hydroxytryptamine 3 (5-HT3) receptor antagonist ondansetron and neurokinin-1 receptor antagonist aprepitant] on body mass, food intake, kaolin consumption, diarrhea and death induced by DAS to preliminarily clarify the possible pathogenic pathway of DAS. Then, the expression of 5-HT and substance P(SP) in ileum and medulla of mice induced by DAS alone at different time points was detected by enzyme-linked immunosorbent assay to confirm whether DAS could affect the changes of these two neurotransmitters. Result: After treatment with ondansetron and aprepitant, DAS-induced reduction in food intake of mice was significantly improved on the 4th day after continuous administration and on the 1st day after drug administration (Prd day after administration, DAS-induced body mass loss of mice was significantly improved (PConclusion: The mice pica model can be used to effectively characterize DAS-induced vomiting. DAS-induced pica in mice may be associated with the increase of 5-HT and SP in ileum and medulla. Ondansetron and aprepitant can effectively antagonize DAS-induced pica in mice.

Investigation of Physicochemical Properties and Determination Methods of Liposomal Entrapment Efficiency of Dichroa Alkali Hydrochloride / 中国实验方剂学杂志

Objective: To study on the physical and chemical properties of dichroa alkali hydrochloride and to establish a method for the determination of entrapment efficiency of dichroa alkali hydrochloride liposomes. Method: HPLC was used to determine the content of dichroa alkali hydrochloride with mobile phase of acetonitrile-water-triethylamine-glacial acetic acid(9:91:0.35:0.75) and detection wavelength at 265 nm.The oil-water partition coefficient of this compound in different pH range was measured by shake flask method.The stability of the dichroa alkali hydrochloride in phosphate buffer solution with different pH after sterilization at 125℃ for 30 min was investigated.Ammonium sulfate gradient method was used to prepare dichroa alkali hydrochloride liposomes.The microcolumn was prepared by dextran gel and cation exchange resin,respectively.Then the free drug and liposome were separated by centrifugation,the drug content was measured,and the encapsulation efficiency was calculated.The t-test was performed using SPSS 20.0 software,the differences between these two methods were compared. Result: In the pH 6-9,the oil-water partition coefficient of dichroa alkali hydrochloride increased with increasing of pH,which was between 0.016 and 1.44;the recovery rate of dichroa alkali hydrochloride after sterilization was 37.16%-57.91%.Between the dextran gel microcolumn centrifugation and the cation exchange resin microcolumn centrifugation,there was no significant difference in the entrapment efficiency of the liposomes. Conclusion: Dichroa alkali hydrochloride is suitable for preparation of liposomes.However,its stability is not ideal,so the experimental temperature should be strictly controlled in the preparation process.Dextran gel microcolumn centrifugation and cation exchange resin microcolumn centrifugation can be used to determine the entrapment efficiency of dichroa alkali hydrochloride liposomes,and the cation exchange resin microcolumn centrifugation is suggested after comparison.

Protective Effect of Renshen Sinitang and Its Active Ingredients on Myocardial Cell Injury Induced by Pentobarbital Sodium / 中国实验方剂学杂志

Objective: To explore the protective effect and mechanisms of Renshen Sinitang and its active ingredients on cardiomyocyte injury induced by pentobarbital sodium. Method: H9C2 cells were sub-cultured with ginsenoside Rb2 0.01, 0.1, 1 μmol ·L-1, Re 0.01, 0.1, 1 μmol·L-1, isoliquiritigenin 20, 40, 80 μmol·L-1, glycyrrhetinic acid 10, 20, 40 μmol·L-1, Renshen Sinitang, 10, 100, 400 mg·L-1, for 4 h. After treatment with 0.1% of sodium pentobarbital for 30 min, cell viability, lactate dehydrogenase (LDH), lipid peroxide malondialdehyde (MDA), Na+-K+-adenosine triphosphate(ATP) ase, Ca2+-ATPase activity, and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) were used to detect the expressions of peroxisome proliferative activated receptor-1α (PGC-1α), B-cell lymphoma-2 associated X protein(Bax) and cysteine aspartate-specific protease-3(Caspase-3) mRNA. Result: Renshen Sinitang and its active ingredients have a protective effect on heart failure cell model. Compared with the normal group, the cell survival rate of the model group decreased significantly, while the LDH and MDA contents increased significantly, and the Na+-K+-ATPase activity increased. Ca2+-ATPase activity was significantly decreased, PGC-1α mRNA expression was down-regulated, Bax and Caspase-3 mRNA expressions indicates the modeling(P+-K+-ATPase activity, increased Ca2+-ATPase activity, up-regulated PGC-1α mRNA expression, and inhibited Bax and Caspase-3 mRNA expression (PPConclusion: Renshen Sinitang and its active ingredients have a significant protective effect on heart failure cell model, and its mechanisms of action are related to anti-oxidation, improvement of mitochondrial energy metabolism and inhibition of mitochondrial apoptosis pathway.

Trans-ethnic analysis of susceptibility variants in IgA nephropathy / 北京大学学报(医学版)

OBJECTIVE@#To compare the genetic architecture of susceptibility variants of IgA nephropathy (IgAN) in Chinese and Europeans.@*METHODS@#We selected the independent genome-wide significant variants of IgAN in European population as candidate variants. Their associations, risk alleles, risk allele frequencies, odds ratios and population attributable risk scores were derived and calculated, then compared with those in the current Chinese population, including 1 194 IgAN patients and 902 controls. Using the significant variants, genetic risk scores were calculated and compared between the East Asians and the Europeans. The correlation between the genetic risk scores and clinical manifestations was also evaluated.@*RESULTS@#There were 16 independent single nucleotide polymorphisms (SNPs) located in 11 loci showing significantly association with susceptibility to IgAN in the Europeans. 93.75% (15/16) of them also showed significant associations in the Chinese (P<0.05). The effects of all the associated SNPs were in the same direction, either risk or being protective for IgAN, between the Chinese and the Europeans. On the contrary, remarkable higher risk allelic odds ratio (P=1.94×10-2), higher risk allele frequency (P=3.09×10-2), and higher population attributable risk (P=3.03×10-4) were observed for most of the associated SNPs in the Chinese than in the Europeans. Furthermore, genetic risk scores were significantly larger in the Asian populations compared with the Europeans (P=1.78×10-163). While there was no significance among the subpopulations in both the East Asians and the Europeans. Compared with the healthy controls, the genetic risk score in the IgAN patients was significantly larger (P=3.60×10-27). Clinical analysis showed the genetic risk score was positively associated with serum levels of IgA and IgA1, phases of chronic kidney disease and Haas grades.@*CONCLUSION@#Our study provides further evidence in the shared genetic architecture between Chinese and Europeans, while differences with respect to the effect sizes and risk allele frequencies across ethnicities, contributing partially to the differences of disease prevalence.