Enzymatic Digestion Method Coupled with Artificial Intelligence Techniques in Forensic Drowning Diatom Detection / 中山大学学报(医学科学版)

ObjectiveArtificial intelligence （AI） full smear automated diatom detection technology can perform forensic pathology drowning diatom detection more quickly and efficiently than human experts.However， this technique was only used in conjunction with the strong acid digestion method， which has a low extraction rate of diatoms. In this study， we propose to use the more efficient proteinase K tissue digestion method （hereinafter referred to as enzyme digestion method） as a diatom extraction method to investigate the generalization ability and feasibility of this technique in other diatom extraction methods. MethodsLung tissues from 6 drowned cadavers were collected for proteinase K ablation and made into smears， and the smears were digitized using the digital image matrix cutting method and a diatom and background database was established accordingly.The data set was divided into training set， validation set and test set in the ratio of 3：1：1， and the convolutional neural network （CNN） models were trained， internally validated， and externally tested on the basis of ImageNet pre-training. ResultsThe results showed that the accuracy rate of the external test of the best model was 97.65 %， and the area where the model features were extracted was the area where the diatoms were located. The best CNN model in practice had a precision of more than 80 % for diatom detection of drowned corpses. ConclusionIt is shown that the AI automated diatom detection technique based on CNN model and enzymatic digestion method in combination can efficiently identify diatoms and can be used as an auxiliary method for diatom detection in drowning identification.

Preparation of drug loading system of black phosphorus nanosheets and its anti-ischemic brain injury / 药学学报

In this study, black phosphorus nanosheets (BP) were prepared by the ordinary liquid phase method, and resveratrol was loaded on the BP after being modified by polyethylene glycol. The brain targeting of BP was investigated by fluorescent protein labeling, and the effects of black phosphorus on cerebral ischemia/reperfusion injury were studied by 2,3,5-triphenyltetrazolium chloride (TTC) staining, neurobehavioral evaluation, and brain edema. Protein immunoblotting analysis was used to explore the molecular mechanism of the BP drug delivery system on ischemic brain injury. Hemolysis test and hematoxylin-eosin (H&E) staining were used to evaluate its biocompatibility. The results showed that BP had excellent drug loading capacity, uniform drug loading system structure and particle size, stable drug release curve, and excellent photothermal effect. Through the analysis and comparison of fluorescence intensity, it was found that BP can increase the permeability of blood-brain barrier (BBB) under the condition of near-infrared light assisted irradiation, and make drugs more pass through the BBB. In addition, the black phosphorus nano tablet drug delivery system can significantly improve the neurobehavioral disorder of mice after modeling, and the cerebral infarction area and brain edema degree are significantly decreased. Western blot experiments showed that the drug delivery system could play an anti-ischemic brain injury role by activating the expression of antioxidant signaling pathway proteins nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1). The hemolysis test and H&E test results of the BP drug carrier system showed that it had no obvious toxicity and high safety. In conclusion, the BP prepared in this study had high drug loading, good photothermal performance, and high safety. Under the near-infrared condition, they also have certain brain targeting ability, which can improve the therapeutic effect of drugs in the brain. Animal welfare and experimental procedures were following the regulations of the Animal Ethics Committee of the First Affiliated Hospital of Shihezi University.

Chinese Medicine in Treating Diabetic Renal Fibrosis： A Review / 中国实验方剂学杂志

Diabetic nephropathy （DN） is among the common microvascular complications of diabetes. In recent years， the incidence has been on the rise with the increase in prevalence of diabetes， threatening the health of human. The early stage of DN is characterized by excessive accumulation of extracellular matrix （ECM） and thickening of glomerular basement membrane which result in glomerular mesangial proliferation and massive collagen deposition. The late stage features glomerular sclerosis and renal fibrosis （RF）. It has been confirmed that RF is the key pathological process for the development of DN. Therefore， it is the research focus to explore the pathogenesis and treatment methods of RF. It has been frequently verified that Chinese medicine is superior in the treatment of diabetic RF. It relieves diabetic RF by regulating transforming growth factor-β （TGF-β）， secretory glycoprotein （Wnt）/β-catenin， nuclear factor-κB （NF-κB）， Notch， mitogen-activated protein kinase （MAPK）， and other signaling pathways. Therefore， this paper reviews the pathogenesis of diabetic RF and the treatment with Chinese medicine， which is expected to serve as a reference for clinical application of Chinese medicine in the treatment of diabetic RF.

Huidouba Ameliorates Kidney Oxidative Stress Injury by Down-regulating Nox4 Expression in Rats with Diabetic Nephropathy / 中国实验方剂学杂志

Objective:From a new perspective，to explore therapeutic effect of Huidouba (HDB) on alleviating kidney oxidative damage in rats with diabetic nephropathy (DN) and provide a scientific basis for developing HDB as a potential Tibetan medicine for treatment of DN. Method:Rats were fed with high-fat diet (HFD) and injected with streptozocin (STZ, 65 mg·kg-1) intraperitoneally to induce DN model, while rats in Blank group were injected with an equal volume of vehicle and fed with normal chow. The successfully modeling DN rats were randomly divided into three groups, 8 rats per group, DN model group (10 mL·kg-1·d-1), Metformin group (0.045 g·kg-1·d-1) and HDB group (0.18 g·kg-1·d-1). Monitor body weight (BW) and fasting blood glucose (FBG) weekly, and collect 24 hours urine before and after medication to examine microalbuminuria (mAlb). Calculate kidney index (KI) after sacrificing, analyze mAlb, serum creatinine (SCr) and blood urea nitrogen (BUN) with a fully automatic biochemical analyzer. Histopathology of kidney was observed by Masson staining. Lipid peroxidation malondialdehyde (MDA) assay kit was used to examine MDA content in kidney tissue. Nox4, as a subtype of triphosphopyridine nucleotide (NADPH) oxidase family was determined by Western blot and immunofluorescence assay of kidney tissue. Result:Compared with blank group, levels of FBG, 24 h mAlb, SCr, BUN and MDA in DN model group were increased (P<0.01), tissue damage was obvious and Nox4 expression in glumeruli was increased significantly (P<0.01). Compared with DN model group, levels of FBG, 24 h mAlb, SCr, BUN and MDA in drug administration groups were decreased (P<0.01), kidney injury was alleviated and Nox4 expression was down-regulated（P<0.01）. Conclusion:HDB as a Yiqiyangyin Tibetan medicine, could ease oxidative stress injury of kidney and reduce proteinuria in DN rats, thus prevent the development of DN. Its mechanism is closely related to down-regulating Nox4 expression of kidney tissue in DN rats.

SRGN′s interaction with TGFβ promote the chemoresistence of NSCLC / 中国医师杂志

Objective:To explore the role of small molecule glycoprotein Serglycin (SRGN) in chemotherapy resistance of non-small cell lung cancer (NSCLC).Methods:In NSCLC H1299 cell line, shRNA technology was used to interfere with the expression of SRGN and establish stable interfering cell line. Western blot and real time fluorescence quantitative polymerase chain reaction (qRT-PCR) were used to verify the knockdown efficiency; MTS was used to detect the knockdown cell line′s drug sensitivity to cDDP and Oxaliplatin; enzyme linked immunosorbent assay (ELISA), Western blot and qRT-PCR were used to explore the effect of transforming growth factor β (TGFβ) on SRGN and vice versa; Western blot was used to detect the effect of SRGN on epithelial-mesenchymal transition (EMT) related molecules, and online data bioinformatics was used to analyze the correlation between SRGN and EMT related molecules expression; in addition, online prognostic analysis software (kmplot) was used to analyze the correlation between SRGN, TGFβ and prognosis of lung cancer patients.Results:Comparing with the control group, the test group, knocking down SRGN can obviously improve the drug sensitivity of NSCLC cell to cDDP ( P=0.032 7) or Oxaliplatin ( P=0.014 2). TGFβ can enhance the experission of SRGN in NSCLC and SRGN also can help TGFβ secreted from cells. SRGN promotes the epithelial mesenchyme transition by modulating Snail1. By analyzing TCGA database, we found that the expression of SRGN was negatively correlated with the expression of CDH1 (coding for Ecadherin protein) ( r=-0.25) and there was a positive correlation with Snai1 expression ( r=0.37). These results suggest that SRGN can promote the change of EMT in lung cancer cells through TGF β 1 and snail 1. The overall survival time of NSCLC patients with low expression of SRGN was much longer than the patients with high expression of SRGN ( P=0.007 7). The overall survival time of NSCLC patient with low expression in both SRGN and TGFβ1 or TGFβ2 was 73months or 42.8 months longer than that with high expression in both SRGN and TGFβ1/2. Conclusions:Intercting with TGFβ1, SRGN promotes EMT of NSCLC cells, which facilitates the chemoresistence of NSCLC. The simultaneous low expression of SRGN and TGFβ1 or TGFβ2 can significantly prolong the overall survival of patients with NSCLC.

Tumor-associated macrophage enhances invasion and migration of breast cancer cells T47D / 中国医师杂志

Objective To explore the effect of tumor-associated macrophage (TAM) on invasion and migration of breast cancer cells T47D.Methods Briefly,phorbol 12-myristate 13-acetate (PMA) was employed to treat mononuclear cells THP-1.And then interleukin 4 (IL-4) was applied to stimulate mentioned cells to establish activated TAM.Furthermore,the supernatant of M0 and M2 was used to culture T47D cells,respectively.Then,the wound healing experiment and transwell assay were carried out to investigate cells invasion and migration.Finally,epithelial-mesenchymal transition (EMT) biomarkers in T47D cells treated with M0 or M2 were determined by quantitative real time polymerase chain reaction (qRT-PCR) and western blot assays.Results Activated TAM model was successfully established by PMA following with IL-4.Comparing with M0,M2 evidently accelerated invasion and migration in T47D cells.Meanwhile,M2 upregulated N-cadherin,vimentin and β-catenin,downregulated E-cadherin at mRNA and protein expression levels.Conclusions M2 promotes invasion and migration of T47D cells via EMT.

Effects and mechanisms of histone deacetylase inhibitors on invasion and metastasis of Her2 positive breast cancer cell / 中国医师杂志

Objective To investigate the effect of histone deacetylase inhibitor on Her2 positive breast cancer cell line BT474 and SKBR3 in apoptosis and metastasis.Methods Histone deacetylase inhibitor MS-275,suberoylanilide hydroxamic acid (SAHA)(4 μmol/L,and 50 μmol/iL,respectively) treated the cell lines BT474 and SKBR3 cells.Flow cytometer examined the apoptosis ratio.Transwell tested their metastatic activity.Western blot assay was performed to detect the associated proteins.Results SAHA and MS-275 inhibited the cell survival.The BT474 cell survival was (39 ± 11) ％,(54 ± 8) ％,and the SKBR3 survival was (62 ± 6) ％,(71 ± 9) ％,according to the fluorescence-activated cell sorting (FACS) result.SAHA and MS-275 induced the BT474 cell apoptosis 8.46± 0.28 (P ＜0.01),4.15 ± 0.71 (P ＜0.01) fold change,respectively;and upregulated the SKBR3 cell apoptosis ratio 5.51 ± 1.24 (P ＜0.01),4.04 ±0.69 (P ＜0.01) fold.The Transwell result showed that SAHA,MS-275 inhibited the Transwell ability of BT474 from the control 184.7 ± 18.8 to 104.3 ± 7.1,131.3 ±9.1 per view,and the SKBR3 from control 60 ± 16.7 per view to 14.3 ± 6.5,34.3 ± 8.7 per view.The Western blot result showed that SAHA,MS-275 inhibited the protein level of vimentin,Her2,β-catenin,histone deacetylase inhibitor (HDACi),and upregulated the acetylation level of histone 3.The E-caherin protein was regulated in BT474 and SKBR3 cells.Conclusions MS-275,SAHA can induce BT474 and SKBR3 apoptosis significantly,also inhibit their metastatic activity.

Lower respiratory tract infection caused by influenza virus A and influenza virus B in Wenzhou, China: a clinical analysis of 366 children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To compare the epidemiological and clinical features of lower respiratory tract infection (LRTI) caused by influenza virus A (IVA) and influenza virus B (IVB) in children.</p><p><b>METHODS</b>The clinical data of 366 children with LRTI caused by influenza virus (IV), who were hospitalized in Yuying Children′s Hospital of Wenzhou Medical University between 2010 and 2014, were analyzed retrospectively, and there were 272 cases caused by IVA and 94 cases caused by IVB.</p><p><b>RESULTS</b>IV was mainly prevalent from December to March of the next year, with the predominance of IVA. There were small peaks of IVA prevalence in July or September every other year, and IVB was prevalent from December to March of the next year every other year. The children with LRTI caused by IVA alone had a significantly higher white blood cell (WBC) count and significantly higher percentages of children with increased WBC, abnormal serum sodium, and abnormal serum potassium than those caused by IVB alone (P<0.05). However, there were no significant differences in age, sex, underlying diseases, clinical manifestations, and co-infection rate with bacteria or atypical pathogens between the two groups (P>0.05). The rate of co-infection with respiratory syncytial virus (RSV) was significantly higher in the IVB group than in the IVA group (P<0.01).</p><p><b>CONCLUSIONS</b>IVA is prevalent in winter and spring every year and has small peaks in summer every other year, while IVB is prevalent in winter and spring every other year. Compared with IVB, IVA causes more cases of increased WBC and electrolyte disturbance. The children infected with IVB are more likely to be co-infected with RSV. The children with LRTI caused by IVA and IVB have similar clinical manifestations.</p>

The effect of HDAC inhibitor SNDX-275 on inhibiting breast cancer BT474 cell proliferation / 国际肿瘤学杂志

Objective To explore the effect and molecular mechanism of HDAC inhibitor SNDX-275 inhibiting cell proliferation in ErbB2-overexpressing breast cancer BT474 cells.Methods Breast cancer BT474 cells were treated with HDAC inhibitor SNDX-275,setting as test group,and the cell line treated with phosphate buffered saline (PBS)as control.The concentration of SNDX-275 were 0,0.5,1 .0,2.0,3.0,4.0 μmol/L respectively.Cell proliferation was analyzed by MTS assay and colony formation assay,the expressions of ErbB2, ErbB3,p-Akt were analyzed by Western blotting,and the expressions of miR-125a,miR-125b were analyzed by RT-PCR.After transfecting miRNA125 inhibitor into BT474 cells,the inhibition rate of SNDX-275 was tested by MTS assay .Results MTS result showed that SNDX-275 inhibited cell proliferation in BT474 cells in a dose-dependent manner.The inhibition rate of 4.0 μmol/L SNDX-275 was about (68.00 ±4.45)%.Clone assay indicated SNDX-275 could inhibit the proliferation of BT474 cells.Western blotting result indicated that SNDX-275 significantly inhibited the protein expressions of ErbB2,ErbB3 and p-Akt,RT-PCR result illustrated 2 μmol/L SNDX-275 could increase the expressions of miR-125a and miR-125b about 3.22 ±1 .17,5.42 ±0.38 times compared with the PBS control respectively,the difference has a statistical significance (t =4.338,P =0.049;t =21 .805,P =0.002).MTS result indicated that compared with the PBS control,the inhibition rate of SNDX-275 group was (56.97 ±3.56)%,while the inhibition rate of SNDX-275 and miRNA125 inhibitor group was (10.67 ±2.21 )%,with a statistical significance(t =-10.993,P =0.008).Conclusion SNDX-275 could inhibit cell proliferation of ErbB2-overexpressing breast cancer BT474 cells,by inhibiting ErbB2-ErbB3-Akt sig-nal pathway through up-regulating miR-125a and miR-125b.

Expression of caspase-3 and HAX-1 after cerebral contusion in rat / 法医学杂志

OBJECTIVE@#To observe the expression pattern of caspase-3 and HCLS1-associated protein X-1 (HAX-1) at different time after cerebral contusion in rat, and explore the new method for estimating the injury interval.@*METHODS@#The cerebral contusion model was established using adult SD male rats. Then the rats were randomly allocated into 8 groups: 2 h, 6 h, 12 h, 1 d, 3 d, and 7 d after cerebral contusion, sham-operation and normal control. Expression of caspase-3 and HAX-1 protein after cerebral contusion in rat was detected by Western blotting. Laser scanning confocal microscope was used to observe the number of HAX-1 positive cells and TUNEL-stained cells after cerebral contusion.@*RESULTS@#The expression of caspase-3 increased parallelly with the time after cerebral contusion and reached the peak value on 3 d. The expression of caspase-3 decreased gradually and still maintained a high level expression on 7 d (P < 0.05). The expression of HAX-1 positive cell went up after injury, and reached the peak value at 6 h (P < 0.05), then turned down gradually after 12 h and went out of detection after 3 d. The number of TUNEL-stained cells increased obviously at 2 h and reached the peak value on 3 d. The number of TUNEL-stained apoptotic cells decreased gradually and still maintained a high level expression on 7 d (P < 0.05).@*CONCLUSION@#The expression of caspase-3 and HAX-1 after cerebral contusion has time sequential regularity, which may provide new evidence for forensic diagnosis of cerebral contusion interval.

miR-34c suppresses cell proliferation and invasion of prostate cancer PC3 cells / 中国医师杂志

Objective To explore the effect and molecular mechanism of miR-34c in cell proliferation and invasion of human prostate cancer PC3 cells.Methods miR-34c was synthesized and transfected into prostate cancer PC3 cells.The ability of cell pro liferation was examined with methyl thiazolyl tetrazolium (MTT),and invasion with Transwell assay.The level of c-met mRNA was analyzed by real-time reversing transcription polymerase chain reaction (RT-PCR).Western blot was used to detect the expressions of cmet,cyclin D1,and EMT-associated markers.Results Over-expression of miR-34c reduced PC3 cell proliferation,and inhibited the abilities of invasion.Bioinformatics analysis showed that 3'-UTR of c-met had two miR-34c matched sites,and miR-34c inhibited the expressions of c-met mRNA and protein.Meantime,miR-34c obviously decreased the expressions of cyclin D1 and N-cadherin,but increased the expression of E-cadherin in PC3 cells.Conclusions miR-34c may suppress cell proliferation and invasion in prostate cancer PC3 cells,by which mechanism is related to reduction of the expressions of c-met,cyclin D1,N-cadherin,and upregulation of E-cadherin.

Investigation on low doses of metformin induce hepatoma cells senescence and its underlying mechanism / 中国基层医药

Objective To explore the effect of metformin on hepatoma cells senescence and the underlying mechanism.Methods Cell proliferation,cycle and apoptosis were examined by MTS and flow cytometry assay in response to different concentrations of metformin(0,0.01,0.1,1,10 and 50mmol/L).Senescence-associated β-galactosidase (SA-β-ga1) staining and senescence marker Dec1 protein levels were used to evaluate the effect of metformin on hepatoma cells senescence.In addition,protein expression of p-AMPK,p-ACC and AMPK was detected by Western blot analysis.Results Metformin suppressed proliferation of HepG2 cells in a dose-dependent manner.High concentrations of metformin (10 and 50mmol/L) promoted cell apoptosis,while lower doses of metformin (0.01,0.1 and 1mmol/L) led to enlarged and flatten senescent morphology and increased SA-β-ga1 positive cells.Moreover,cell cycle was blocked in G0/G1 phase and protein levels of senescent marker Dec1,p-AMPK and p-ACC were significantly enhanced,whereas AMPK protein expression was almost unchanged.Conclusion We showed here that high dose of metformin promotes HepG2 cells apoptosis,but low doses of metformin induce cellular senescence,which may be related to the activation of AMPK signaling.These data will provide vital evidence for improving the outcome of comprehensive treatment in HCC patients by driving hepatoma cells to undergo senescence.

MicroRNA181b/c inhibits the metastasis of breast cancer by targeting glycoprotein serglycin / 中国医师杂志

Objective To investigate the relationship of glycoprotein serglycin (SRGN) expression with invasion and metastasis of breast cancer cells,and the role of microRNA in the regulation of SRGN expression.Methods Real-time quantitative polymer ase chain reaction (PCR) and Western blot were used to detect the differences in SRGN expression between higher metastasis Michigan cancer foundation-7 (MCF-7)/5-Fu breast cancer cell lines and weaker metastasis MCF-7 cell line.The siRNA interference experiment and in vitro Transwell experiment were used to detect effect of SRGN on the ability of invasion and metastasis of breast cancer cells.Bioinformatics software was used to predict miRNAs targeting SRGN,and integrated microRNA differentially expressed chip data between breast cancer cell MCF-7 versus MCF-7/5-Fu.The miRNA quantitative PCR was used to determine the differences of candi date miRNA expression.After transfection of microRNA minics,Western blot was used to test candidate microRNA target SRGN.Transwell experiment was used to test the effects of candidate microRNAs on tumor cell invasion and metastasis.Results SRGN was increased significantly in MCF-7/5-Fu cells,and the invasion and metastasis of tumor cells were inhibited when SRGN was interfered.In addition,miR181 b/c expressed in MCF-7/5-Fu cells was reduced significantly,negatively correlated with SRGN expression,and targeted SRGN expression.It inhibited invasion and metastasis of tumor cells.Conclusions MicroRNA181b/c inhibits metastasis of breast cancer by targeting SRGN.

Prospective study of the association between fasting glucose concentrations and liver cirrhosis risk / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the association between concentration levels of fasting serum glucose and liver cirrhosis.</p><p><b>METHODS</b>A nested case-control study was carried out based on the sample cohort from the Nutrition Intervention Trials previously conducted in one country in Henan province. Using an automatic biochemical analysis system and enzyme-linked immunoassay, baseline serum samples from 310 liver cirrhosis patients and 620 healthy controls were tested for fasting glucose concentration, hepatitis B surface antigen (HBsAg), hepatitis B core antibody (anti-HBc), and hepatitis C virus antibody (anti-HCV). Baseline demographic information was collected by questionnaire. The serum glucose values were divided into quintiles and applied to a logistic regression model to estimate odds ratios (ORs) and 95% confidence intervals (95% CIs).</p><p><b>RESULTS</b>The mean fasting blood glucose level was significantly higher in cases (4.5+/-1.8 mmol/L) than in controls (4.2+/-2.1 mmol/L) (t=-2.414, P=0.016). The individuals in the highest quintile had a significantly higher risk of disease than those in the lowest quintile [OR=1.672 (1.080, 2.588)]. Moreover, increase in glucose level was accompanied by increased risk, and the relation showed statistically significant linearity (P=0.002). The statistical significance of risk remained after adjustment for potential confounders, including sex, age, HBsAg, anti-HBc, and residence running water status [OR=1.96 (1.216, 3.157), P=0.001].</p><p><b>CONCLUSION</b>Elevated serum fasting glucose concentration was an independent risk factor of cirrhosis.</p>

Time-order expression of caspase-3 and iNOS in contused human brain tissue / 法医学杂志

OBJECTIVE@#To investigate the expression of caspase-3 and iNOS in different intervals and to provide evidence for estimation of injury intervals after brain contusion in human.@*METHODS@#Thirty cases died of serious brain injury were included into the injury groups and 5 cases died of non-brain injury were served as control group. To analyze the changes of caspase-3 and iNOS expression in brain samples at different intervals (2h, 4-8h, 10-14h, 1-2d, 3-5d, 8-11d) by immunohistochemistry and auto-image analysis system.@*RESULTS@#The level of caspase-3 expression started to increase in 2 hours after brain contusion compared to the control group (P<0.05). The level of caspase-3 expression continued to increase in 1-2 days and maintained high level in 3-5 days compared to the control group (P<0.05), then decreased gradually. There was no statistically significant difference between the expression level of iNOS in 2 hours with the control group (P>0.05). But the expression level of iNOS began to increase in 4-8 hours after brain contusion and reached its maximum in 1-2 days, then decreased. Weak expression of iNOS still could be detected in 8-11 days.@*CONCLUSION@#The expression of caspase-3 and iNOS can be used as effective evidence for human brain contusion interval.