Objective To investigate the effect of different ratios of ginsenosides(GS) and panax notoginsenosides(PNS),which are extracted from Qi-strengthening herbs of Radix Ginseng and blood-activating herbs of Radix Notoginseng respectively,on rats peritoneal mesothelial cells(PMCs) in peritoneal dialysis solution(PDS).Methods PMCs were isolated from rat peritoneal membrane by trypsin digestion method,and then a stable PMCs culture model was established.PMCs were pre-exposed in 4.25% PDS for 3 hours,and then respectively grew in culture solution with different ratios of GS or PNS for 6 hours.The capacity of proliferation of rat PMCs was assessed by methyl thiazolyl tetrazolium(MTT) assay.Results GS 100?g/mL and PNS 80?g/mL showed protective effect on the proliferation of injured PMCs in PDS(P 0.05).Conclusion The combination of GS and PNS,which are extracted from Qi-strengthening herbs of Radix Ginseng and blood-activating herbs of Radix Notoginseng respectively,exerts certain protective effect on PMCs in peritoneal dialysis solution.

Objective To evaluate the value of detecting chlorophyll related genes of plankton in the diagnosis of drowning. MethodsEighteen rabbits were divided randomly into three groups: death by drowning (n=10), postmortem submersion (n=6) and control (n=2). The heart blood, lung, liver, kidney and brain tissues were taken from every rabbit. After isolated plankton from tissues with percoll and extracted their DNA, the chlorophyll-related genes, including EG (EG1 and EG2) and SK (SK1 and SK2), were detected using PCR technique. Meanwhile, diatom test was also performed from lung and liver tissues by nitric acid digestion method. ResultsFor the drowning group, the specific amplification products for EG1 were detected from 9 samples in heart blood, 10 samples in lung, 9 samples in liver, 7 samples in kidney and 8 samples in brain. The products for EG2 were detected from 8 samples, 10 samples, 7 samples, 5 samples and 7 samples accordingly. There were a small number of positives in heart blood, lung and kidney with SK1 and SK2 (≤2). For the postmortem submersion group, only one case was positive from heart blood and lung tissue respectively for EG1. No amplified product was detected for EG1 and EG2 in various tissues in control group, and also no product was detected for SK1and SK2 in other groups. In addition, diatoms were detected from 9 lung and 3 liver tissues in drowning group with the nitric acid digestion, and only one sample of lung was positive in the postmortem submersion group. ConclusionThe detection rate of the chlorophyll-related gene EG with PCR method was higher than that of diatom with nitric acid digestion method in drowning victims, and it can be used as a potentially useful tool for diagnosing drowning.

Objective To develop a simplified bisulfite genomie sequencing(BGS)method for DNA methylation marker scanning.Methods According to modified BGS protocol,the desalt DNA treated with bisulfite were directly used for bisulfite-PCR(BSP)without alkali treatmenL Complement of the bisulfite modification Wag accomplished by a prolonged pre-denaturation stage.After BSP,a second round PCR was performed with a pair of GC tagged primers to adjust the GC content of the amplieon for direct sequencing.To assess this improved protocol,promotor methylation of TNF-α gene in 3T3-L1 cell and androgen receptor(AR)gene in Hela cell was investigated.The real time BSP for Alu was also used to compare the sensitivity of the modified assay with traditional assay.Results Both the hypermethylated TNF-α promotor and hypomethylated AR promotor were successfully sequenced by improved BGS method,and the results were consistent with that of the traditional assay.The conversion rate reached 100%,while the conversion specificity was higher than 93.75%.The sensitivity of improved BGS method inereaged significantly(t=2.978 2,P＜0.05)and showed good reproducibility.Condusion The improved BGS method is simple and sensitive,facilitating more ambitious genomic methyhtion mapping studies.

Objective To studying the MR findings and pathology of peripheral small intrahepatic cholangiocarcinoma and improving the understanding of peripheral small cholangiocarcinoma with no-bile duct dilatation. Methods A retrospective analysis of 12 patients with intrahepatic peripheral cholangiocarcinoma which were confirmed by surgery and pathology, all patients were examined by abdominal MRI without and with contrast. Correlation was made with gross pathology and surgical pathological specimen. Results On T1WI, there were 4 cases of complex low signal intensity and 8 cases of low signal intensity. On T2WI, there were 8 cases of high signal intensity and 4 cases of complex high signal intensity. Enhanced MRI showed: marked nidus enhancement on arterial phase in 1 case, and the pathological diagnosis was poorly differentiated adenocarcinoma. Inhomogeneous enhancement or annular enhancement were seen in 10 cases on arterial phase, 3 of these cases showed thin annular enhancement on arterial phase, low signalintensity on portal venous phase and isointensity on delayed phase. One case showed delayed enhancement. Thick circular enhancement correlated with pathological changes of survival of tumor cells, center areas correlated with fibrous connective tissue, and a small amount of necrotic tissue. Island-like enhancement or inhomogeneous enhancement were seen in 3 cases. Corresponding pathological changes consisted of tumor tissue and a small amount of fibrous connective tissue, as well as somenecrotic tissue. In 1 case, no enhancement was seen on all three phases and pathological changes showed cystic changes, hemorrhage, necrosis, with survival tumor cells seen between cyst and normal liver tissue. Conclusions MRI scanning of peripheral small cholangiocarcinoma lacked characteristic features, but dynamic contrast-enhanced MR had certain specific findings. Due to different pathology, the fibrous tissue, necrotic tissue and survival tumor tissue components were exhibited different imaging findings.

Objective To investigate the application value of dual-layer spectral computed tomography(DLCT)45 keV single energy image in follow up after transcatheter arterial chemoembolization(TACE)for hepatocellular carcinoma(HCC).Methods The DLCT images of 60 patients with HCC after TACE treatment were analyzed retrospectively.The CT value and standard deviation(SD)value of the lesion region of interest(ROI)and surrounding normal liver parenchyma on two kinds of images with 45 keV energy spectrum CT and 120 kVp conventional CT were measured respectively,then the contrast-to-noise ratio(CNR)between the lesions and surrounding normal liver parenchyma was calculated,and their differences were compared.The image quality of the two kinds of images was scored subjectively with the 3-point method,and the patients were divided into different groups according to the enhance-ment degree and tumor staining.The receiver operating characteristic(ROC)curve was also drawn.Results(1)There was no signif-icant difference in CNR between arterial phase,portal phase,and delayed phase of 120 kVp conventional CT(Hc=1.128,P>0.05).The CNR of 45 keV energy spectrum CT was higher than that of 120 kVp conventional CT,with a statistically significant difference(Z=5.060,P<0.05).(2)The subjective score of 45 keV energy spectrum CT was higher than that of 120 kVp conventional CT,and the difference was statistically significant(Z=5.335,P<0.05).ROC curve analysis showed that 45 keV energy spectrum CT had a larger area under the curve(AUC)than 120 kVp conventional CT,and the difference was statistically significant(Z=4.136,P<0.001).Conclusion 45 keV energy spectrum CT image can significantly improve the CNR between tumor and normal liver parenchyma,and it has the better image quality and higher diagnostic efficiency.

Background Mitochondrial dynamin-related protein 1 (DRP1) regulates mitochondrial division and plays an important role in maintaining hepatocyte function. However, the role of DRP1 in cadmium exposure-induced maternal liver damage in pregnant mice remains unclear. Objective To investigate the role and mechanism of DRP1 in maternal liver damage induced by cadmium exposure during pregnancy. Methods This study consisted of animal experiments and cell experiments. (1) Animal experiments. Mice at 14 days of gestation were randomly divided into three groups: a control group, a low-dose cadmium group (LCd group: 2.5 mg·kg⁻¹), and a high-dose cadmium group (HCd group: 5 mg·kg⁻¹). The pregnant mice were intraperitoneally injected with cadmium chloride (CdCl₂) for 6 and 24 h in the next morning. The weights of pregnant mice, uterus, maternal liver, and fetal mice were recorded after sacrifice. Serum and liver of pregnant mice were collected, the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were detected, and liver tissues were stained with HE to observe changes in liver function and liver tissue structure. The expressions of oxidative phosphorylation-related proteins, hypoxia inducible factor-1α (HIF-1α) and DRP1 proteins in liver of pregnant mice were detected by Western blotting. (2) Cell experiments. AML12 cells were treated with CdCl₂ (10 μmol·L⁻¹) for 0, 2, 6, 12, and 24 h. The expressions of oxidative phosphorylation-related proteins, DRP1, and hypoxia inducible factor-1α (HIF-1α) proteins were detected. AML12 cells were pretreated with DRP1 inhibitor Mdivi-1 for 1 h and then CdCl₂ (10 μmol·L⁻¹) for 12 h to detect the expression of oxidative phosphorylation-related proteins and DRP1 protein. AML12 cells were treated with Hif-1α siRNA for 48 h and CdCl₂ (10 μmol·L⁻¹) for 6 h to detect the expression of HIF-1α and DRP1 proteins. Results The results of animal experiments showed that cadmium exposure in pregnant mice had no effects on maternal liver weight and liver coefficient. However, the histomorphological changes and necrosis in hepatocytes were observed. Compared with the control group, the serum ALT and AST levels of pregnant mice in the LCd group were significantly increased after 6 h (P＜0.05), and the levels in the HCd group were significantly increased after 6 and 24 h (P＜0.05). Cadmium exposure during pregnancy significantly up-regulated HIF-1α and DRP1 expressions and down-regulated the expressions of oxidative phosphorylation-related proteins in maternal livers. In vitro cell experiments showed that the expressions of oxidative phosphorylation-related proteins was significantly decreased and HIF-1α and DRP1 protein expressions were significantly increased in the AML12 cells treated with CdCl₂ for 6 h. Mdivi-1 pretreatment significantly antagonized the inhibitory effect of cadmium on the expressions of oxidative phosphorylation-related proteins in AML12 cells, while Hif-1α siRNA pretreatment significantly antagonized the up-regulative effect of cadmium on DRP1 expression in AML12 cells. Conclusion Cadmium exposure in pregnant mice may up-regulate DRP1 expression by activating HIF-1α signaling, then inhibit oxidative phosphorylation level of hepatic cells, and ultimately lead to maternal liver damage.