Effects of electroacupuncture on CYP7A1 expression in liver of rabbits with atherosclerosis / 中国针灸

OBJECTIVE@#To observe the effects of electroacupuncture (EA) at "Neiguan" (PC 6), "Guanyuan" (CV 4) and "Zusanli" (ST 36) on CYP7A1 expression in liver of rabbits with atherosclerosis (AS), and to explore the mechanism of acupuncture for prevention and treatment of atherosclerosis.@*METHODS@#A total of 26 male rabbits were adaptively fed for 1 week in different cages. Seven rabbits were randomly divided into a blank group, and the remaining 19 rabbits were divided into a model group. The blank group was fed with normal diet, while the model group was fed with high-fat diet. After high-fat diet for 4 weeks, the rabbits in the model group were treated with balloon injury surgery of the common carotid artery; after surgery, the rabbits were fed with high-fat diet for 4 weeks. One rabbit was randomly selected from the blank group and model group to obtain the pathological section of carotid artery; the HE staining was used to observe the pathomorphology of atheromatous plaque to determine the success of modeling or not. After successful establishment of modeling, 18 rabbits were randomly divided into a AS model group, an EA group and a medication group, 6 rabbits in each one. The rabbits in the AS model group received no treatment; the rabbits in the medication group were treated with intragastric administration of atorvastatin calcium tablets; the rabbits in the EA group were treated with EA at "Neiguan" (PC 6), "Guanyuan" (CV 4) and "Zusanli" (ST 36), once a day, 20 min per treatment; six-day EA treatment constituted one course, and totally 4 courses were given with an interval of 1 day between courses. After treatment, vein blood was collected from rabbit ear, and cholesterol (CHO), triglyceride (TG), low-density lipoprotein (LDL) and high-density lipoprotein (HDL) were measured in each group; the CYP7A1 protein expression in rabbit liver was measured by Western blot method, and CYP7A1 mRNA expression in rabbit liver was measured by RT-PCR.@*RESULTS@#Compared with the blank group, the contents of CHO, TG and LDL in the AS model group were significantly increased, but HDL was significantly decreased, and the expression of CYP7A1 protein and CYP7A1 mRNA in the liver were significantly decreased (all 0.05).@*CONCLUSION@#EA could significantly improve blood lipid and promote the expression of CYP7A1 mRNA in rabbits with atherosclerotic, which may be one of the mechanisms of EA for atherosclerosis.

The protective effect of Qizhi hypoglycemic tablet on foot ulcer in streptozotocin-induced diabetes in rats / 药学学报

This study aimed to determine the protective effect of Qizhi hypoglycemic tablet (QZHGT) on foot ulcer in diabetic rats and explore its possible mechanism. Diabetes was induced by streptozotocin injection in rats. The rats received QZHGT (780 mg·kg-1), metformin hydrochloride tablet (Metf, 200 mg·kg-1) or glibenclamide tablet (Glib, 1.5 mg·kg-1) alone via intragastric administration once a day for three months. Food ulcer was prepared by foot skin excision after drug therapy lasted for two months, and the dynamic changes in food ulcer healingwere determined. During the experiment, blood glucose, serum levels of vascular endothelial growth factor (VEGF) and inducible nitric oxide synthase (iNOS), factor Ⅲ (FⅢ) and four coagulation parameters [thrombin time (TT), activated partial thromboplatin time (APTT), prothrombin time (PT), fibrinogen (FIB)] were detected. Finally, the protective mechanisms of QZHGT against diabetes and foot ulcer were analyzed by network pharmacology, and immunohistochemistry was used to confirm the expression of transforming growth factor-β (TGF-β) and nuclear factor κB (NF-κB) in pancreatic tissue. All animal procedures were approved by the Animal Experimentation Ethics Committee of Henan University (permission number HUSAM 2016-288). The results showed that the lasting hyperglycemia, polydipsia, polyphagia, polyuria and body weight lost took place in model rats compared to those in normal rats. These model rats also showed an increase in serum VEGF and iNOS, FⅢ, TT, APTT and PT, and a reduction in FIB and wound healing. Metf or Glib significantly improved hyperglycemia, polydipsia, polyphagia, polyuria and emaciation, but failed to ameliorate hypercoagulation and wound healing. QZHGT showed a similar effect on polydipsia, polyphagia, polyuria and emaciation to Metf or Glib, although it was inferior to them in hypoglycemic action. Importantly, QZHGT significantly improved hypercoagulation and wound healing, and attenuated serum VEGF and iNOS. Network pharmacology revealed that QZHGT decreased hyperglycemia through "insulin resistance pathway", improved coagulation status through "HIF-1 signaling pathway", prevented diabetic foot ulcers through "VEGF signaling pathway", "MAPK signaling pathway" and "NF-κB signaling pathway". Immunohistochemistry showed that QZHGT could inhibit the expression of TGF-β and NF-κB in pancreatic tissue to maintain islet function in diabetic rats. In summary, these data suggest that QZHGT can prevent pancreatic injury for adjunctive hypoglycemia and diabetic foot ulcer treatment, and is a better preventive and therapeutic drug for diabetic foot ulcer.

Study on Helicobacter infection in liver tissue from hepatocellular carcinoma / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate the relationship between Helicobacter species and hepatocellular carcinoma(HCC).</p><p><b>METHODS</b>Liver samples resected during operation from 34 patients with HCC diagnosed by histopathology and 20 without primary liver carcinoma as controls were studied. The two groups of sample were cranked out pathologic slice for in situ hybridization of Helicobacter, Helicobacter pylori and Helicobacter hepaticus. Qualitative and quantitative studies were used to assess the correlation of liver tissue Helicobacter infection with HCC.</p><p><b>RESULTS</b>64.71% (22/34). of the samples of HCC showed positive for Helicobacter specific 16S rRNA-mRNA gene by in situ hybridization, while none was positive in controls (P < 0.01).</p><p><b>CONCLUSION</b>Helicobacter pylori were found in the liver of patients with HCC.</p>

Synergism between a siRNA targeted to survivin and 5-FU in inhibiting MCF-7 cell proliferation in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the role of small interfering RNA (siRNA) targeted to survivin in combination with 5-fluorouracil (5-FU) in inhibiting the proliferation of MCF-7 cells.</p><p><b>METHODS</b>A siRNA targeted to survivin was synthesized and transfected into MCF-7 cells via lipofectin. Changes of the cell growth activity in response to combined treatment with survivin siRNA and 5-FU or 5-FU treatment alone was evaluated by MTT assay. The Q method of Jin Zhenjun was used to evaluated synergism between the synthesized siRNA and 5-FU.</p><p><b>RESULTS</b>Treatment with 5 nmol/L siRNA reduced the IC50 of 5-FU from 4.42 to 1.18 microg/ml, and the inhibitory effect of combined treatment on MCF-7 cells was higher than that of 5-FU alone (F=26.74, P<0.01). Synergism effect was observed between 5-FU at lower concentrations and survivin siRNA.</p><p><b>CONCLUSION</b>siRNA may enhance the effectiveness of 5-FU in inhibiting the proliferation of MCF-7 cells.</p>

Study on loss of heterozygosity and microsatellite instability in hepatocellular carcinoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the role of loss of heterozygosity (LOH) in tumor suppressor genes (TSG) and microsatellite instability (MSI) in hepatocarcinogenesis, as well as their correlation with clinicopathologic features.</p><p><b>METHODS</b>LOH in 6 TSG (APC, DCC, MCC, OGG1, p53 and RB1) was detected in 36 informative cases of hepatocellular carcinoma (HCC), among 92 surgically resected HCC. Thirteen polymorphic microsatellite markers were also studied in 15 of these cases by microdissection-based PCR amplification and direct DNA sequencing. The correlation between genetic alterations and clinicopathologic features was analyzed.</p><p><b>RESULTS</b>The overall incidence of LOH in HCC was 41.7% (15/36). There was no LOH in MCC gene. 46.2% (6/13) microsatellites showed LOH in 9 of the 15 cases of HCC (60%). Certain clinicopathologic differences were observed between cases (number = 7) with LOH in APC, OGG1 and DCC ("type I") and cases (number = 8) with LOH in p53 and RB1 ("type II"). The mean tumor size of these two types was 2.9 (+/- 1.7) cm and 7.2 (+/- 3.4) cm, respectively (P < 0.01); and the mean survival was 72.0 (+/- 38.6) months, and 51.0 (+/- 30.4) months, respectively (P < 0.05).</p><p><b>CONCLUSIONS</b>Compared with MSI pathway, LOH pathway plays a more important role in the development of HCC. A multistep hepatocarcinogenesis is likely, with LOH in APC, OGG1 and DCC ("type I") being an early event and LOH in p53 and RB1 ("type II") being a late event. On the other hand, MCC gene seems to play no role in the whole process.</p>

Pathologic diagnosis of 1123 post-transplant liver biopsies from 665 liver transplant patients / 中华病理学杂志

<p><b>OBJECTIVE</b>To summarize the Chinese experience in pathologic diagnosis of liver biopsies after orthotopic liver transplantation (OLTx).</p><p><b>METHODS</b>1123 post-transplant liver biopsies from 665 OLTx patients from the Shanghai Eastern Hepatobiliary Surgery Hospital, Tianjin First Central Hospital, Guangzhou Sun Yat-sen University and Chongqing Southwest Hospital were retrospectively analyzed. All liver biopsies were stained with hematoxylin and eosin. Immunohistochemical studies for cytomegalovirus, HBsAg, CK19, CD4 and CD8 were also performed in selected examples.</p><p><b>RESULTS</b>In the involved hospitals, 4 to 12 types of complications were encountered after OLTx. The number of liver biopsies performed for each patient ranged from 1 to 9 (mean = 2.2). The timing of these biopsies varied from the second to the 2877 th post-transplant day. The 5 most common complications were acute cellular rejection (35.6%), ischemic-reperfusion injury (13.4%), biliary stricture (5.6%), drug complication (5.0%) and chronic rejection (4.7%). The 5 earliest complications after OLTx were primary non-function (occurring at day 4.7 +/- 2.1), ischemic-reperfusion injury (occurring at day 14.0 +/- 4.0), acute cellular rejection (occurring at day 32.1 +/- 62.9), hepatic artery thrombosis / stricture (occurring at day 62.9 +/- 74.2) and cytomegalovirus infection (occurring at day 107.7 +/- 93.0).</p><p><b>CONCLUSIONS</b>This study has evaluated the types, incidence and timing of major complications occurring after OLTx. The most important issue is the distinction between rejection and non-rejection pathology. Thorough understanding of atypical pathologic features of these complications is necessary. The Banff Schema (rejection activity index) for grading liver allograft rejection is useful for monitoring anti-rejection therapy and should be used routinely.</p>

Features of micro satellite alterations on chromosome 4 in hepatocellular carcinoma / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the features of micro satellite alterations and their association with clinicopathological characteristics of hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>Ten high-polymorphic micro satellite markers on chromosome 4 were selected to be detected for loss of heterozygosity (LOH), micro satellite instability (MSI) and allelic imbalance (AI) in 56 HCC using PCR-simple sequence length polymorphism (PCR-SSLP) analysis.</p><p><b>RESULTS</b>LOH was found in 40 of 56 HCC (71.4%) on at least 1 locus, the top two loci were D4S426 (61%), D4S1534 (53.7%). LOH on D4S406 was significantly higher in cases with positive serum HBsAg than in those with negative HBsAg. Similarly, LOH on D4S1538 occurred more frequently in patients with HBsAg negative than those with HBsAg positive [76.9% (20/26) vs 12.5% (2/16), chi2=13.999, P<0.01]. LOH on D4S426, D4S1615 and D4S165 were more frequent in poorly or moderately differentiated HCC than in well-differentiated HCC [76.7%(23/30) vs 18.2%(2/11), chi2=9.242, P<0.01; 53.8% (14/26) vs 16.7% (2/12), P<0.05; 60.7% (17/28) vs 18.2% (2/11), P<0.01]. LOH on loci D4S2921 was more frequently detected in tumors with intrahepatic metastasis than in those without [63.6% (21/33) vs 18.2% (2/11), chi2=5.132, P<0.01]. MSI was found in 8.9% (5/56) cases. AI was found in 26.8% (15/56) of all cases examined.</p><p><b>CONCLUSION</b>Frequent micro satellite alterations on chromosome 4 were existed in HCC. LOH, which represents tumor suppressor gene pathway, plays a more important role in hepatocarcinogenesis of HCC; MSI, representing mismatch repair gene pathway, arranges as the next.</p>

Relationship between microsatellite alterations on chromosome 8 and clinicopathological characteristics in hepatocellular carcinoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the features of microsatellite alterations and their association with clinicopathological characteristics of hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>Ten high-polymorphic microsatellite markers on chromosome 8 were selected to detect the loss of heterozygosity (LOH), microsatellite instability (MSI) and allelic imbalance (AI) in 56 HCCs using automatic capillary array electrophoresis DNA analysis system.</p><p><b>RESULTS</b>LOH was found in 37 of 56 HCCs (66.1%) on at least 10 locus. The three most frequently altered loci were D8S261 (53.5%, 23/43), D8S1721 (52.5%, 21/40) and D8S1771 (52.5%, 21/40). LOH on D8S277 was significantly higher in cases with positive serum HBsAg than in those with negative HBsAg (P < 0.01). Similarly, LOH on D8S261, D8S298 and D8S1733 occurred more frequently in patients with negative HBsAg than those with positive HBsAg (P < 0.01). LOH on D8S298 and D8S1771 were more frequent in tumors larger than 3 cm in size (P < 0.05 and P < 0.01 respectively). LOH frequencies of D8S1721 were significantly higher in cases with absent or partially encapsulated tumor than in those with intact tumor capsule (P < 0.05). LOH on D8S298 and D8S1771 were more frequently detected in tumors with intrahepatic metastasis than those without (P < 0.01). MSI was found in 12.5% (7/56) cases. AI was found in 19.6% (11/56) of all cases examined.</p><p><b>CONCLUSIONS</b>Microsatellite alterations on chromosome 8 were frequent in HCC. LOH, possibly representing alterations of the tumor suppressor pathway, may play an important role in hepatocarcinogenesis. MSI, reflecting a dysfunction of the mismatch repair pathway, may also contribute to this process, but in a less significant way. LOH at some particular loci is associated with certain clinicopathological parameters of human HCC.</p>

Morphologic variants and immunohistochemical features of hepatic angiomyolipoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the clinicopathological characteristics, immunohistochemical features and differential diagnosis of hepatic angiomyolipoma (AML).</p><p><b>METHODS</b>The clinicopathological features of hepatic AML were systematically examined in 44 surgically resected tumor specimens, with additional immunohistochemistry study using 10 relevant antibodies.</p><p><b>RESULTS</b>The tumors were composed of various amounts of three components, i.e. blood vessels, smooth muscle cells and adipose cells. According to the proportions of each of these tissue components, AML was subcategorized into the classical type (n = 13), myomatous type (n = 25), lipomatous type (n = 4), and angiomatous type (n = 2). Myoid cells displayed various morphology, including epithelioid, intermediate (ovoid or short spindle), spindle, oncocytic, and pleomorphic features. Hematopoietic elements were present as minor findings in eight tumors. Immunohistochemically, the tumor cells were strongly positive for HMB45 (44/44, 100%), SMA (38/38, 100%) and CD117 (30/38, 78.9%).</p><p><b>CONCLUSIONS</b>A correct diagnosis of hepatic AML might be difficult due to its various growth patterns and cell types. HMB-45 positivity in the myoid cells is a key feature for hepatic AML. CD117 may be another useful ancillary marker for reaching a definite diagnosis.</p>

Immunopathological features of hepatic angiomyolipoma: An analysis of 36 cases / 第二军医大学学报

To study the immunopathological characteristics and differential diagnosis of hepatic angiomyolipoma(AML).Methods:Thirty-six surgically resected hepatic AML were investigated clinicopathologically and immunohistochemically with 10 antibodies.Results:Hepatic AML occurred in 21 females and 15 males,with the mean age of 41.6 years(26-60 years old).The patients with AML often had no special symptoms even had large space-occupying lesions in the liver.The diameter of AML was 2.5 cm to 14 cm(mean 6.8 cm).Histologically,AML was composed of varying heterogeneous mixture of 3 tissue components:blood vessels,smooth muscle and adipose cells.Extramedullary hemopoiesis sometimes existed.According to tissue components,AML was subcategorized into mixed type(19.4%,n=7),lipomatous type(11.1%,n=4),myomatous type(66.7%,n=24),and angiomatous type(2.8%,n=1).The epithelioid smooth muscle cells were sensitive to HMB-45(100%),SMA(100%),and CD117(66.7%) staining.Conclusion:Hepatic AML often contains smooth muscle elements,which have varied morphological features and should be carefully differentiated from hepatocellular carcinoma,mesenchymal hamartoma，and tumors with rich fat or blood vessels.Immunohistochemical staining with HMB-45 and SMA are the best available markers for the diagnosis of hepatic AML.

Immunopathological features of hepatic angiomyolipoma: An analysis of 36 cases / 第二军医大学学报

To study the immunopathological characteristics and differential diagnosis of hepatic angiomyolipoma(AML).Methods:Thirty-six surgically resected hepatic AML were investigated clinicopathologically and immunohistochemically with 10 antibodies.Results:Hepatic AML occurred in 21 females and 15 males,with the mean age of 41.6 years(26-60 years old).The patients with AML often had no special symptoms even had large space-occupying lesions in the liver.The diameter of AML was 2.5 cm to 14 cm(mean 6.8 cm).Histologically,AML was composed of varying heterogeneous mixture of 3 tissue components:blood vessels,smooth muscle and adipose cells.Extramedullary hemopoiesis sometimes existed.According to tissue components,AML was subcategorized into mixed type(19.4%,n=7),lipomatous type(11.1%,n=4),myomatous type(66.7%,n=24),and angiomatous type(2.8%,n=1).The epithelioid smooth muscle cells were sensitive to HMB-45(100%),SMA(100%),and CD117(66.7%) staining.Conclusion:Hepatic AML often contains smooth muscle elements,which have varied morphological features and should be carefully differentiated from hepatocellular carcinoma,mesenchymal hamartoma，and tumors with rich fat or blood vessels.Immunohistochemical staining with HMB-45 and SMA are the best available markers for the diagnosis of hepatic AML.

Effects of nordihydroguaiaretic acid on genomic methylation pattern of malignant glioma cells and its significance / 第三军医大学学报

Objective　To investigate the genomic methylation pattern of a malignant glioma cell line in the process of differentiation induced by nordihydroguaiaretic acid (NDGA). Methods　Methylation-sensitive arbitrarily primed PCR was used to study the genomic methylation changes. Results　Fragments of genomic DNA of PCR products in control groups digested with MspⅠ were smaller than those with HpaⅡ. No large fragment could be identified and at least three fragments of different sizes were demonstrated in the control group. In NDGA treatment group, comparing MspⅠ digestion with HpaⅡ digestion, the amount of PCR products was smaller with more DNA bands. The amount of PCR products in NDGA treatment group was increased with more DNA bands compared with that in the control groups. Conclusion　The genomic methylation level in SHG-44 cells was increased by NDGA in the differentiation process of SHG-44 cells. It suggests that genomic methylation pattern may be one of the targets for glioma cell differentiation induced by NDGA.

Effects of nordihydroguaiaretic acid on genomic methylation pattern of malignant glioma cells and its significance / 第三军医大学学报

Objective　To investigate the genomic methylation pattern of a malignant glioma cell line in the process of differentiation induced by nordihydroguaiaretic acid (NDGA). Methods　Methylation-sensitive arbitrarily primed PCR was used to study the genomic methylation changes. Results　Fragments of genomic DNA of PCR products in control groups digested with MspⅠ were smaller than those with HpaⅡ. No large fragment could be identified and at least three fragments of different sizes were demonstrated in the control group. In NDGA treatment group, comparing MspⅠ digestion with HpaⅡ digestion, the amount of PCR products was smaller with more DNA bands. The amount of PCR products in NDGA treatment group was increased with more DNA bands compared with that in the control groups. Conclusion　The genomic methylation level in SHG-44 cells was increased by NDGA in the differentiation process of SHG-44 cells. It suggests that genomic methylation pattern may be one of the targets for glioma cell differentiation induced by NDGA.