Salivary Pepsin as an Intrinsic Marker for Diagnosis of Sub-types of Gastroesophageal Reflux Disease and Gastroesophageal Reflux Disease-related Disorders

Background/Aims@#To determine the value of salivary pepsin in discriminating sub-types of gastroesophageal reflux disease (GERD) and GERD-related disorders. @*Methods@#Overall, 322 patients with different sub-types of GERD and 45 healthy controls (HC) were studied. All patients took Gastroesophageal Reflux Disease Questionnaire (GerdQ) and underwent endoscopy and 24-hour esophageal pH monitoring and manometry. Salivary pepsin concentration (SPC) was detected by using colloidal gold double-antibody immunological sandwich assay. Oral esomeprazole treatment was administrated in the patients with non-erosive reflux disease (NERD) and extra-esophageal symptoms (EES). @*Results@#Compared to HC, patients with erosive esophagitis, NERD, EES, EES plus typical GERD symptoms, or Barrett’s esophagus had a higher prevalence of saliva and SPC (all P < 0.001). There was no significant difference in the positive rate for pepsin in patients with functional heartburn or GERD with anxiety and depression, compared to HC. After esomeprazole treatment, the positive rate and SPC were significantly reduced in NERD (both P < 0.001) and in EES (P = 0.001 and P = 0.002, respectively). Of the 64 NERD patients, 71.9% (n = 46) were positive for salivary pepsin, which was significantly higher than the rate (43.8%, n = 28) of pathological acid reflux as detected by 24-hour esophageal pH monitoring (P = 0.002). @*Conclusions@#Salivary pepsin has an important significance for the diagnosis of GERD and GERD-related disorders. Salivary pepsin and 24-hour esophageal pH monitoring may complement with each other to improve the diagnostic efficiency.

Catharmus tinctorius volatile oil promote the migration of mesenchymal stem cells via ROCK2/Myosin light chain signaling / 中国天然药物

MSC transplantation has been explored as a new clinical approach to stem cell-based therapies for bone diseases in regenerative medicine due to their osteogenic capability. However, only a small population of implanted MSC could successfully reach the injured areas. Therefore, enhancing MSC migration could be a beneficial strategy to improve the therapeutic potential of cell transplantation. Catharmus tinctorius volatile oil (CTVO) was found to facilitate MSC migration. Further exploration of the underlying molecular mechanism participating in the pro-migratory ability may provide a novel strategy to improve MSC transplantation efficacy. This study indicated that CTVO promotes MSC migration through enhancing ROCK2 mRNA and protein expressions. MSC migration induced by CTVO was blunted by ROCK2 inhibitor, which also decreased myosin light chain (MLC) phosphorylation. Meanwhile, the siRNA for ROCK2 inhibited the effect of CTVO on MSC migration ability and attenuated MLC phosphorylation, suggesting that CTVO may promote BMSC migration via the ROCK2/MLC signaling. Taken together, this study indicates that C. tinctorius volatile oil could enhance MSC migration via ROCK2/MLC signaling in vitro. C. tinctorius volatile oil-targeted therapy could be a beneficial strategy to improve the therapeutic potential of cell transplantation for bone diseases in regenerative medicine.

The clinical thinking of diagnosis and treatment of cryptogenic cirrhosis / 医学研究生学报

Cryptogenic cirrhosis (CC), cirrhosis of unknown cause, is common in clinic. With the improvement of the level of diagnosis and the diversification of the diagnosis and treatment, the incidence of CC is gradually declining. In the process of diagnosis and treatment, perfect clinical thinking can help clinicians to analyze and evaluate the existing clinical data, and it is expected to further improve the diagnostic rate of CC. In this paper, the clinical thinking of CC diagnosis and treatment is expounded from the aspects of disease spectrum thinking, anatomy thinking and macroscopic micro thinking.

Clinical characteristics of non-alcoholic fatty liver disease related hepatocellular carcinoma / 中国实用内科杂志

With the rising incidence, nonalcoholic fatty liver disease associated hepatocellular carcinoma(NAFLD-HCC) has become the main type of liver cancer. Therefore, to improve the level of clinical diagnosis and treatment of NAFLD-HCC is of great value. This paper summarized the clinical characteristics of NAFLD-HCC, and has found that the NAFLD-HCC has a special nature course,without typical clinical manifestation; it is common in male, often accompanied with the metabolic syndrome, with single tumor and good differentiation. Serology, CT imaging and pathology are important in diagnosis of NAFLD-HCC. Improve living habits and strengthen exercise can prevent the occurrence of NAFLD-HCC. Drug and surgical treatment is still the most important means to treat NAFLD-HCC, while the effects of those treatments used in NAFLD-HCC have no significant difference compared with that used in other types of liver cancer. To summarize clinical features will help improve the level of diagnosis and treatment of NAFLD-HCC.It's necessary to constantly explore the pathogenesis of NAFLD-HCC in order to formulate the individualized prevention strategy of NAFLD-HCC which is of great significance.

Effect of sinapine on H2O2-induced adipogenic differentiation of bone marrow mesenchymal stem cells / 中国病理生理杂志

AIM:To investigate the effects of sinapine,an effective monomer of Chinese medicine,on hydro-gen peroxide(H2O2)-induced adipogenic differentiation of bone marrow mesenchymal stem cells(BMSCs).METHODS:The undifferentiated rat BMSCs were identified and screened by flow cytometry.The adipogenic differentiation of BMSCs was induced by H2O2,and the toxicity of sinapine on BMSCs was tested by CCK-8 assay.After the modeling method and the concentration range of sinapine were determined,the lipid droplets in the cells were detected by Oil Red O semi-quanti-tative assay,and the optimal drug concentration was selected.Finally,Oil Red O assay was observed 24 h after drug inter-vention,and the expression of adipogenic differentiation-related proteins,adipocyte protein 2(aP2),peroxisome prolifera-tor-activated receptor γ(PPARγ)and glucose transporter 4(Glut4),at mRNA and protein levels in the BMSCs was deter-mined by qPCR and Western blot.RESULTS:Treatment with H2O2at 200 μmol/L for 1 h induced BMSCs to differentiate into adipocytes.Below the concentration of 40 μmol/L,sinapine had no toxicity to BMSCs.The best inhibitory concentra-tion of sinapine on adipogenic differentiation was at 15 μmol/L.The number of lipid droplets in sinapine(15 μmol/L) group was significantly lower than that in model group.In sinapine group,the expression of aP2,PPARγand Glut4 at mR-NA and protein levels was lower than that in model group(P<0.01).CONCLUSION: Sinapine inhibits H2O2-induced adipogenic differentiation of rat BMSCs.The mechanism may be related to the PPARγ/AMPK signaling pathway.

Ephrin-A2 and -A3 are negative regulators of the regenerative potential of Möller cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>In a previous study, we demonstrated that ephrin-A2 and -A3 negatively regulate the growth of neural progenitor cells in the central nervous system. Adult mice deficient in ephrin-A2 and -A3 (A2(-/-)A3(-/-)) displayed active ongoing neurogenesis throughout the brain, and mice deficient in ephrin-A3 alone showed increased proliferation of ciliary epithelium derived retinal stem cells. This study aimed to detect that the increase in proliferation and neurogenic potential of Müller cells is influenced by the absence of ephrin-A2 and -A3.</p><p><b>METHODS</b>We assessed the retinal and Müller cell expression of ephrin-As and their receptor and neural progenitor cell markers by immunostaining and real-time PCR. We cultured purified primary Müller cells derived from wild-type and A2(-/-)A3(-/-) mice in a defined culture medium that enables trans-differentiation of Müller cells into retinal neurons. To evaluate proliferating Müller cells in vivo, we injected 5'-ethylnyl-2'-deoxiuridine (EdU) intraperitoneally to adult mice.</p><p><b>RESULTS</b>Expression of ephrin-A2/A3 and their receptor EphA4 were detected in the retinas of adult mice, with EphA4 expression particularly enriched in Müller cells. Müller cells of A2(-/-)A3(-/-) mice exhibited significantly elevated expression of retinal progenitor cell markers, Pax6 and Chx10, when compared with those from wild-type mice. Moreover, a higher percentage of Müller cells of A2(-/-)A3(-/-) mice trans-differentiated and became recoverin+ and β-III-tublin+ in the culture than those from wild type mice. Strikingly, an increased number of EdU+ retinal cells was detected in the retinas of adult A2(-/-)A3(-/-) mice as compared with wild-type mice.</p><p><b>CONCLUSIONS</b>Ephrin-A2 and -A3 are negative regulators of the proliferative and neurogenic potentials of Müller cells. Manipulating ephrin-A signaling may thus represent a novel strategy for stimulating neuroregeneration from endogenous progenitors to participate in retinal repair in case of disease or damage.</p>

Intrinsic determinants of optic nerve regeneration / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To review the functions of these intracellular signals in their regulation of retinal ganglion cell (RGC) axon regeneration.</p><p><b>DATA SOURCES</b>Relevant articles published in English or Chinese from 1970 to present were selected from PubMed. Searches were made using the terms "intrinsic determinants, axon regeneration, RGC, optic nerve regeneration, and central nervous system axon regeneration."</p><p><b>STUDY SELECTION</b>Articles studying the mechanisms controlling RGC and central nervous system (CNS) axon regeneration were reviewed. Articles focusing on the intrinsic determinants of axon regeneration were selected.</p><p><b>RESULTS</b>Like other CNS neurons of mammals, RGCs undergo a developmental loss in their ability to grow axons as they mature, which is a critical contributing factor to the failure of nerve regeneration and repair after injury. This growth failure can be attributed, at least in part, by the induction of molecular programs preventing cellular overgrowth and termination of axonal growth upon maturation. Key intracellular signals and transcription factors, including B cell lymphoma/leukemia 2, cyclic adenine monophosphate, mammalian target of rapamycin, and Krüppel-like transcription factors, have been identified to play central roles in this process.</p><p><b>CONCLUSIONS</b>Intense effort and substantial progress have been made to identify the various intrinsic growth pathways that regulate RGC axon regeneration. More work is needed to elucidate the mechanisms of and the interrelationship between the actions of these factors and to successfully achieve regeneration and repair of the severed RGC axons.</p>

Insulin-like growth factor-1-mediated paracrine pro-proliferative effect of bone marrow mesenchymal stem cells on injured L02 hepatocytes / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To explore the therapeutic effect of bone marrow mesenchymal stem cells (BM-MSCs) on injured hepatocytes mediated by paracrine mechanisms and to investigate the potential molecular mechanism of this action.</p><p><b>METHODS</b>A contact-independent model of aberrant hepatic microenvironment was established by co-culturing BM-MSCs with D-galactosamine (D-GalN)-injured human L02 hepatic cells using a transwell assay platform. Secreted levels of insulin-like growth factor-1 (IGF-1) were measured by enzyme-linked immunosorbent assay of the co-culture supernatant. Expression of the IGF-1 receptor (IGF-1R) was assessed by Western blot. The effect of exogenous IGF-1 on proliferation of D-GalN-injured L02 cells was examined by MTT assay.</p><p><b>RESULTS</b>Upon co-culture, BM-MSCs promoted proliferation of D-GalN-injured L02 cells in a contact-independent manner (absorbance values of at 24 h: 0.36+/-0.08, 48 h: 0.52+/-0.06, and 96 h: 0.68+/-0.06; vs. uninjured cells t = 2.493, 3.116, and 2.285, respectively; all P less than 0.05). Robust expression of IGF-1 was identified in the supernatants of co-cultures and was demonstrated to have been secreted mainly from BM-MSCs under the influence of D-GalN-injured L02 cells. Constitutive expression of IGF-1R was found in the D-GalN-injured L02 cells and blocking of IGF-1R by a neutralizing antibody significantly inhibited the paracrine pro-proliferative effect of co-cultured BM-MSCs at 24 h, 48 h, and 72 h (t = 2.909, 2.328, and 2.560, respectively; all P less than 0.05).</p><p><b>CONCLUSION</b>BM-MSC-derived IGF-1 plays an important role in the paracrine pro-proliferative effect on D-GalN-injured L02 hepatocytes by engaging with the constitutively expressed IGF-1R on L02 cells.</p>

SREBP-1c knockdown attenuated fatty degeneration in hepatic L02 cells and inhibited CCL2 and FGF21 protein expression / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the effect of SREBP-1c silencing on lipid metabolism and expression of inflammatory chemokines in a NAFLD model with endoplasmic reticulum stress.</p><p><b>METHOD</b>NAFLD model was established in L02 cells treated with oleic acid. SREBP-1c expression was inhibited using RNA interference with a p Silencer-1.0-U6-4476 vector. After transfection with p Silencer-1.0-U6-4476 or control vector for 0 h, 24 h, 48 h and 72 h, the extent of fatty degeneration was shown by Oil Red O staining. The mRNA and protein expression of inflammatory chemokine CCL2 and basic fibroblast growth factor-21 (FGF21) were determined by real time PCR and Western blot respectively.</p><p><b>RESULTS</b>SREBP-1c silenced L02 cells showed fat droplets with smaller diameter and attenuated fatty deposition, as compared with control cells. The relative CCL2 mRNA levels in SREBP-1c silencing vector transfected L02 cells were 1.03+/-0.11 for 0 h, 1.11+/-0.21 for 24 h, 0.88+/-0.16 for 48 h, and 1.05+/-0.15 for 72 h, which showed no significant difference as compared with control cells (P>0.05, respectively). In addition, no difference was found between the different time points within the same group (P>0.05). However, CCL2 protein levels in SREBP-1c silenced cells were 1.19+/-0.15, 1.07+/-0.18, 0.48+/-0.14, and 0.05+/-0.24 after transfection for 0 h, 24 h, 48 h, and 72 h respectively, which were significantly downregulated as compared to the control group (P<0.01). And CCL2 protein levels between different time points in SREBP-1c silenced cells were also distinct (P<0.01). The relative FGF21 mRNA levels in SREBP-1c silenced L-02 cells were 1.01+/-0.08, 0.91+/-0.22, 0.98+/-0.20, and 1.02+/-0.12 for 0 h, 24 h, 48 h, and 72 h respectively, which were not statistically different as compared with the corresponding control cells. Statistic difference of FGF21 mRNA levels in SREBP-1c knockdown cells of different time points was not found (P>0.05). In striking contrast, robust down regulation of FGF21 protein in SREBP-1c silenced cells was observed, with 0.81+/-0.05, 0.66+/-0.12, 0.58+/-0.08 and 0.19+/-0.13 after transfection for 0 h, 24 h, 48 h and 72 h respectively, as compared to control group (P<0.01). And differences in FGF21 protein level between different time points in SREBP-1c silenced cells were also demonstrated (P<0.01).</p><p><b>CONCLUSION</b>SREBP-1c knockdown attenuated fatty deposition in oleic acid treated L02 cells. In addition, silencing of SREBP-1c expression reduced expressions of CCL2 and FGF21 proteins posttranscriptionally, which may play a role in endoplasmic reticulum stress induced inflammatory response in NAFLD.</p>

Therapeutic effect of triple regulated adenovirus vector expressing mda7/IL24 on liver cancer in vitro / 肿瘤

Objective: To construct an E1 A-deleted 24-bp triple regulated replicative adenovirus vector SG600/interleukin24 (IL24), which was driven by both hTERT promoter and HRE promoter. The level of IL24 in liver cancer cells was determined and the replication capacity of SG600/ IL24 and its killing effects on liver cancer cells were observed. Methods: SG600-IL24 vector was constructed using DNA cloning and recombination techniques. The IL24 gene expression in liver cancer cell lines SMMC-7721 and BEL-7404 and normal cell line BJ was detected by ELISA assay. The replications of SG600/IL-24 in different cell lines were determined by evaluating TCID50 (50% tissue culture infectious dose) at 49 and 96 h. In vitro cell-killing effects of SG600/IL24 on the three liver cancer cell lines were analyzed by MTT assay and CPE (cytopathic effect) staining method at different MOI values. Results: IL24 was over-expressed in both SMMC-7721 and BEL-7404 cells but was weakly expressed in BJ cells. At 48 and 96 h post infection the replication of SG600/IL-24 were 794 and 7940 folds in SMMC-7721 cells; 622 and 7 810 folds in BEL-7404 cells; 20 and 200 folds in BJ cells. MTT assay showed that the MOI values of SG600/IL24 for killing 50% and 90% cells were 0.3 and 5 for SMMC-7721 cells; 3 and 20 for BEL-7404 cells; 50 and 150 for BJ cells. CPE staining demonstrated that SG600/IL24 had significant killing effects on both liver cancer cells SMMC-7721 and BEL-7404 but had no significant influence on BJ cells. The cell-killing capability of SG600/IL24 was superior than that of replicative adenovirus ZD55/IL24 and non replicative adenovirus Ad-IL24. Conclusion: After SMMC-7721 and BEL-7404 liver cancer cells are infected with SG600/1124 at high efficiency, the virus replication is active and the expression of IL24 increases greatly. SG600/IL24 has specific cell-killing effects on the two liver cancer cell lines but has no significant influence on normal cells. This study provides a basis for further investigating the effect of SG600/IL24 on liver cancer in vivo.

The expression and significance of osteopontin in the development of nonalcoholic fatty liver fibrosis in rats / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the changes of osteopontin (OPN) in the liver tissues during nonalcoholic fatty liver fibrosis in rats and to explore the effect of OPN in the development of nonalcoholic fatty liver fibrosis.</p><p><b>METHODS</b>Fifty-six male Wistar rats were randomly divided into a control group (8 rats) and a high-fat diet group. The high-fat diet group was divided into 6 subgroups (8 rats in each subgroup) with high-fat feedings for 4, 8, 12, 16, 20 or 24 weeks. Conventional histochemical, HE, Masson-trichrome and immunohistochemical staining for alpha-smooth muscle actin (a-SMA) were performed with the liver histological preparations. The expression of OPN was detected with reverse transcription and polymerase chain reactions and Western blot.</p><p><b>RESULTS</b>Levels of OPN in liver tissues in rat nonalcoholic fatty liver fibrosis induced by high-fat diet were significantly increased over those in the control group (F=7.15, P less than 0.01). OPN expressions were closely correlated with a-SMA and nonalcoholic fatty liver fibrosis, and correlation coefficients of the two groups were 0.94 and 0.82, and both P values were less than 0.01.</p><p><b>CONCLUSION</b>Expression of OPN increases dramatically in the livers during the development of nonalcoholic fatty liver fibrosis, and OPN may play an important role in this event.</p>

Histological changes of an injectable rhBMP-2/calcium phosphate cement in vertebroplasty of rhesus monkey / 中华外科杂志

<p><b>OBJECTIVE</b>The histological changes of rhBMP-2/calcium phosphate cement (CPC) were evaluated in vertebroplasty on nonhuman primate models in order to determine the feasibility of this carrier formulation instead of PMMA.</p><p><b>METHODS</b>Percutaneous vertebroplasty (PVP) was performed in 4 adult rhesus monkeys which were evenly distributed in two groups. Ten vertebral bodies(VBs) from T10 to L7, of each rhesus were selected, and the 20 VBs in each group were randomly divided into 3 sub-groups. Group A:8 VBs, filled with rhBMP-2/CPC; Group B:6 VBs, filled with injectable PMMA; Group C:6 VBs, as control, filled with normal saline. The 2 rhesus monkeys in each group were killed at 2 and 6 months after operation, respectively, and the specimens of all the 40 VBs were collected for histological examination.</p><p><b>RESULTS</b>In group A,radiographic and histologic studies confirmed that part of the rhBMP-2/CPC cement degraded with new bone and new vessels ingrowth into the material after 2 months. No gap, fibrous hyperplasia or sclerotic callus was found in the interface. After six months, the cement was almost completely replaced by mature bone tissue. In group B, no new bone formation and material degradation but inflammatory cell infiltration and fibrous membrane gap were found 2 months after operation. After 6 months, the inflammatory cell infiltration subsided, the fibrous membrane gap became narrower, but there were still no new bone formation and material degradation. In group C, the tunnels were filled with irregular new trabeculae after 2 months and unrecognizable from the surrounding mature bone after 6 months, indicating the completion of bone healing.</p><p><b>CONCLUSIONS</b>With the characteristic of osteo-induction, the rhBMP-2/CPC can accelerate the healing of vertebral bone in nonhuman primates. Bone substitution is synchronous with material degradation, and the complete degradation of this material in late stage can avoid the potential adverse effects of PMMA on contiguous vertebral fracture and annulus degeneration. It might be a perfect bone substitute material for vertebroplasty instead of PMMA in the future.</p>

Effects of rosiglitazone on Kruppul-like factor 6 (KLF6) signaling in the livers of rats with nonalcoholic fatty liver fibrosis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the effects of rosiglitazone on Kruppule-like factor 6 (KLF6) and its target gene TGFâ1 during the development of nonalcoholic fatty liver fibrosis.</p><p><b>METHOD</b>Thirty-six Wistar rats were divided into three groups: a control group, a high fat model group and an intervention group. Their blood serum TG, FFA, AST, ALT, HA, LN and CIV were measured. Hepatic expressions of KLF6, TGFbeta1 and alpha-SMA were detected by RT-PCR and immunohistochemistry. The pathological features and the degree of liver fibrosis before and after the rosiglitazone intervention were also studied.</p><p><b>RESULTS</b>The contents of TG, FFA, AST, ALT, HA, LN and CIV, the expression of KLF6, TGFbeta1 and alpha-SMA mRNA, and the degree (score) of liver fibrosis at the 24th week in the model group were significantly higher than those in the control group (P<0.01) but they were lower in the rosiglitazone intervention group (P<0.05). The protein expression of a-SMA was also lower in the intervention group compared with that of the model group.</p><p><b>CONCLUSION</b>Rosiglitazone, to a certain extent, can inhibit KLF6-TGFbeta1 signal transduction by inducing expression of PPAR-gamma, and then inhibit the activation of hepatic stellate cells and minimize hepatic fibrosis.</p>

Effects of TNF alpha on the expression of SCAP and triglyceride contents in cultured steatotic hepatocytes / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To explore the effects of TNF alpha on the expression of sterol regulatory element binding proteins cleavage activating protein (SCAP) and triglyceride contents in cells of a model of cultured steatotic hepatocytes.</p><p><b>METHODS</b>A steatotic hepatocytes model was established by treating L-02 cell strain with oleic acid. The cells were treated with TNF alpha and/or TNF alpha antibody. The cells were divided into six groups: a control group (C), a model group (F), a control group with TNF alpha (C1), a control group with TNFalpha antibody (C2), a model group with TNFalpha(F1) and a model group with TNFalpha antibody (F2). The expression of SREBP-1c mRNA was measured with RT-PCR; the protein expression of SCAP was measured by Western blot; lipid droplets in the hepatocytes were observed with oil red O staining; the contents of triglyceride in hepatocytes were measured with an analytical kit.</p><p><b>RESULTS</b>The mRNA expression of SCAP in the groups treated with TNF alpha were upregulated compared with those of the control group (C1 vs C increased 67%, F1 vs F increased 55%, F = 212.98), the protein expression of SCAP in the groups treated with TNF alpha was upregulated compared with those of the control group (C1 vs C increased 45%, F1 vs F increased 95%, F = 104.3), and triglyceride contents in hepatocytes of these groups were increased compared with those of the control group [C (2.02+/-0.67) mg/10(7) cells, F(7.79+/-1.35) mg/10(7) cells, F1(13.36+/-1.99) mg/10(7) cells, F = 82.94].</p><p><b>CONCLUSION</b>TNF alpha upregulates the expression of SCAP and promotes the synthesis of triglyceride; it probably participates in the process of developing steatosis of hepatocytes.</p>

The relationship between the opening of mitochondrial permeability transition pores of cultured hepatocytes with their apoptoses in a non-alcoholic fatty liver disease model / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the opening of the mitochondrial permeability transition pores of the cultured hepatocytes in a non-alcoholic fatty liver disease model and its relationship with apoptosis of the cells.</p><p><b>METHODS</b>Oleic acid was used to induce cultured L02 hepatocyte steatotic in making a model of NAFLD. The steatotic hepatocytes were detected with oil red O staining; the opening of the mitochondrial permeability transition pores was observed under a fluorescence microscope. The apoptosis of the cells was detected with a flow cytometer.</p><p><b>RESULTS</b>After adding oleic acid to the cultured hepatocytes, a model of steatosis of human hepatocytes was established after 24 hours. Oleic acid opened the mitochondrial permeability transition pores of the L02 hepatocytes (72.58%+/-2.78%) more than that in the control group (8.28%+/-4.98%) and the difference was statistically significant (P < 0.01). Apoptosis index of the steatotic hepatocytes at 24 hours and 48 hours were 11.09%+/-4.95% and 15.24%+/-2.45%. They were also higher than those of the control group (4.56%+/-1.25%) (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>Opening the mitochondrial permeability transition pores may be the basis of the apoptosis of steatotic hepatocytes in vitro, and it also may be related to the steatosis of NAFLD in human beings.</p>

The significance and effects of liver X receptor alpha in nonalcoholic fatty liver disease in rats / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To explore liver X receptor alpha (LXR alpha) gene changes and their significance in nonalcoholic fatty liver disease (NAFLD) in rats.</p><p><b>METHODS</b>A rat model of nonalcoholic fatty liver disease was produced with a fatty diet regime (feeding group, FG). Rats fed with normal diet served as controls (CG). The mRNA and protein expressions of LXR alpha in liver tissues were detected by reverse transcription and polymerase chain reaction (RT-PCR) and Western blot.</p><p><b>RESULTS</b>The concentration of free fatty acid (FFA) in the sera of GF rats started to increase to 0.33 mmol/L after 4 weeks of fat diet feeding, while the FFA of the CG was just 0.24+/-0.03 mmol/L, and the difference was significant (P<0.05). The concentration of ALT and AST in sera of the FG rats started to increase to 75.8 U/L and 138.9 U/L at the 8th week, much higher than those of the CG (P<0.01), and at the 12th week they increased further (P<0.01). Meanwhile, the mRNA and protein expressions of LXR alpha at the 2nd week was significantly increased to 0.62 (P>0.01) and its peak was reached at the 12th week (P<0.01). There was a significant positive correlation between the expression of LXR alpha and the degree of NAFLD.</p><p><b>CONCLUSION</b>The changes of LXR alpha gene are closely related to the development of nonalcoholic fatty liver disease.</p>

Effect of fatty acids from plastrum testudinis on proliferation of rat bone mesenchymal stem cell / 生物工程学报

To investigate the components in Plastrum Testudinis which have effects on the proliferation of rat bone marrow mesenchymal stem cells( bMMSCs), the active parts of plastrum testudinis which can promote proliferation of rat mesenchymal stem cells were extracted by petroleum aether. The activities of inducing the proliferation of bMMSCs were studied by MTT assay and flow cytometry. The chemical components of extraction were analyzed by GC-MS. The results showed that the petroleum aether extraction can obviously promote the proliferation of the stem cells. The main components are long-chain fatty acids, cholesterols and cholest-4-en-3-one, and palmitic acid, stearic acid and cholest-4-en-3-one have effects on proliferation of bMMSCs. In plastrum testudinis, fatty acids can promote the proliferation of bMMSCs but not increase overly. This provide the experiment basis, and offer important reference for Traditional Chinese Medicine that researches stem cells.

Coexpression of hepatitis B virus X gene and hepatitis C virus C gene in HepG2 cells and its effect on the expression of VEGF / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To establish an experimental model of HCV C-HBV X co-expression protein and explore its effect on the expression of VEGF.</p><p><b>METHODS</b>The HBV X gene was recovered by enzyme excision and inserted into PBK-CMV and PBK-HCVC, and recombinant plasmids PBK-X and PBK-X-C were constructed. The plasmids PBK-CMV, PBK-X, PBK-HCVC and PBK-X-C were transfected into HepG2 cells with liposomes. After being selected by G418, resistant colonies were obtained. Reverse transcription PCR and Western blot were used to show HBV X and HCV core protein expression. VEGF was analyzed using immunohistochemical methods and Western blot.</p><p><b>RESULTS</b>The recombinant plasmid PBK-X-C expressed HBV X and HCV core protein efficiently under the control of the vectors promoter. VEGF and VEGF mRNA of the cells co-expressing HCV C-HBV X proteins were higher than those cells expressing HBV X, HCV C and vector alone.</p><p><b>CONCLUSION</b>HBV X-HCV C co-expression protein can increase the expression of VEGF of HepG2 cells. It suggests that HBV and HCV have a synergic action in the carcinogenesis.</p>

Liver regenerative capacity after partial hepatectomy in rats with nonalcoholic fatty liver disease / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To explore the changes of liver regeneration after partial hepatectomy on rats with nonalcoholic fatty liver disease (NAFLD) caused by a high fat diet.</p><p><b>METHODS</b>One hundred Wistar rats were randomly divided into a control group (group C, n = 45), fed with normal diet, and a NAFLD group, fed with fat-rich diet (group F, n = 55). All rats had a 70% partial hepatectomy at the end of the 12th week. They were sacrificed at postoperative 0, 1, 12, 24, or 36 hours and the percentages of their regenerated liver masses were calculated. The mitosis index was measured microscopically and the changes of cell ultrastructure were observed under an electron microscope. The expression of proliferating cell nuclear antigen (PCNA) was detected using immunohistochemistry. The expression of mRNA of cyclin D1 was measured by RT-PCR.</p><p><b>RESULTS</b>The light and electron microscopy showed that the hepatic sinusoids expanded at an early period after the partial hepatectomy. The mitochondria and rough endoplasmic reticulum (RER) expanded and their number increased. The mitosis index was increased. The sinusoids of the livers in group F were narrow and irregular. The nuclei were smaller and the necrotic cells increased. As compared with the control group, the mitosis index was significantly decreased (P < 0.01), and the regenerative liver weight ratio in group F was lower at postoperative 12 h, 24 h, 36 h (P < 0.01). PCNA labeling index in group F also was lower than that in group C. The peak of the PCNA in group F was later than that of the control group (P < 0.01). In group C, the mRNA of cyclin D1 peaked at the 24th hour after the partial hepatectomy, and then decreased afterwards. In group F, it was lower than that of group C at the same time (P < 0.01).</p><p><b>CONCLUSION</b>After NAFLD rats had partial hepatectomies, the capacity of their liver regeneration decreased and the peak of DNA synthesis was delayed, and at the same time the morbidity of the rats increased.</p>

The expression and the significance of L-FABP and FATP4 in the development of nonalcoholic fatty liver disease in rats / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the effect of liver fatty acid binding protein(L-FABP) and fatty acid transport protein (FATP4) in the development of nonalcoholic fatty liver disease (NAFLD) in rats.</p><p><b>METHODS</b>The expression of L-FABP and FATP4 genes was examined in fatty liver rats by reverse transcription and polymerase chain reaction amplification and Western blot methods.</p><p><b>RESULTS</b>In the high fat diet group (F), mRNA and protein expression of L-FABP and FATP4 were increased at 2 weeks, and they increased remarkably at 12 weeks (P < 0.05; L-FABP mRNA F=124.9, protein expression F=92.6; FATP4 mRNA F=602.9, protein expression F=108.8).</p><p><b>CONCLUSION</b>The high expression of L-FABP and FATP4 at the early stage is an adaptive reaction of the body, With the advanced expression of the L-FABP and FATP4, it can lead to a fatty acid disequilibrium and then result in nonalcoholic fatty liver disease in the rats.</p>