Development of membrane anatomy theory in gastric cancer surgery / 中华胃肠外科杂志

In the past decade, the concept of membrane anatomy has been gradually applied in gastric cancer surgery. Based on this theory, D2 lymphadenectomy plus complete mesogastric excision (D2+CME) has been proposed, which has been demonstrated to significantly reduce intraoperative bleeding and intraperitoneal free cancer cells during surgery, decrease surgical complications, and improve survival. These results indicate that membrane anatomy is feasible and efficacious in gastric cancer surgery. In this review, we will describe the important contents of membrane anatomy, including "Metastasis V"(2013, 2015), proximal segmentation of dorsal mesogastrium (2015), D2+CME procedure (2016), "cancer leak"(2018), and surgical outcomes of D2+CME (2022).

A decade's review for membrane anatomy: the setting,events in it, order formed by primary fascia and serous membrane / 中华胃肠外科杂志

The successful report of total mesorectal excision (TME)/complete mesocolic excision (CME) has encouraged people to apply this concept beyond colorectal surgery. However, the negative results of the JCOG1001 trial denied the effect of complete resection of the "mesogastrium" including the greater omentum on the oncological survival of gastric cancer patients. People even believe that the mesentery is unique in the intestine, because they have a vague understanding of the structure of the mesentery. The discovery of proximal segment of the dorsal mesogastrium (PSDM) proved that the greater omentum is not the mesogastrium, and further revised the structure (definition) of the mesentery and revealed its container characteristics, i.e. the mesentery is an envelope-like structure, which is formed by the primary fascia (and serosa) that enclose the tissue/organ/system and its feeding structures, leading to and suspended on the posterior wall of the body. Breakdown of this structure leads to the simultaneous reduction of surgical and oncological effects of surgery. People quickly realized the universality of this structure and causality which cannot be matched by the existing theories of organ anatomy and vascular anatomy, so a new theory and surgical map- membrane anatomy began to form, which led to radical surgery upgraded from histological en bloc resection to anatomic en bloc resection.

Demonstration and significance on proximal segment of dorsal mesogastrium / 中华胃肠外科杂志

In radical gastrectomy, D2 systemic lymphadenectomy, which includes complete resection of the bursa sac and omentum, and D2 extended lymphadenectomy outside the bursa sac, is a standard procedure accepted by gastrointestinal surgeons generally. However, a series of clinical trials showed that both D2 extended lymphadenectomy and bursectomy could not improve oncologic benefit, but increase surgical risk. These findings showed a lot of conflicts in gastric cancer surgery, gastrointestinal surgery, even in oncological surgery. It was demonstrated that bursa sac and greater omentum were neither mesogastrium nor the proximal segment of dorsal mesogastrium (PSDM), which has been identified recently. Local physiological structures (such as blood vessels and lymphatic nodes) and pathological events (such as lymph nodes metastasis and metastasis V) only occur in mesentery in broad sense (i.e. PSDM). Broken PSDM during radical gastrectomy can result in cancer cell leakage into the operational field. Therefore, complete PSDM excision in the D2 field (D2+CME) is suggested as a better procedure for local advanced gastric cancer, which can get benefits not only in surgical hazard, but also in oncologic result. The results of PSDM research could lead to three changes: (1) resolving some long standing problems in gastric cancer surgery, gastrointestinal surgery, and even oncologic surgery; (2) opening an new era for finding and utilizing extra-intestinal mesentery in broad sense; (3) formulating the theory of membrane anatomy which may update, iterate and upgrade related information of classical anatomy, pathology, surgery and oncology.

Scientific paradigm of membrane anatomy / 中华胃肠外科杂志

Anatomical plane and fascia have been described in medical behaviors for hundreds of years since the appearance of anatomy and operation. Generally, these descriptions can be sorted into three theories, i.e. plane surgery, fascia theory and mesentery anatomy. However, these theories are difficult to satisfy the scientific paradigm that includes consistency in description, independence in validation, potential to solve practical problems, and the interaction of the above-mentioned theries. Recently, membrane anatomy was proposed as the anatomy of mesentery and its beds in broad sense. Behind it lies fascia membrane/serous membrane structure, as well as inherent life events and general order. Mesentery in broad sense is described as the fascia membrane/serous membrane in serous cavity, which envelops and suspends the organ/tissue and its feeding structures to the posterior wall of the body. Anatomy is the setting/structure, in which life events/functions occur. In the research and discussion of membrane anatomy, abiding by the scientific paradigm and upholding the scientific spirit are the only way to obtain reliable knowledge and the criterion for in-depth scientific research.

Simultaneous Determination of Seven Components in Blossoms of Inula nervosa by UHPLC / 中国实验方剂学杂志

Objective::To establish an ultra-high-performance liquid chromatography (UHPLC) method for simultaneous determination of seven components(chlorogenic acid, 3, 5-dicaffeoylquinic acid, 1, 5-dicaffeoylquinic acid, kaempferol-3-O-rutinoside, quercetin, kaempferol and thymol) in blossoms of Inula nervosa, and provide references for its quality control. Method::The separation was performed on an Agilent Poroshell 120 C18 column (3.0 mm×100 mm, 2.7 μm) with 0.1% formic acid(A) and methyl (B)as mobile phase for gradient elution(0-4 min, 2%B; 4-6 min, 2%-5%B; 6-10 min, 5%-10%B; 10-20 min, 10%-20%B; 20-30 min, 20%-27%B; 30-37 min, 27%-25%B, 37-45 min, 25%-32%B; 45-68 min, 32%-58%B; 68-75 min, 58%-25%B; 75-82 min, 25%-2%B; 82-90 min, 2%B). The flow rate was 0.3 mL·min-1 and the detection wavelength was 254 nm. Result::There was a good linear relationship between the concentration and peak area of all the seven components in the investigated concentration range (r>0.999). The average recoveries ranged from 97.80% to 101.28% with RSD≤3.0%. Cluster analysis of SPSS software and principal component analysis of SIMCA software can be used to intuitively classify samples from four different origins. Conclusion::The established method is simple and fast with high precision, which can be used to compare the differences of blossoms of Inula nervosa from different origins and efficiently control its quality.

Effects of PPARγ on the cholesterol efflux of C57 mice peritoneal macrophage in inflammatory conditions / 中国现代普通外科进展

Objective:To explore the effects of PPARγ on the cholesterol efflux of C57 mice peritoneal macrophage.Methods:Firstly constructing C57 mice model under different metabolic situation including high-fat diet and acute inflammation then isolate and culture its peritoneal macrophage,observing the expressions of PPARγ and IκB-α,identify the character of macrophage cholesterol efflux in every group.Then pretreat the normol C57 mice peritoneal macrophage with PPARγ ligand ciglitazone and PPARγ antisense oligonucleotide,observing the effect to cholesterol efflux after simulated with LPS in vitro.Results:The level of mice serum lipids of high fat diet group was significantly higher than that of normal diet group.The results of Western-blotting showed that the expression of PPARγ protein in groups of HFD and stimulated by LPS were significantly higher than that of control group.The expression in groups stimulated by LPS was all lower significantly than in control grouph.The determination of cholesterol efflux showed that this function of macrophage with HFD was more enhanced than that of control group but was inhibited in group stimulated by LPS.To normol peritoneal macrophage pretreat with PPARγ antisense oligonucleotide and stimulated by LPS,the expression of PPARγ protein was lower than that of control group but the expression of IκB-α was depressed obviously.Conclusion:The PPARγ ligand ciglitazone can increase the cholesterol efflux of C57 mice peritoneal macrophage and weaken the inhibition stimulated by LPS.The PPARγ antisense oligonucleotide can depress it and aggravate the inhibition.

Treatment strategy for hepatic trauma / 中华创伤杂志(英文版)

Liver is one of the organs with the highest injury rate, and in recent decades, the guidelines for the treatment of liver trauma have changed considerably. Now, there is a growing consensus that the most important step is diagnosis and depending upon the degree of severity, non-operative therapy is the main treatment method for hepatic trauma if conditions permit. For serious hepatic trauma patients such as those with hemodynamic instability, they should be operated upon as soon as possible. Regardless of the surgical options, doctors should control damage to patients and try to prevent complications. New therapies such as hepatic artery embolization and liver transplantation have been more and more used for the treatment of serious hepatic damage in clinics.

Over-expression of receptor-interacting protein 140 in tumor-associated macrophages suppresses invasion and proliferation of hepatoma cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the role of receptor-interacting protein 140 (RIP140) in tumor-associated macrophages (TAMs) in the invasion and proliferation of hepatoma cells in vitro.</p><p><b>METHODS</b>Western blotting, qRT-PCR and flow cytometry were performed to examine the effects of lentivirus-mediated RIP140 over-expression in mouse peritoneal macrophages (PMs). Western blotting, qRT-PCR and immunofluorescence staining were used to detect the expression of RIP140 in TAMs following stimulation of the PMs with hepatocellular carcinoma conditioned medium (HCM) for 24 h. The polarization index and the expression of NF-κB and IL-6 were detected using qRT-PCR in TAMs in HCM-stimulated PMs with or without RIP140 over-expression. Transwell assay and flow cytometry were used to estimate the cell invasion and apoptosis. HE staining and immunohistochemical staining were used to analyze the effects of RIP140-over-expressing macrophages on the growth and tumor formation of H22 cells in BALB/c nude mice.</p><p><b>RESULTS</b>The lentivirus vector efficiently mediated RIP140 over-expression in mouse PMs. HCM stimulation significantly inhibited RIP140 expression in the TAMs and promoted their M2-like polarization. Over-expression of RIP140 in PMs suppressed the invasion and induced apoptosis of HCC cells. RIP140 over-expression inhibited HCM-induced M2 polarization and the activation of NF-κB/IL-6 axis in the TAMs, and RIP140- overexpressing TAMs obviously suppressed the growth of H22 cell xenograft in nude mice.</p><p><b>CONCLUSION</b>Over-expression of RIP140 in TAMs suppresses the growth and proliferation of hepatoma cells possibly by inhibiting M2 polarization of the TAMs.</p>

Effect of inhibiting TIM-4 function in Kupffer cells on liver graft rejection in mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of inhibiting TIM-4 function in Kupffer cells (KCs) on liver graft rejection in mice and explore the underlying mechanism.</p><p><b>METHODS</b>Mouse models of orthotopic liver transplantation were treated with a control mAb group and TIM-4 mAb. The activated KCs were assayed with immunohistochemistry after operation. The expression of TIM-4 in KCs were assayed with Western blotting and RT-PCR and the levels of AST, ALT, TBIL, TNF-α, IFN-γ and CCL2 were assayed detected. The expression of TIM-4 in KCs was observed with laser confocal microscopy. HE staining was used to observe the microstructure of the liver tissues, and the number of CD25Foxp3T cells was determined using with flow cytometry; the proteins levels of p-P65and p-P38 were assayed with Western blotting. The donor mice were treated with clodronate liposomes to destroy the KCs in the liver before transplantation, and the liver grafts were examined for graft rejection.</p><p><b>RESULTS</b>The number of activated KCs in the liver graft increased progressively over time. Compared with the sham-operated group, the liver graft showed significantly increased TIM-4 protein and mRNA levels at 1, 3, and 7 days after transplantation (P<0.05) and increased levels of AST, ALT, TBIL, TNF-α, IFN-γ and CCL2 at 7 days (P<0.05). The graft in TIM-4 mAb group showed mild pathological changes with a mean RAI score of 2.67∓0.75, which was significantly lower than that in control mAb group (P<0.05). The mean survival time of the recipient mice was 53.8∓6.4 days in TIM-4 mAb group, significantly longer than that in the control mAB group (14.5∓2.9 days, P<0.05). Donor treatment with clodronate liposomes resulted in comparable RAI scores in TIM-4 mAb and control mAb groups (8.01∓0.64 vs 7.93∓0.56, P>0.05). The protein levels of p-P65 and p-P38 in TIM-4 mAb group were significantly lower than those in control mAb group (P<0.05), and CD25Foxp3T cells in the liver graft increased significantly in TIM-4 mAb group.</p><p><b>CONCLUSION</b>Inhibition of TIM-4 function in KCs reduces the production of inflammatory factors after liver transplantation possibly by inhibiting the NF-κB and MAPK signaling pathways and promoting the proliferation of Foxp3Treg cells to induce allograft tolerance.</p>

Choledochoduodenal fistula in Mainland China: a review of epidemiology, etiology, diagnosis and management

PURPOSE: Choledochoduodenal fistula (CDF) is an extremely rare condition even in the most populous nations. However, diagnostic tools are inadequate for the young surgeon to be made aware of such a rare condition before surgery. Hence, basic understanding of the epidemiology, etiology, and management for this unusual but discoverable condition are necessary and essential. METHODS: The exclusive case reports of CDF, which were published from 1983 to 2014 concerning mainland Chinese people, were performed to review the epidemiology, etiology, and management. RESULTS: A total of 728 cases were incorporated into this review among 48 papers. More than half of the CDF cases were female (416) with an average age of 57.3 years. CDF was usually caused by cholelithiasis (573 of 728). Epigastric pain (589 of 728) and cholangitis (395 of 728) were the most common symptoms of CDF. CDF was usually detected and confirmed by endoscopic retrograde cholangiopancreatography (ERCP) (475 of 728) in Mainland China. The fistulas larger than 1 cm (82 of 654) were recommended for surgical biliary reconstruction. Fistulas between 0.5 cm and 1.0 cm (467 of 654) which were followed frequently by cholangitis attacks also required surgery; the rest were recommended to have stone removal and/or the application of an effective biliary drainage. Fistulas less than 0.5 cm (105 of 654) were usually received conservative therapy. CONCLUSION: CDF should be considered in differential diagnosis of recurrent epigastric pain and cholangitis. A possible ERCP should be arranged to investigate carefully. Depending on the size of fistula and clinical presentation, different programs for CDF are indicated, ranging from drug therapy to choledochojejunostomy.

Therapeutic effect and safety of ustekinumab for plaque psoriasis: a meta-analysis / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of ustekinumab in the therapy of plaque psoriasis.</p><p><b>METHODS</b>Literatures published up to November 2013 were collected from Cochrane library, MEDLINE, and PubMed which were related with ustekinumab for plaque psoriasis. The efficacy was estimated using relative risk of Psoriasis Area and Severity Index (PASI) 75 response rate at the week 12 endpoint in clinical trials, and adverse effects were also analyzed. Meta-analysis was carried out by using Review Manager 5.1.</p><p><b>RESULTS</b>Six randomized control trials consistent with the inclusion criteria were selected and reviewed. Ustekinumab 45 mg group and 90 mg group could get better therapeutic effect compared with the placebo group (all P<0.00001). Furthermore, ustekinumab 90 mg group was more effective than ustekinumab 45 mg group (P=0.01). Adverse effects in the 6 trials were mentioned including headache, upper respiratory tract infection, nasopharyngtis, infection, serious infection, cardiovascular events, and malignant tumors. There were no statistically significant differences of these adverse effects among three groups (all P>0.05), except that infection rate in ustekinumab 45 mg group was higher than the placebo group (P=0.02).</p><p><b>CONCLUSIONS</b>Ustekinumab is an effective and safe therapeutic method for plaque psoriasis. However, further longer time analysis of safety is needed.</p>

The fifth metastasis route of gastric cancer and the third principle of radical operation / 中华胃肠外科杂志

The local-regional recurrence is the most important failure reason of D2 radical operation for curative advanced gastric cancer. Another way of metastasis, which exists in the mesogastrium and separates apart from the primary tumor, blood vessels and lymph nodes, appears to be responsible for the local-regional recurrence partly. We call it as the fifth route of metastasis. The mesogastrium is surrounded by proper fascia completely and covered partly by serosa. We suggest that D2 radical operation should not only include the resection of primary tumor and lymphadenectomy, but also add the complete mesogastric excision (CME) as the third principle in gastric cancer radical operations to cut down the fifth metastasis route of gastric cancer, to prevent the fifth metastasis scattered on the field of operation which may induce the local-regional recurrence.

Types of primary cyclins expressed in gastric carcinoma and their relationship with clinicopathological features / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To characterize the human primary cyclins (D1, E, A, B1) expressed in gastric carcinoma, and to clarify the relationship between the types of expressed primary cyclins and clinicopathological features of gastric carcinoma.</p><p><b>METHODS</b>Primary cyclins (D1, E, A, B1) expressed in single cells separated from 68 cases gastric carcinoma tissues were analyzed by flow cytometry. We classified the gastric carcinomas by different types of the expressed primary cyclins, and explore the roles of primary cyclins expressed in cell cycle and the expression patterns of the cyclins. The results were analyzed together with clinicopathological features.</p><p><b>RESULTS</b>The patterns of expressed primary cyclins could be classified into five types. The proportion was 10.3% (7/68), 22.1% (15/68), 25.0% (17/68), 29.4% (20/68), and 13.2% (9/68), respectively, from type I to type V. Each type could be, according to the degree of in-cycle cyclins expressed, divided into different sub-types. The types of primary cyclins expressed were strongly linked to invasive depth and lymph node metastasis of the gastric carcinoma (P < 0.01). The rates of lymph node metastasis were 26.6%, 43.8%, 82.3%, 95.0%, and 100.0%, respectively, from type I to type V. The type of primary cyclins expressed was also significantly associated with disease stage (TNM stage). The proportion of stage IV disease was 0, 6.7%, 17.6%, 25.0% and 55.6%, respectively, from type I to type V. It was shown that there were relationships between the sub-types of primary cyclins expressed and different growth-types, degree of cell differentiation, or, the tumor gross types (P < 0.01).</p><p><b>CONCLUSIONS</b>The types of primary cyclins expression are different in the process of the occurrence, development and metastasis of gastric carcinoma, and are correlated with clinicopathological features of gastric carcinoma.</p>

Remarkably reduced expression of FoxO3a in metaplastic colorectum, primary colorectal cancer and liver metastasis / 华中科技大学学报（医学）（英德文版）

The forkhead family members of transcription factors (FoxOs) are expected to be potential cancer-related drug targets and thus are being extremely studied recently. In the present study, FoxO3a, one major member of this family, was identified to be down-regulated in colorectal cancer through micro-array analysis, which was confirmed by RT-PCR and Western blot in 28 patients. Moreover, immunohistochemistry (IHC) showed that the expression levels of FoxO3a were remarkably reduced in 99 cases of primary colorectal cancer, liver metastasis, and even in metaplastic colorectal tissue. IHC also revealed an exclusion of FoxO3a from the nucleus of most cells of tumor-associated tissues. Silencing FoxO3a by siRNA led to elevation of G2-M phase cells. We conclude that the downregulation of FoxO3a may greatly contribute to tumor development, and thus FoxO3a may represent a novel therapeutic target in colorectal cancer.

Remarkably reduced expression of FoxO3a in metaplastic colorectum, primary colorectal cancer and liver metastasis / 华中科技大学学报（医学）（英德文版）

The forkhead family members of transcription factors (FoxOs) are expected to be potential cancer-related drug targets and thus are being extremely studied recently. In the present study, FoxO3a, one major member of this family, was identified to be down-regulated in colorectal cancer through micro-array analysis, which was confirmed by RT-PCR and Western blot in 28 patients. Moreover, immunohistochemistry (IHC) showed that the expression levels of FoxO3a were remarkably reduced in 99 cases of primary colorectal cancer, liver metastasis, and even in metaplastic colorectal tissue. IHC also revealed an exclusion of FoxO3a from the nucleus of most cells of tumor-associated tissues. Silencing FoxO3a by siRNA led to elevation of G2-M phase cells. We conclude that the downregulation of FoxO3a may greatly contribute to tumor development, and thus FoxO3a may represent a novel therapeutic target in colorectal cancer.

Indication of gastrointestinal laparoscopy / 中华胃肠外科杂志

As the evidence-based medicine of gastrointestinal laparoscopy (GIL) for gastrointestinal cancer accumulated, the procedures have become a "should be done" treatment, not a "can be done" ones, for early or median gastric or colorectal cancer. So, we propose that GIL for cancer should be indicationization, rather than doctorization. The former is that the procedures should be performed according to indication of the gastrointestinal tumors, and the latter is that the different procedures applied depend on the doctor's experience. For indicationization of GIL, special training program is suggested and format or standardization of GIL for cancer should be abstracted.

Expressions and significance of COX-2 and P-gp in human hepatocellular carcinoma tissues / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effect of the COX-2 gene on the oncogenesis and development of the hepatocellular carcinoma and the influence of COX-2 gene on the expression of P-gp protein.</p><p><b>METHOD</b>Fifty-two pieces of the hepatocellular carcinoma samples and 20 cases of normal liver samples were collected from the patients operated from October 2003 to June 2005. RT-PCR and immunohistochemistry staining were employed to detect the COX-2 mRNA as well as P-gp protein in the normal liver tissues and the carcinoma tissues. Meanwhile, the expression of the mdr 1 mRNA in the carcinoma tissues was also determined and the correlation between the expressions of the COX-2 and P-gp was investigated.</p><p><b>RESULTS</b>No expression of the COX-2 in the normal liver tissue was detected. The positive expression of COX-2 in the low and middle differentiated carcinoma was elevated significantly as compared with that in the high differentiated carcinoma tissue (x2 = 6.80, P less than 0.01). The positive expression of the COX-2 in the HBSAg (+) carcinoma tissue was significantly higher as compared with that in the HBSAg (-) carcinoma (x2 = 4.70, P less than 0.05), and the carcinoma in combination with cirrhosis also showed significantly higher in expression of COX-2 than the carcinoma without cirrhosis (x2 = 7.51, P less than 0.01). The mdr1 mRNA was found both expressed in the normal and carcinoma tissues. The expression of COX-2 mRNA was found in the carcinoma, but not found in the normal tissues. The COX-2 mRNA and mdr1 mRNA was found both expressed in the normal and carcinoma tissues. The correlation coefficient between COX-2 and mdr1 mRNA was 0.563 ( P less than 0.01).</p><p><b>CONCLUSION</b>The results indicated that Cox-2 gene might involved in the multidrug resistance of the hepatocellular carcinoma mediated by P-gp.</p>

Relationship of NOR-1 with the regulation of inflammation via liver X receptor alpha in Kupffer cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the relationship of NOR-1 with the inhibition of inflammatory reaction in mice Kupffer cells (KCs) induced by lipopolysaccharide (LPS) via liver X receptor alpha (LXR alpha).</p><p><b>METHODS</b>KCs from male KM mice were isolated by density gradient centrifugation, incubated and then randomly assigned to three groups: control group, LPS treated group and LPS+T0901317 treated group.</p><p><b>RESULTS</b>The mRNA and protein expressions of LXR alpha and NOR-1 in each group were determined by RT-PCR, immunofluorescent assay and western blot, respectively. The densities of TNF alpha and IL-10 in supernatants were evaluated by enzyme linked immunosorbent assay (ELISA). The mRNA and protein expression levels of LXR alpha in LPS + T0901317 group were the highest as compared to the other two groups (0.748+/-0.072 and 1.217+/-0.133 respectively), The mRNA and protein expression levels of NOR-1 in LPS+ T0901317 group were the highest as compared to the other two groups (2.726+/-0.065 and 0.842+/-0.058 respectively). The densities of supernatant TNF alpha in LPS group and IL-10 in LPS+T0901317 group were the highest [(450.89+/-78.52) ng/L and (537.41+/-36.41) ng/L respectively].</p><p><b>CONCLUSIONS</b>Promoting the expression of LXR alpha in KCs can elevate the NOR-1 expression and then inhibit inflammatory reaction.</p>

Tacrolimus alleviates acute liver graft rejection by inhibiting glucocorticoid-induced tumor necrosis factor-related protein ligand in rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the mechanism underlying the inhibitory effect of tacrolimus (FK506) against acute liver graft rejection.</p><p><b>METHODS</b>Rat models of orthotopic liver transplantation were divided into 3 groups, namely the tolerance group with Brown Norway (BN) rats as the donors and Lewis rats as the recipients, rejection group with Lewis rats as donors and BN rats as recipients, and FK506 group with the same donor-recipient pair as in the rejection group and FK506 treatment. The recipients were sacrificed 7 days after the transplantation, and the hepatic histology, cytokine levels, and glucocorticoid-induced tumor necrosis factor-related protein ligand (GITRL) expression in the liver and Kupffer cells were observed and detected.</p><p><b>RESULTS</b>Compared with the tolerance group, the rejection group showed increased GITRL expressions in the liver and Kupffer cells (P<0.05), which was significantly lowered by FK506 treatment (P<0.05). Acute liver graft rejection caused significantly elevated interferon-γ (IFN-γ) levels and decreased interleukin-10 (IL-10) levels in the plasma and Kupffer cells (P<0.05), and these changes were obviously attenuated by FK506 treatment (P<0.05).</p><p><b>CONCLUSION</b>The effect of FK506 in suppressing acute liver graft rejection is probably associated with down-regulated GITRL expression in the liver and Kupffer cells.</p>

Recombinant adenovirus with human indoleamine-2,3-dioxygenase and hepatitis B virus preS was constructed and expressed in HepG2 cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Indoleamine-2,3-dioxygenase (IDO) is proven to suppress hepatitis B virus (HBV) specific immune response and depletion of IDO may be a useful approach for HBV therapy. To test this concept, we constructed recombinant adenovirus with human IDO and HBV preS, which would form the basis for future in vivo experiments.</p><p><b>METHODS</b>The fragment of human IDO and HBV preS cDNA were subcloned into multiple cloning sites in an adenoviral vector system containing two cytomegalovirus (CMV) promoters. Recombination was conducted in the Escherichia coli BJ5183. The recombinant adenovirus containing hIDO gene and HBVpreS gene was packaged and amplified in 293 cells. Integration was confirmed by polymerase chain reaction as well as the quantification of viral titers. HepG2 cells were infected with the recombinant adenovirus and mRNA and protein specific for hIDO and HBVpreS was detected by RT-PCR and Western blotting respectively.</p><p><b>RESULTS</b>The recombinant adenovirus was produced successfully. Its titer was 2.5 × 10(9) efu/ml. IDO and HBVpreS mRNA as well as the encoded proteins could be found in transfected HepG2 cells, but not in control HepG2 cells.</p><p><b>CONCLUSION</b>The transfer of hIDO-HBVpreS with double-promoter adenoviral vector was efficient. The recombinant adenovirus with hIDO and HBV preS would provide the experimental basis for future studies.</p>