Jujuboside B alleviates acetaminophen-induced hepatotoxicity in mice by regulating Nrf2-STING signaling pathway.

BACKGROUND: Jujuboside B (JuB) is the main bioactive saponin component of Chinese anti-insomnia herbal medicine Ziziphi Spinosae Semen, which has been reported to possess varied pharmacological functions. Even though it has been traditionally used to treat inflammation- and toxicity-related diseases, the effects of JuB on acetaminophen (APAP) overdose-induced hepatotoxicity have not been determined yet. METHODS: C57BL/6 J mice were pre-treated with JuB (20 or 40 mg/kg) for seven days before APAP (400 mg/kg) injection. After 24 h of APAP treatment, serum, and liver tissues were collected to evaluate the therapeutic effects. To investigate whether the Nrf2-STING signaling pathway is involved in the protective effects of JuB against APAP-induced hepatotoxicity, the mice received the DMXAA (the specific STING agonist) or ML385 (the specific Nrf2 inhibitor) during the administration of JuB, and Hematoxylin-eosin staining, Real-time PCR, immunohistochemical, and western blot were performed. RESULTS: JuB pretreatment reversed APAP-induced CYP2E1 accumulations and alleviated APAP-induced acute liver injury. Furthermore, JuB treatment significantly inhibited oxidative stress and the pro-inflammatory cytokines, as well as alleviated hepatocyte apoptosis induced by APAP. Besides, our result also demonstrated that JuB treatment upregulated the levels of total Nrf2, facilitated its nuclear translocation, upregulated the expression of HO-1 and NQO-1, and inhibited the APAP-induced STING pathway activation. Finally, we verified that the beneficial effects of JuB were weakened by DMXAA and ML385. CONCLUSION: Our study suggested that JuB could ameliorate APAP-induced hepatic damage and verified a previously unrecognized mechanism by which JuB prevented APAP-induced hepatotoxicity through adjusting the Nrf2-STING pathway.

PD-1/PD-L1 inhibition promotes hepatic regeneration in small-for-size liver following extended hepatectomy.

BACKGROUND: Small-for-size syndrome following liver surgery is characterized by compromised liver regeneration. Liver macrophages play key roles in initiating liver regeneration, and modulation of the immune microenvironment through macrophages may accelerate liver regeneration. In our current study, we aimed to explore the involvement of innate immunity after extended hepatectomy in rats and humans, and to test the effect of immunity modulation on small-for-size liver regeneration in rats. METHODS: Serum programmed cell death protein ligand 1 (PD-L1) was measured after major hepatectomy and minor hepatectomy in humans and rats. Liver regeneration in rats was assessed using liver-to-body weight ratio and kinetic growth rate, antigen Ki67 and proliferating cell nuclear antigen (PCNA), and macrophage polarization was assessed by inducible nitric oxide synthase (iNOS), cluster of differentiation protein 163 (CD163) expression by immunohistochemistry (IHC) and iNOS/CD163 ratio. Rat hepatocyte BRL or human hepatocyte LO2 were co-cultured with rat bone marrow-derived macrophages or human macrophages THP-1. BMS-1 or Nivolumab were used to block programmed cell death protein 1 (PD-1)/PD-L1 in vitro and in vivo. RESULTS: PD-L1 expressions were significantly higher following major hepatectomy compared to minor resection in both humans and rats; compromised liver regeneration after extended hepatectomy in rats was associated with PD-L1 upregulation and M2 macrophage polarization. M1 macrophages increased proliferation of hepatocytes through interleukin-6 (IL-6), and M2 macrophages decreased hepatocyte proliferation; blocking PD-1/PD-L1 reversed the effect of M2 macrophages on the survival of hepatocytes in vitro and promoted liver growth in rats through M1 macrophage polarization. CONCLUSION: Compromised hepatic regeneration following extended hepatectomy is characterized by M2 macrophage polarization and upregulated PD-L1 expression. Blocking PD-1/PD-L1 may enhance small-for-size liver regeneration by inducing M1 macrophage polarization.

[Status quo of autologous sperm preservation in the human sperm bank in Beijing].

OBJECTIVE: To comprehensively analyze the status quo of autologous sperm preservation in the human sperm bank in Beijing and better utilize the existing resources for the preservation of male fertility. METHODS: We retrospectively analyzed the geographical data and semen quality of 251 males with autologous sperm preservation in the Human Sperm Bank, Science and Technology Research Institute, National Health and Family Planning Commission of China from July 2006 to December 2016. RESULTS: The rate of autologous sperm preservation in the Human Sperm Bank was as low as 8.76% between July 2006 and December 2010 but increased annually by 119% on average from 2011 to 2013. Of the 251 males involved, 204 (81.27%) were aged 20－39 years, 175 (69.72%) had bachelor's or master's degree, 223 (88.84%) had no child, 69 (27.49%) got less than 10 tubes of semen samples frozen, and 26 (10.36%) had their semen samples cryopreserved only once. The utilization rate of the cryopreserved sperm was only 5.58 % (n = 14). The main reason for autologous sperm preservation was carcinoma (55.78% ï¼»n = 140ï¼½), including blood cancer (22.31% ï¼»n = 66ï¼½), testicular cancer (13.15% ï¼»n = 33ï¼½) and other cancers (16.33% ï¼»n = 41ï¼½). Compared with the non-cancer males, the cancer patients had a significantly reduced mean sperm concentration (90.45 vs 60.53 ×106/ml, P < 0.05), total sperm count (311.3 vs 175.8 ×106, P < 0.05), percentage of progressively motile sperm (PMS) (49.21% vs 43.55%, P < 0.05) and recovery rate of PMS (68.13% vs 52.17%, P < 0.05). In the subgroups of testicular, blood and other cancers, the sperm concentration averaged 37.68, 57.98 and 90.69 ×106/ml, the semen volume 2.73, 2.82 and 3.41 ml, the total sperm count 93.29, 158.41 and 349.49 ×106, the percentage of PMS 45.32%, 43.47% and 44.49%, and the recovery rate of PMS 48.32%, 50.07% and 61.09%, respectively, the sperm concentration and total sperm count significantly lower in the testicular cancer patients than in the other two groups (P < 0.05). CONCLUSIONS: The number of the cases of autologous sperm preservation in Beijing is increasing year by year, and the majority of them are cancer patients. As most of the cancer patients have missed the best period for sperm preservation, sperm bank workers should endeavor to increase the public awareness of autologous sperm preservation.

Effects of C-myc gene silencing on interleukin-1ß-induced rat chondrocyte cell proliferation, apoptosis and cytokine expression.

This study explores the effects of C-myc gene silencing on cell proliferation, apoptosis and cytokine expression in interleukin (IL)-1ß-induced rat chondrocytes. Primary chondrocytes were obtained from 40 Sprague-Dawley rats. For in vitro C-myc3-shRNA transfection, chondrocytes were assigned to a blank 1, model 1, IL-1ß + C-myc3-shRNA, C-myc3-shRNA, (IL-1ß + C-myc3-shRNA) + C-myc overexpression, C-myc3-shRNA + C-myc overexpression or IL-1ß + C-myc-Con group. Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR) were performed to detect C-myc, PCNA and cyclin D1 mRNA and protein expression. Cell proliferation was analyzed via CCK-8 assay and cell cycle while apoptosis was measured through flow cytometry. ELISA was utilized to assess the levels of metallopeptidase 13 (MMP-13), IL-6 and tumor necrosis factor-α (TNF-α). Both the qRT-PCR and Western blotting results demonstrated that C-myc3-shRNA transfection inhibits C-myc expression and promotes PCNA and cyclin D1 expression. In comparison to the model 1 group, all groups except the (IL-1ß + C-myc3-shRNA) + C-myc overexpression and IL-1ß + C-myc-Con groups showed increases in cell proliferation and S phase cell count and decreases in G0/G1 phase cell count, cell apoptosis and MMP-13, IL-6 and TNF-α levels. The model 1, C-myc3-shRNA and C-myc3-shRNA + C-myc overexpression groups displayed higher cell proliferation and S phase cell count and reduced G0/G1 phase cell count, cell apoptosis and MMP-13, IL-6 and TNF-α levels than the IL-1ß + C-myc3-shRNA group. In comparison to the model 1 and C-myc3-shRNA + C-myc overexpression groups, the C-myc3-shRNA group promoted cell proliferation and S phase cell counts but suppressed G0/G1 phase cell count, cell apoptosis and MMP-13, IL-6 and TNF-α levels. In conclusion, the study demonstrates that C-myc gene silencing can promote cell proliferation and inhibit cell apoptosis and cytokine expression in IL-1ß-induced rat chondrocytes.

Urban ecological security pattern construction coupled with risk and service: A case study of Xianyang City, Shaanxi Province, China.

How to combine regional ecological risks and local ecological needs to construct ecological security is one of the main issues of its application in territorial spatial governance and associated with whether it can be effectively applied. Based on the "source" accessibility and the quality of space, we constructed the ecological security pattern of Xianyang City through the way of source-corridor-node. During the construction processes, we combined the cha-racteristics of topography, influencing factors of regional ecological security, and landscape characteristics. We coupled them with morphological spatial pattern analysis (MSPA) and comprehensive evaluation results of ecological resistance, ecological connectivity and ecosystem service value, and superimposed with ecological gradient ana-lysis. The results showed that there were 66 ecological sources, with a total area of 2506.65 km2, accounting for 24.6% of the total area of Xianyang City, which were mainly distributed in the northeast, west and central mountainous areas. There were 106 ecological corridors with a total length of 823.5 km, including potential corridors, water systems, irrigation canal sites, Qinzhidao and other natural and cultural systems, which extended along the ecological source to the northwest and south Weihe River. There were 20 ecological nodes to improve ecological connectivity, which were mainly distributed between second layers of loess tableland and arid mountainous areas with banded distribution in the north part of the city.

Retrospective comparison of titanium elastic nail (TEN) and reconstruction plate repair of displaced midshaft clavicular fractures.

BACKGROUND: Titanium elastic nails (TENs) are commonly used to repair displaced midshaft clavicular fractures. Although several clinical studies have shown that the use of TENs is more effective and less invasive than traditional plate fixation, high rates of complications associated with TENs have been reported. This retrospective study compared the use of TENs with reconstruction plates in the treatment of displaced midshaft clavicular fractures. MATERIALS AND METHODS: From January 2005 to July 2007, 141 patients with displaced midshaft clavicular fractures were treated with TEN or plate fixation. At 6 and 24 months postoperatively, we evaluated the patients' general health, operative complications, and functional recovery. RESULTS: The mean bone union time of TEN patients was 12.4 ± 3.4 weeks, whereas that in the plate group was 14.4 ± 3.7 weeks. The time of union was significantly shorter in the TEN group than that in the plate group. There was no significant difference between these 2 groups' rates of complications such as nonunion or malunion. Shoulder function scores were significantly better in the TEN group than in the plate group at 6 months postoperatively. There was no significant difference at 24 months postoperatively. Patients were more satisfied with the cosmetic appearance and overall outcome after TEN repair than after plate fixation. CONCLUSIONS: TEN fixation of displaced midshaft clavicular fractures allows for a faster functional recovery, higher patient satisfaction, and a more cosmetically satisfactory appearance than plate fixation. The complication rates of both TENs and reconstruction plates were similar.

[Arthroscopic treatment of ankle instability combined with anteromedial impingement syndrome].

OBJECTIVE: To investigate the surgical skills and clinical curative results of arthroscopic treatment of ankle instability combined with anteromedial impingement syndrome. METHODS: From February 2019 to August 2020, 13 patients with ankle instability combined with anteromedial impingement were retrospectively analyzed. There were 10 males and 3 females with age of (40.0±15.1) years old. The course of disease was(44.1±33.2) months. All patients had history of ankle sprain. MRI showed the injury of anterior talofibular ligament. All patients had anteromedial pain and pressing pain when ankle dorsiflexion. All patients were treated with ankle debridement and Brostr?m-Gould surgery under ankle arthroscopic. Postoperative results were evaluated by VAS(visual analogue scale) and AOFAS-AH(American Orthopaedic Foot and Ankle Society Ankle-Hindfoot scale, AOFAS-AH). RESULTS: All 13 patients completed the surgery successfully with an operative time of 60 to 90 minutes. All the surgical incisions healed by first intention, and no complications such as incision infection, skin necrosis and neurovascular injury. Follow-up time was (18.1±4.7) months. At the latest follow-up, the VAS score was 1.2±1.1, which was significantly lower than the preoperative score 4.8±1.5 (P<0.05);the AOFAS-AH score 94.2±5.1 was significantly higher than the preoperative score 65.5±11.5 (P<0.05). The AOFAS-AH score at the final follow-up ranged from 84 to 100. All patients walked with normal gait without ankle instability or impingement recurrence. CONCLUSION: Ankle anteromedial impingement syndrome combined with ankle instability is easy to be ignored clinically. Such kind of anteromedial impingement syndrome is mostly related to osteophyte at dorsal medial talar neck. Arthroscopic treatment of ankle instability combined with anteromedial impingement syndrome has satisfactory curative effect with safety and minimal injury.

[Expression of liver-intestine cadherin and its significance in hepatocellular carcinoma].

OBJECTIVE: To explore the expression and clinical significance of LI (liver-intestine)-cadherin mRNA and protein expression level in hepatocellular carcinoma. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical techniques were used to detect the expression of the LI-cadherin mRNA and protein in 56 cases of hepatocellular carcinoma, 44 cases of corresponding paracancerous tissues and 9 cases of normal liver tissues. The relationships with its clinicopathological characteristics were analyzed. RESULTS: The mRNA of LI-cadherin was expressed in 56 cases of hepatocellular carcinoma tissues and 44 cases of corresponding paracancerous tissues. But its expression was not detected in normal tissues. Semiquantitative analysis showed that the mRNA expression level of LI-caherin was greater in hepatocellular carcinoma tissues than that in corresponding paracancerous tissues (0.653 ± 0.147 vs 0.534 ± 0.138). The differences were statistically significant (P < 0.01). The positive rates of LI-cadherin protein were 64.29% (36/56) in hepatocellular carcinoma tissues, 22.73% (10/44) in paracancerous tissues and 0% (0/9) in normal tissues. CONCLUSION: The expression of LI-cadherin is up-regulated significantly in HCC. The over-expression of LI-cadherin protein is correlated with the lymph node metastasis and tumor thrombi in portal vein. It indicates that LI-cadherin may play an important role in the progression and metastasis of HCC.

Clinical outcomes of midclavicular fractures treated with titanium elastic nails.

BACKGROUND: Midclavicular fractures are common clinically, accounting for about 76% of all clavicular fractures. Recent studies have revealed a previously unrecognized incidence of nonunion and malunion after conservative treatment of more severe midclavicular fractures. Our aim was to evaluate the clinical outcomes of midclavicular fractures treated with titanium elastic nails. METHODS: From February 2005 to February 2007, 41 patients with displaced midclavicular fractures received open reduction and internal fixation with a titanium elastic nail inserted through the sternal end of the clavicle. We evaluated the visual analogue scale (VAS), the Constant score and the Disabilities of the Arm, Shoulder and Hand (DASH) scale to determine outcomes. RESULTS: A mean follow-up of 14.5 months (range 7-24 mo) revealed radiographic fracture union in all patients with an average clinical healing time of 2.2 months. Mean subjective pain 3 days after surgery was significantly lower than the day before surgery (p < 0.001). The mean range of motion 3 days after surgery was significantly improved compared with the day before surgery (p < 0.001). The nails were removed in all patients a mean of 7.2 months (range 5.4-9.5 mo) after surgery, and no fractures recurred. The mean postoperative DASH score was 2.5 (range 0.5-8.0) and the mean postoperative Constant score was 95.2 (range 86.5-97.0). CONCLUSION: Limited open reduction and internal fixation with titanium elastic nails is a safe and minimally invasive surgical procedure for the treatment of displaced midclavicular fractures in adults and achieves good functional results and high patient satisfaction.

Characterization and expression of a new subfamily member of penaeidin antimicrobial peptides (penaeidin 5) from Fenneropenaeus chinensis.

Penaeidins are members of a special family of antimicrobial peptides existing in penaeid shrimp and play an important role in the immunological defence of shrimp. Here, we report one penaeidin with a putative isotype newly cloned from fleshy prawn Fenneropenaeus chinensis. The penaeidin open reading frame encodes a 79 amino acid peptide while two exons and an intron were identified within the 1126bp genomic sequence of Fenchi-penaeidin 5. Phylogenetic analysis and sequence comparison with other known penaeidins suggest the new gene belongs to a novel subfamily of penaeidins and the two isoforms were named Fenchi-penaeidin 5-1 and 5-2, respectively. Fenchi-penaeidin 5 mRNA was examined in normal and microbial challenged shrimp and was found to be constitutively expressed in heamocytes, heart, gill, intestine and ovary. Bacterial challenge resulted in mRNA up-regulation, inducing expression in hepatopancreas and stomach. Fenchi-penaeidin 5-1 was also expressed in Pichia pastoris, and recombinant Fenchi-penaeidin 5-1 exhibited activities against Gram-positive and -negative bacteria and fungi.

Hyaluronic acid, an efficient biomacromolecule for treatment of inflammatory skin and joint diseases: A review of recent developments and critical appraisal of preclinical and clinical investigations.

Hyaluronic acid (HA) plays multifaceted role in regulating various biological processes and maintaining homeostasis into the body. Numerous researches evidenced the biomedical implications of HA in skin repairmen, cancer prognosis, wound healing, tissue regeneration, anti-inflammatory, immunomodulation. The present review was aimed to summarize and critically appraise the recent developments and efficacy of HA for treatment of inflammatory skin and joint diseases. A thorough analysis of the literature revealed that HA based formulations (i.e., gels, creams, autologous graft, thin sheets, soaked gauze, gauze pad, tincture, injection) have shown remarkable efficacy in treating a wide range of inflammatory skin diseases. The safety, tolerability, and efficacy of HA (as intra-articular injection) have also been well-documented for treatment of various types of joint disease including knee osteoarthritic, joint osteoarthritis, canine osteoarthritis, and meniscal swelling. Intra-articular injection of HA produces remarkable reduction in joint pain, synovial inflammation, and articular swelling. A remarkable improvement in chondrocyte density, territorial matrix appearance, reconstitution of superficial amorphous layer of the cartilage, collagen remodelling, and regeneration of meniscus have also been evident in patients treated with HA. Conclusively, we validate that the application/administration of HA is a promising pharmacotherapeutic regimen for treatment of inflammatory skin and joint diseases.

Inhibition of PI3K/AKT/mTOR signaling pathway promotes autophagy of articular chondrocytes and attenuates inflammatory response in rats with osteoarthritis.

OBJECTIVE: This study aims to explore the relationship between PI3K/AKT/mTOR signaling pathway and autophagy of articular chondrocytes in rats with osteoarthritis (OA). METHODS: Rat articular chondrocytes were isolated and cultured, and then induced by protein inhibitors of PI3K/AKT/mTOR signaling pathway. Chondrocytes were assigned into blank group, IL-1ß induction group (IL-1ß group), PI3K inhibitor+IL-1ß induction group (PI3Ki+IL-1ß group), AKT inhibitor+IL-1ß induction group (AKTi+IL-1ß group) and mTOR inhibitor+IL-1ß induction group (mTORi+IL-1ß group). Cell proliferation activity was detected by MTT assay, cell cycle by flow cytometry and cell autophagy by monodansylcadaverine (MDC) staining. Autophagy rates were evaluated by GFP-LC3 fluorescence microscopy. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to detect mRNA expressions of autophagy-related genes (Atg5 and Atg7). Western blotting was utilized to detect expressions of autophagy markers (LC3, Beclin1 and p62) and of relevant proteins in the PI3K/AKT/mTOR signaling pathway. RESULTS: The cell proliferation rate of the IL-1ß group was lower than that of the blank group after cells were cultured for 24h, and the cell proliferation rates of the PI3Ki+IL-1ß group, the AKTi+IL-1ß group and the mTORi+IL-1ß group were higher than those of the IL-1ß group. In comparison with the blank group, cells in the IL-1ß group were arrested at the G1 phase and decreased in the S phase, MDC positive staining cells were decreased with attenuated staining intensity, the autophagy rate was decreased, the mRNA expressions of Atg5 and Atg7 and the protein expressions of LC3, Beclin1 and p62 were significantly down-regulated. While in the groups of PI3Ki+IL-1ß, AKTi+IL-1ß and mTORi+IL-1ß, haploid cells were reduced, coupled with an increased proportion of cells in the S phase and decreased proportion of cells in the G1 phase, the autophagy rate was increased, the mRNA expressions of Atg5 and Atg7 and the protein expressions of LC3, Beclin1 and p62 were significantly up-regulated. Compared with the blank group, the protein phosphorylation levels of PI3K, AKT and mTOR were elevated, while there were no significant difference observed in the total amount of PI3K, AKT and mTOR in the IL-1ß group. Meanwhile, there were relatively low protein phosphorylation levels of PI3K, AKT and mTOR in the groups of PI3Ki+IL-1ß, AKTi+IL-1ß and mTORi+IL-1ß. CONCLUSIONS: Inflammation could inhibit the proliferation and cell cycle of rat chondrocytes and reduce the autophagy rate. Inhibition of PI3K/AKT/mTOR signaling pathway could promote the autophagy of articular chondrocytes and attenuate inflammation response in rats with OA.

Molecular cloning and characterization of cecropin from the housefly (Musca domestica), and its expression in Escherichia coli.

A cDNA encoding housefly (Musca domestica) cecropin transcript was isolated from total RNA using RT-PCR, 3'RACE, and lambda gt11 cDNA library screening, and was expressed in Escherichia coli. This is the first report of a cecropin nucleotide and amino acid sequence in the housefly. The open reading frame of Md-Cec (189bp) encodes a precursor of 63 aa, which is comprised of a 23 aa signal peptide and a 40 aa mature peptide. In terms of amino acid sequence, Md-Cec shares a high degree of identity (74-82%) with those of some Diptera insects. Northern blot, RT-PCR and in situ hybridization analyses revealed that the prececropin was temporally expressed 5h after bacteria-challenge in larvae, and was induced in the fat body, epithelia of the body wall, and the epidermis of the midgut. The DNA fragment encoding mature Md-Cec was then subcloned into the pGEX-4T-1 expression vector and was highly expressed in E. coli BL21 with IPTG induction. The expressed proteins, fused to glutathion S-transferase, were purified by glutathion-Sepharose 4B affinity chromatography and cleaved with thrombin, followed by gel filtration chromatography. Recombinant Md-Cec exhibited antimicrobial activity against E. coli.

Surgical treatment of hepatocellular carcinoma originating from caudate lobe--a report of 39 cases.

The study objective was to study the therapeutic effect of surgical treatment for hepatocellular carcinoma (HCC) originating from the caudate lobe. From 1995 to 2003, caudate lobe resection was carried out for 97 cases; among them 39 were for HCC, who were divided into two groups. Group A consisted of 19 cases undergoing isolated caudatectomy, and group B consisted of 20 cases undergoing caudatectomy combined with other liver resections. The factors that might influence postoperative recovery were compared between the two groups. A special instrument, Peng's Multifunctional Operative Dissector, was used for surgical dissection. All tumors were resected successfully. One patient died of postoperative renal failure. Hydrothorax occurred in three patients, ascites occurred in four patients, and bile leakage occurred in one patient. Thirty cases received long-term follow-up with survival rates at 1, 3, and 5 years of 53%, 50%, and 39%, respectively. Caudate lobectomy is an effective therapeutic method for HCC originating in the caudate lobe. Isolated caudatectomy should be performed as the first choice whenever possible. Anterior transhepatic approach is appropriate in some cases. Peng's Multifunctional Operative Dissector is a very useful instrument for surgical dissection.

[Establishment of liver metastasis model of human gallbladder cancer and isolation of the subpopulation with high metastatic potential].

OBJECTIVE: To establish an experimental liver metastatic model of gallbladder cancer and isolate the subpopulation with high metastatic potential from the model, which may serve as a reliable tool in research on liver metastasis of gallbladder cancer in vivo and in vitro. METHODS: Human gallbladder cancer cells of the line GBC-SD were cultured. Ten nude athymic BALB/c mice, aged 4 approximately 5 weeks, underwent inoculation of suspension of GBC-SD cells into the spleens and then their spleens were resected. Three weeks later laparotomy was performed to observe if liver metastasis occurred. Once liver metastasis was discovered, the mice were killed and the livers were taken out to undergo microscopy, tumor cells were isolated from the metastatic foci and cultured in vitro, and then inoculated into other 10 mice in the same way as their parents cells for the second round of selection. The similar steps were repeated, altogether for three rounds of selection and a total 30 mice were inoculated in 3 rounds so as to find subpopulation with high metastatic potential. Another 10 mice underwent subcutaneous inoculation, 90 days later the mice were killed to observe if pulmonary metastasis occurred. PCR was used to detect the 3 microsatellite sequences D14S68, D18S69, and D20S199 in the DNA samples of the gallbladder cancer cells of the parent cell line GBC-SD, the isolated cells of the subpopulation with high metastatic potential, and the liver of experimental animal. RESULTS: 90% of the mice inoculated subcutaneously with the GBC-SD cells developed subcutaneous tumors, however, no mouse in this group died and no pulmonary metastasis was found. The liver metastatic rate was 65% in the 10 mice undergoing intrasplenic inoculation. Thus a metastatic model of human gallbladder cancer in nude mice was established. The liver metastatic tumors were uniformly distributed throughout the liver parenchyma with predominance to the periphery. Despite multiple sites of involvement, the left lobes were most commonly affected in all experimental animals. Histological examination of the metastatic lesion demonstrated adenocarcinoma. Gross hepatic metastasis was detected 10, 7, and 5 weeks after the inoculation respectively in the first, second, and third round selection respectively with an incidence rate of metastases of 60%, 70%, and 90% respectively. From the third round metastatic model a subpopulation with high metastatic potential was isolated and designated as GBC-SD/M, which exhibited the similar histological characteristics as its parent cell line GBC-SD under inverted light microscopy. The three amplified bands at the sites 14S68, D18S69, and D20S199, amplified with the three pairs of specific primers for the three homo-specific microsatellites, were detected in the GBC-SD cells and GBC-SD/M cells, but not in the liver of tumor-bearing animal. CONCLUSION: A liver metastasis model of human gallbladder cancer has been established, a reliable and efficient tool for study on the metastasis of gallbladder cancer to liver in vivo. Isolated from hepatic metastasis, the line of GBC-SD/M is a subpopulation with high metastatic potential, retaining the histological properties and identification of genetic background of its parent cell line GBC-SD.

[Surgical treatment of hepatocellular carcinoma with tumor thrombus in inferior vena cava].

OBJECTIVE: To review the experience for the management of hepatocellular carcinoma with tumor thrombus in inferior vena cava. METHODS: From July 2003 to May 2005, hepatectomy combined with thrombectomy were performed on 7 cases of hepatocellular carcinoma with tumor thrombus in inferior vena cava. In order to remove the tumor thrombus in inferior vena cava, total hepatic vascular exclusion were adopted on all cases to control the blood flow of IVC. According to the position of extension of tumor thrombus, 5 different procedures were adopted in the cases to control the suprahepatic IVC and extract the tumor thrombus out of IVC and atrium. Procedure 1: Median sternotomy, extracorporeal bypass, cardiac arrest, incision on right atrium and IVC were performed on 1 case for thrombectomy. Procedure 2: Median sternotomy, extracorporeal bypass without cardiac arrest, incision on IVC and (or without) incision on right atrium were performed on 2 cases for thrombectomy. Procedure 3: Abdominal approach to control intrapericardial IVC through an incision on diaphragm was performed on 1 case for thrombectomy. Procedure 4: Abdominal approach to control suprahepatic IVC above diaphragm through a small incision made on vena cava foramen for thrombectomy was performed on 1 case. Procedure 5: Abdominal approaches to control suprahepatic IVC below diaphragm for thrombectomy were performed on 2 cases. RESULTS: All operations were successfully performed. The postoperative complications included pleural effusion in 1 case, subphrenic fluid collection in 1 case and wound infection in 1 case. The average survival time of 7 cases was 9.8 month. The longest survival time was 26 months. CONCLUSION: Hepatectomy and thrombectomy can be safely performed on the case of HCC combined with tumor thrombus in IVC. Surgical treatment can relieve the patient from the risk of sudden death caused by heart failure and pulmonary.

[Comparison of wound healing after pancreaticojejunostomy with three anastomotic methods in piglets].

OBJECTIVE: To evaluate wound healing after pancreaticojejunostomy of three anastomotic methods. METHODS: Fifty-four domestic piglets were divided into three groups according to the types of anastomoses: group of end-to-end pancreaticojejunal invagination (EE group), group of binding pancreaticojejunostomy (BP group) and group of inkwell pancreaticojejunostomy (IP group). Bursting pressure, breaking strength and histopathological findings of anastomosis were assessed on operative day and on the 5th and 10th day after operation. RESULTS: Bursting pressure was (67+/-8) mm Hg, (96+/-11) mm Hg and (131+/-9) mm Hg in EE group on day 0, 5 and 10; and (140+/-8) mm Hg, (179+/-10) mm Hg and (269+/-13) mm Hg in BP group; and (102+/-10) mm Hg, (171+/-18) mm Hg and (254+/-24) mm Hg in IP group. Compare to EE group, bursting pressure of BP group and IP group were all increased with significant differences (P<0.05). Another significant difference was observed between BP group and IP group after anastomoses on operative day. Breaking strength was (4.6+/-0.6) N, (5.8+/-0.5) N and (7.1+/-0.6) N in EE group on 0 d, 5 d and 10 d; and (4.5+/-0.4) N, (6.6+/-0.4) N and (10.0+/-0.6) N in BP group; and (4.6+/-0.3) N, (6.5+/-0.4) N and (9.1+/-0.9) N in IP group. A similar value of anastomoses was shown in BP group and IP group on day 0, day 5 and day 10, but significant increase was demonstrated compared to EE group on day 5 and 10. Anastomotic site was well repaired by connective tissue and the cut surface of pancreatic stump was covered by mucosal epithelium in BP group and IP group on day 10, but the cut surface was incompletely repaired by granulation tissue and no regeneration of the epithelium was found in EE group. CONCLUSIONS: Wound healing of binding pancreaticojejunostomy and inkwell pancreaticojejunostomy is more rapid and better than end-to-end pancreaticojejunal invagination, but breaking strength of inkwell pancreaticojejunostomy is weaker than binding pancreaticojejunostomy.

[Retrograde caudate lobectomy in 7 cases].

OBJECTIVE: To explore the clinical value and significance of retrograde caudate lobectomy. METHODS: From December 2003 to January 2005, 7 patients underwent retrograde caudate lobectomy in which division and ligation of short hepatic veins were carried out at the final stage of the procedure in stead of at the initial stage. RESULTS: The procedures were carried out smoothly with no operative death in all the 7 cases including isolated complete caudate lobectomy in 4 cases, isolated partial caudate lobectomy in 1 case, combined right half liver resection in 2 case. The average operation time, blood loss and length of stay after operation was (273 +/- 44) min, (1114 +/- 241) ml (800-1500 ml) and 16 days respectively. Complications including pleural effusion and ascites in 1 case respectively were fully recovered. During the follow-up, 1 patient died at 6 months for tumor recurrence in lung and the remaining 6 patients are alive at the follow-up of 5 to 16 months. CONCLUSIONS: Retrograde caudate lobectomy is a new procedure suitable for those caudate neoplasms which are adhering to or infiltrating to IVC or too big to move side by side. The application of this technique can converse certain kind of caudate lobe tumor from non-resectable to resectable resulting in widening the indication.

[Reverse first dorsal metatarsal artery island flap for coverage of donor defect of great toe after free lateral pulp flap transfer].

OBJECTIVE: To employ reverse first dorsal metatarsal artery island flap to recover the donor site of great toe after free lateral pulp flap transfer, and decrease the morbidity of microsurgical donor site. METHODS: From February 2000 to June 2004, 12 cases of soft tissue defect in thumb and finger were treated by free lateral pulp flap of great toe transplantation, and that the secondary defect of great toes were repaired by reverse first dorsal metatarsal artery flap according to the anatomical communicating between first dorsal metatarsal artery and plantar metatarsal artery. The donor exposure of dorsal pedis were straightly sutured. RESULTS: All the lateral pulp flaps of great toe and the reversed first dorsal metatarsal flaps survived uneventfully with desirable appearance and sensation. In thumb and finger defect reconstructed with lateral pulp flaps of the great toe, there was in mean static two-point discrimination of 6 mm at 10 month follow-up, that was 10 mm in reverse flaps. CONCLUSIONS: It is an instructive and practical creation to adopt the reverse first dorsal metatarsal artery flap to recover the raw surface in the lateral aspect of the great toe, which minimizes the morbidity at the donor site and gains the twin-win results.