Objective:To observe the effects of preventative moxibustion on analgesia,substance P(SP),prostaglandin(PG)F2α and PGE2 in rats with dysmenorrhea due to cold-dampness stagnation,and to explore the analgesic mechanism. Methods:Sixty-four female Wistar non-pregnant rats were randomly divided into a blank group,a model group,a Western medicine group,and a preventative moxibustion group,with 16 rats in each group.Eight qualified diestrus rats were selected from each group.Except for the blank group,the other three groups established models of dysmenorrhea due to cold-dampness stagnation using an ice water bath combined with estradiol benzoate and oxytocin.On the 8th day after modeling,the preventative moxibustion group was treated with gentle moxibustion at Shenque(CV8)and Guanyuan(CV4),and the Western medicine group was given ibuprofen solution for 4 consecutive days.On the 11th day,the intervention groups(i.e.the Western medicine group and the preventative moxibustion group)were treated once again after being injected with oxytocin.The writhing score and the pain threshold of rats were determined;the serum levels of brain-derived neurotrophic factor(BDNF),SP,PGF2α,and PGE2 were measured;the mRNA and protein expression levels of BDNF and its receptor tropomyosin receptor kinase B(TrkB)in the spinal dorsal horn and hypothalamus were detected. Results:Compared with the blank group,the writhing score increased(P<0.01),the pain threshold decreased(P<0.01),the serum levels of BDNF,SP,and PGF2α increased(P<0.01),while the PGE2 decreased(P<0.01);the protein and mRNA expression levels of BDNF and TrkB in the spinal dorsal horn and hypothalamus increased(P<0.01)in the model group.Compared with the model group,the writhing score decreased,the pain threshold increased,the serum BDNF,SP,and PGF2α levels decreased significantly,the serum PGE2 level increased,and the protein and mRNA expression levels of BDNF and TrkB in the spinal dorsal horn and hypothalamus decreased significantly in the preventative moxibustion group and the Western medicine group,while the inter-group differences were significant(P<0.01).Compared with the Western medicine group,the writhing score decreased,the pain threshold increased,the serum BDNF,SP,and PGF2α,levels decreased,the serum PGE2 level increased,and the protein and mRNA expression levels of BDNF and TrkB in the spinal dorsal horn and hypothalamus in the preventative moxibustion group decreased significantly,while the inter-group differences were significant(P<0.05 or P<0.01). Conclusion:Preventative moxibustion at Shenque(CV8)and Guanyuan(CV4)can improve the pain sensitization state of rats with dysmenorrhea due to cold-dampness stagnation,down-regulate the mRNA and protein expression levels of BDNF and TrkB in the spinal dorsal horn and hypothalamus;regulation of the serum SP,PGF2α,and PGE2 levels may be part of the mechanism.

Background/Aims: The shortage of donor liver hinders the development of liver transplantation. This study aimed to clarify the poor outcomes of functionally marginal liver grafts (FMLs) and provide evidence for the improvement of ischemia-free liver transplantation (IFLT) after FML transplantation. Methods: Propensity score matching was used to control for confounding factors. The outcomes of the control group and FML group were compared to demonstrate the negative impact of FMLs on liver transplantation patients. We compared the clinical improvements of the different surgical types. To elucidate the underlying mechanism, we conducted bioinformatic analysis based on transcriptome and single-cell profiles. Results: FMLs had a significantly greater hazard ratio (HR: 1.969, P=0.018) than did other marginal livers. A worse 90-day survival (Mortality: 12.3% vs. 5.0%, P=0.007) was observed in patients who underwent FML transplantation. Patients who received FMLs had a significant improvement in overall survival after IFLT (Mortality: 10.4% vs 31.3%, P=0.006). Pyroptosis and inflammation were inhibited in patients who underwent IFLT. The infiltration of natural killer cells was lower in liver grafts from these patients. Bulk transcriptome profiles revealed a positive relationship between IL-32 and Caspase 1 (R=0.73, P=0.01) and between IL-32 and Gasdermin D (R=0.84, P=0.0012). Conclusions FML is a more important negative prognostic parameter than other marginal liver parameters. IFLT might ameliorate liver injury in FMLs by inhibiting the infiltration of NK cells, consequently leading to the abortion of IL-32, which drives pyroptosis in monocytes and macrophages.

Epithelioid Hemangioendotheliomais a rare, low-grade malignant vascular tumour. It’scalled hepatic epithelioid hemangioendothelioma(HEHE), when it occurs in liver. It can be metastatic and postoperative recurrence. There are few cases have been reported in the literature at home and abroad because of its rarity. The treatment of HEHE is also controversial. With the continuous improvement of surgical techniques of liver transplantation, it is increasingly applied to treat liver failure patients caused by HEHE. Our paper reviews the literature on disease characteristics of HEHE, and liver transplantation for HEHE indications, immunotherapy and prognosis, to illustrate the status and progress of liver transplantation for HEHE.

Objective To investigate the effect of expression of Cullin 4B (CUL4B) on the prognosis of patients after liver transplantation for hepatocellular carcinoma (HCC).Methods The retrospective case-control study was conducted.The clinicopathological data of 79 patients who underwent liver transplantation for HCC in the First Affiliated Hospital of Sun Yat-sen University between January 1,2014 and June 30,2015 were collected.The specimens of HCC tissues were collected and embedded in paraffin,and then were detected by immunohistochemistry staining.Observation indicators:(1) expression of CUL4B in HCC tissues;(2) follow-up and survival;(3) prognostic factors analysis after liver transplantation;(4) association between expression of CUL4B and recurrence and metastasis of tumor after liver transplantation.Follow-up using outpatient examination and telephone interview was performed to detect tumor recurrence or metastasis and survival up to June 2018.Measurement data with normal distribution were represented as (x)±s.The comparison between groups of count data was done using the chi-square test.The survival curve drawn using the Kaplan-Meier method,and the survival analysis was done by Log-rank test.The univariate and multivariate analysis were respectively done using the COX regression model.The association analysis was done using the Pearson test.Results (1) Expression of CUL4B in HCC tissues:immunohistochemistry staining showed that CUL4B was mainly expressed in the cytoplasm,with a powerful brownish-yellow staining.The high expression and low expression of CUL4B in HCC tissues were detected in 64 and 15 patients,respectively.(2) Follow-up and survival:79 patients were followed up for 38-56 months,with an average time of 46 months.During the follow-up,37 patients had no tumor recurrence and 42 had tumor recurrence (32 with tumor extrahepatic metastasis and 10 with intrahepatic metastasis);36 had survival and 43 died;the 1-and 3-year overall survival rates were respectively 86.84％ and 63.25％,and 1-and 3-year tumorfree survival rates were respectively 62.31％ and 51.27％.(3) Prognostic factors analysis after liver transplantation:① Results of univariate analysis showed that preoperative alpha-fetoprotein (AFP),Child-Pugh score,maximum tumour dimension,capsular invasion,intravascular tumor thrombus,Edmonson pathological grading and expression of CUL4B were related factors affecting the 3-year overall survival rate of patients after liver transplantation for HCC [Hazard Ratio (HR) =2.17,3.36,3.66,2.43,2.19,3.36,2.84,95％ confidence interval(CI):1.17-4.04,1.53-7.42,2.10-6.42,1.33-4.17,1.08-9.04,1.58-7.59,1.17-6.32,P＜ 0.05].The preoperative alpha-fetoprotein (AFP),Child-Pugh score,maximum tumour dimension,capsular invasion,intravascular tumor thrombus,Edmonson pathological grading and expression of CUL4B were related factors affecting the 3-year tumor-free survival rate of patients after liver transplantation for HCC (HR =2.06,3.72,3.16,2.36,2.83,3.21,1.69,95％CI:1.34-4.85,1.72-8.63,1.79-7.31,1.46-4.86,1.19-8.63,1.19-7.92,1.06-4.87,P＜0.05).② Results of multivariate analysis showed that maximum tumour dimension,intravascular tumor thrombus and expression of CUL4B were independent factors affecting the 3-year overall survival rate of patients after liver transplantation for HCC [Odds ratio(OR) =3.43,3.69,2.81,95％CI:1.16-6.02,1.96-9.38,1.04-9.63,P＜0.05].The maximum tumour dimension,intravascular tumor thrombus and expression of CUL4B were independent factors affecting the 3-year tumor-free survival rate of patients after liver transplantation for HCC (OR=2.25,4.72,2.74,95％C1:1.16-4.02,1.98-9.47,1.03-7.10,P＜ 0.05).The 3-year overall survival rate in patients with high-and low-expressions of CUL4B was respectively 66.7％ and 32.8％,with a statistically significant difference (x2 =5.69,P＜0.05).The 3-year tumor-free survival rate in patients with high-and low-expressions of CUL4B was respectively 73.3％ and 18.6％,with a statistically significant difference (x2 =4.63,P＜0.05).(4) Association between expression of CUL4B and recurrence and metastasis of tumor after liver transplantation:results of Pearson test showed that expression of CUL4B was significantly associated with HCC recurrence and metastasis after liver transplantation (r =0.62,P＜0.05).The further analysis showed that expression of CUL4B was significantly associated with extrahepatic metastasis after liver transplantation (r=0.84,P ＜ 0.05).Conclusion The expression of CUL4B is associated with HCC recurrence after liver transplantation,and it can be as a predictor for HCC recurrence and distant metastasis after liver transplantation.

Objective: To observe the effects of moxibustion at different times on prostaglandin E2 (PGE2), prostaglandin F2α (PGF2α) and arginine vasopressin (AVP), in the uterine tissues of rats with dysmenorrhea due to cold-dampness retention, and to explore the differences and possible mechanisms of moxibustion at different times in easing pain in dysmenorrhea due to cold-dampness retention. Methods: Forty-three female Wistar rats were randomly divided into a blank control group (n=7), a model group (n=9), a pre-moxibustion group (n=9), an immediate-moxibustion group (n=9) and a pre-moxibustion plus immediate-moxibustion group (n=9). Rat models of primary dysmenorrhea due to cold-dampness retention were established using (0±1) ℃ ice water immersion method combined with injection of estradiol benzoate for 10 d, followed by injection of oxytocin on the 11th day. Rats in the 3 intervention groups received moxibustion to Shenque (CV 8) and Guanyuan (CV 4), 10 min for each acupoint, once a day. Rats in pre-moxibustion group were given mild moxibustion, beginning on the 8th day during modeling, for 3 continuous days; rats in immediate-moxibustion group were given one time mild moxibustion, immediately after injection of oxytocin on the 11th day during modeling; rats in pre-moxibustion plus immediate-moxibustion group were given mild moxibustion, beginning on the 8th day during modeling till immediately after injection of oxytocin on the 11th day during modeling, for 4 continuous days. The level of PGF2α in the rat uterine tissues was measured by enzyme-linked immunosorbent assay (ELISA), and the levels of PGE2 and AVP in rat uterine tissues were measured by radioimmunoassay. Results: Compared with the blank control group, the levels of PGF2α and AVP, the PGF2α/PGE2 ratio in the model group were significantly increased (P<0.01), and the PGE2 level was significantly decreased (P<0.01) in the rat uterine tissues in the model group. Compared with the model group, the writhing latency was significantly prolonged, the writhing number and the total writhing score were all decreased in the pre-moxibustion group, the immediate-moxibustion group and the pre-moxibustion plus immediate-moxibustion group (all P<0.01); the levels of PGF2α and AVP, and the PGF2α/PGE2 ratio were all significantly decreased (P<0.05, P<0.01), and the PGE2 level was significantly increased (P<0.01) in the rat uterine tissues of the 3 treatment groups. Compared with the pre-moxibustion group, the writhing number and the total writhing score were all decreased in the immediate-moxibustion group and the pre-moxibustion plus immediate-moxibustion group (all P<0.01), the writhing latency was significantly prolonged in the pre-moxibustion plus immediate-moxibustion group(P<0.01); the levels of PGF2α and PGF2α/PGE2 ratio were significantly decreased (P<0.05, P<0.01), and the PGE2 level was significantly increased (P<0.01) in rat uterine tissues in the immediate-moxibustion group and the pre-moxibustion plus immediate-moxibustion group. Compared with the immediate-moxibustion group, the writhing latency was significantly prolonged and the writhing number was decreased (all P<0.05), and the total writhing score was decreased (P<0.01) in the pre-moxibustion plus immediate-moxibustion group; the PGF2α level and the PGF2α/PGE2 ratio were significantly decreased (P<0.01), and the PGE2 level was significantly increased (P<0.01) in rat uterine tissues in the pre-moxibustion plus immediate-moxibustion group. Conclusion: Moxibustion at different times all can produce obvious analgesic effects on dysmenorrhea due to cold-dampness retention in rats, and pre-moxibustion plus immediate-moxibustion ranks the top. The mechanism of this analgesic effect may be via the regulation of abnormal PGF2α, PGE2 and AVP levels, to effectively inhibit the spastic contraction of uterine smooth muscle in dysmenorrhea rat, thereby improving the ischemia and hypoxia in uterus.

OBJECTIVE:To explore the repairing effect and mechanism of Huazhuo jiedu yukui formula on the mucosal dam-age of rats with ulcerative colitis (UC). METHODS:72 rats were randomly divided into blank group,model group,mesalamine group(positive control,0.3 g/kg),Huazhuo jiedu yukui formula low-dose,medium-dose,high-dose groups(5,10,20 g/kg),12 in each group. Except for blank group,other groups were induced UC model. After 4 d of modeling,rats in administration groups received related medicines,ig,blank group and model group received normal saline,ig,once a day,for 2 weeks. After administra-tion,disease activity index(DAI)and mucosal damage index(CMDI)of rats were observed;IL-8,TNF-α contents in serum and cyclooxygenase 2 (COX-2) expression in colon tissue were detected. RESULTS:Compared with blank group,DAI and CMDI scores in model group were increased;IL-8,TNF-α contents in serum were increased;COX-2 expression in colon tissue was en-hanced,with statistical significances (P<0.05). Compared with model group,DAI and CMDI scores in model group were de-creased;IL-8,TNF-α contents in serum were reduced;COX-2 expression in colon tissue was weakened,with statistical signifi-cances (P<0.05). Compared with mesalamine group, the above-mentioned indicators in Huazhuo jiedu yukui formula medi-um-dose,high-dose groups changed more obviously (P<0.05),and effects of Huazhuo jiedu yukui formula showed significant dose-effect relationship. CONCLUSIONS:Huazhuo jiedu yukui formula can improve disease activity status of UC rats and reduce colon mucosal damage,and the effects of medium-dose and high-dose Huazhuo jiedu yukui formula were better than mesalamine, which may be related to reducing inflammatory reaction and inhibiting COX-2 expression in colon tissue.

Objective To investigate the expression of Kuppel-like factor 2( KLF2 )after focal cerebral ischemia-reperfusion( I/R)injury in rats and the intervention effect of nuclear factor kappa B ( NF-κB)inhibitor. Methods Sixty healthy male SD rats were randomly divided into a sham operation group,an I/R group,and a NF-κB inhibitor group( n=20 in each group). A focal cerebral I/R model was induced by the intraluminal suture method,and NF-κB inhibitor( pyrrolidinedithio carbamate,PDTC)was given to intervene. The observation time points were 6,12,24,and 48 hours after I/R. Reverse transcription-polymerase chain reaction(PCR)and Western blot were used to measure KLF2 mRNA and protein expression in ischemic brain tissue. Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of serum tumor necrosis factorα( TNF-α),and they were compared among groups. Results Compared with the sham operation group,the expression levels of KLF2 mRNA and protein in I/R group in the ischemic brain tissue at each time point were averagely decreased( the relative expression levels of KLF2 mRNA:0. 46 ± 0. 03 vs. 0. 82 ± 0. 04,0. 30 ± 0. 04 vs. 0. 78 ± 0. 05,0. 18 ± 0. 04 vs. 0. 76 ± 0. 02,0. 26 ± 0. 02 vs. 0. 81 ± 0. 04,respectively;the relative expression levels of KLF2 protein:0. 46 ± 0. 04 vs. 0. 80 ± 0. 02,0. 30 ± 0. 02 vs. 0. 79 ± 0. 02,0. 15 ± 0. 02 vs. 0. 77 ± 0. 01,0. 24 ± 0. 01 vs. 0. 79 ± 0. 02,respectively). They reached the lowest values at 24 hours after I/R,while the serum TNF-αlevels were increased. There were significant differences(all P<0. 05). After giving NF-κB inhibitor PDTC,the expression levels of KLF2 mRNA and protein at 6,12,24,and 48 hours after I/R were upregulated differently compared with the I/R group. The relative expression levels of KLF2 mRNA were 0. 61 ± 0. 04,0. 44 ± 0. 03,0. 34 ± 0. 02,and 0. 43 ± 0. 04, respectively. Those of KLF2 protein were 0. 60 ± 0. 02,0. 43 ± 0. 02,0. 33 ± 0. 01,and 0. 44 ± 0. 03, respectively,while the levels of TNF-αwere decreased. There were significant differences(all P<0. 05). There was a negative correlation between the KLF2 mRNA levels and the serum TNF-αlevels at each time point in the I/R group and the PDTC group( r= —0. 728 ,P<0. 05 ). Conclusions The expression levels of KLF2 mRNA in brain tissue are decreased after I/R,and it is negatively correlated with the serum TNF-α levels. It may be involved in the pathological process of I/R by NF-κB pathway mediated inflammatory reaction.

Objective To investigate protective effects of thrombopoietin ( TPO) on cerebral model control in rats and associated signal transduction pathway. Methods Thread embolism was performed to generate cerebral ischemia-reperfusion rat model. Eighty male SD rats were randomly divided into sham operation group, model control group, TPO group, TPO and Janus kinase 2 ( JAK2 ) kinase inhibitor ( AG490 ) group. Before 30 min of ischemia-reperfusion, TPO group was given TPO (5 μg·kg-1) by intraperitoneal injection, TPO + AG490 group was given TPO (5 μg·kg-1) before 30 min of ischemia reperfusion, then given AG490 (8 μg·kg-1), and model control group were given the same dose of 0. 9% sodium chloride solution. The observation time points were 6, 12, 24, and 48 h after ischemia reperfusion. Immunohistochemical staining and Western blotting were used to measure the protein levels of Bcl-2, JAK2 and signal transducer & activator of transcription (STAT3). TdT-mediated dUTP nick end labeling (TUNEL) was used to detect apoptosis. Results Compared with model control group, the number of apoptotic cells were significantly reduced [(67. 50±9. 37) vs. (40. 20±7. 47)], the expression levels of Bcl-2, JAK2 and STAT3 protein were significantly increased [(35. 40±7. 39) vs. (78. 70±9. 75);(35. 68±6. 75) vs. (62.35±7.53); (25.40±9.45) vs.(55.36±9.69), respectively] 24 h after ischeia reperfusion in the TPO group (all P<0. 05). Compared with the TPO group, the Bcl-2, JAK2 and STAT3 protein levels were significantly decreased in TPO and AG490 group [(78. 70±9. 75) vs. (55. 40±9. 35);(62. 35±7. 53) vs. (40. 68±5. 89); (55. 36±9. 69) vs. (30. 40±9. 39), respetively], and the number of apoptotic cells was significantly increased [(40. 20±7. 47) vs. (55. 23±7. 65)] (all P<0. 05). Conclusion TPO can inhibit cell apoptosis after ischemia-reperfusion injury, the mechanism might be related to the activation of JAK2/STAT3 signal transduction pathway through raising the expression of Bcl-2 gene.

OBJECTIVETo determine whether the miRNA expression profile in peripheral blood mononuclear cells (PBMCs) differs between liver transplant recipients with long-term stable survival and those with acute rejection.

METHODSTwenty-nine liver transplant recipients with long-term stable survival (STA) group, 10 recipients with acute rejection (RJ group), and 17 healthy subjects (control group) were recruited for genome-wide microarray analysis of miRNA expressions in the PBMCs. The differentially expressed miRNAs among the 3 groups were validated by real-time PCR, and the targets of these miRNAs were predicted.

RESULTSCompared with the RJ group, the STA group showed down-regulation of 13 miRNAs in the PBMCs. Of these down-regulated miRNAs, miRNA-18b, miRNA-340 and miRNA-106b were validated by real-time PCR, and the latter two miRNAs were predicted to target the TGF-β pathway.

CONCLUSIONSThe differentially expressed miRNAs in liver transplant recipients with long-term stable survival, namely miRNA-18b, miRNA-340 and miRNA-106b, can be potential clinical biomarkers to predict the outcomes of liver transplantation.

Biomarkers ; metabolism ; Case-Control Studies ; Down-Regulation ; Graft Rejection ; Humans ; Leukocytes, Mononuclear ; metabolism ; Liver Transplantation ; MicroRNAs ; metabolism ; Oligonucleotide Array Sequence Analysis ; Real-Time Polymerase Chain Reaction ; Survival Rate ; Transforming Growth Factor beta

Objective To observe the influencing factors of polysomnography (PSG) monitoring and to evaluate the efficacy of preventive measures.Methods We selected 205 patients with sleep apnea syndrome (SAS) who accepted PSG from May 2011 to April 2012 in our hospital as the intervention group.They were administered preventive measures,including psychological counseling,intensive grease dispelling of patient skin,arrangement of sensor and electrode.One hundred and fifty-two SAS patients who were administered PSG from May 2010 to April 2011 were selected as the control group.The monitoring successful rates between groups were compared and influencing factors were investigated.Results The monitoring successful rates of control group and the intervention group were 92.1％ and 97.1％ respectively; there was a statistically significant difference between the successful rates of the two groups (x2 =4.499,P ＜ 0.05).The main causes for unsuccessful monitoring were difficulty falling asleep,electrode distortion and high impedance of electrode.Conclusion PSG monitoring has a complex operating procedure and is time-consuming and effective preventive measures can improve the successful rate of PSG.