Objective To investigate the value of intravital fluorescence microscopy in the observation of the changes of hepatic microcirculation in the rat model with hepatic cirrhosis and portal hypertension.Methods Seventy male SD rats were selected.According to the random number table,40 SD rats were randomly divided into the sham operation group,bile duct ligation (BDL) 2 weeks group,4 weeks group and 6 weeks group,there were 10 rats in each group,and the hepatic microcirculation of the rats was observed with intravital fluorescence microscope; the remaing 30 SD rats were randomly divided into the normal saline (NS) group,endothelin-1 (ET-1) group and the S-nitrosoglutathion (GSNO) group at 4 weeks later after the establishment of BDL model.The changes of hepatic microcirculation of the 3 groups were observed.All data were analyzed using the one-way analysis of variance (ANOVA) or paired samples t test.Results Nine rats died in the BDL model groups,and the survival rate was 85.0％ (51/60).All rats in the sham operation group were survived.The hepatic sinusoid diameters were decreased as time passed by.The hepatic sinusoid diameters of the BDL 2 weeks group,4 weeks group and 6 weeks group were (13.6 ± 1.0) μm,(8.8 ± 0.7) μm and (8.0 ± 0.5) μm,respectively,which were significantly shorter than (17.4 ± 1.0) μm of the sham operation group (t =5.86,18.24,15.57,P ＜ 0.05).The hepatic sinusoid densities of the BDL 2 weeks group,4 weeks group and 6 weeks group were (6.8 ±0.8)/ 200 μm,(4.3 ± 1.8)/200 μm and (4.0 ± 1.2)/200 μm,which were significandy lesser than (8.8 ± 0.5)/200 μm (t =3.25,2.77,2.12,P ＜ 0.05).At 15 minutes after injection of NS,the hepatic sinusoid diameter of the NS group was (7.2 ± 1.2) μm,which was significantly different from (6.9 ± 0.5) μm before injection of NS (t =0.89,P ＞ 0.05) ; the hepatic sinusoid density of the NS group before and after injection of NS were (6.6 ± 0.4) / 200 μm and (6.8 ± 1.4)/200 μm,with no significant difference(t =1.12,P ＞0.05).At 15 minutes after injection of ET-1,the hepatic sinusoid diameter of the ET-1 group was (5.4 ±0.5) μm,which was significantly different from (7.9 ± 0.6) μm before injection of ET-1 (t =7.39,P ＜ 0.05) ; the hepatic sinusoid density of the ET-1 group before and after ET-1 injection were (5.8 ± 1.2)/200 μm and (5.4 ± 1.8)/200 μm,with no significant difference(t =0.84,P ＞0.05).At 15 minutes after injection of the GSNO,the hepatic sinusoid diameter of the GSNO group was (11.4 ± 1.3) μm,which was significantly different from (7.5 ± 1.7) μm before injection of GSNO (t =5.95,P ＜ 0.05) ; the hepatic sinusoid density of the GSNO group before and after GSNO injection were(5.6 ± 0.8)/200 μm and (6.4 ± 1.6)/200 μm,with no significant difference (t =0.54,P ＞ 0.05).Conclusions The changes of hepatic microcirculation observed under intravital fluorescence microscope could reflect the progress of hepatic cirrhosis,and the changes of hepatic sinusoid diameters caused by drugs could be dynamically monitored under the intravital fluorescence microscope.

Objective To investigate the role of autophagy in podocyte damage,and the intracellular mechanism of autophagy activation through passive Heymann nephritis (PHN) animal model.Methods Male Sprague-Dawley rats (n=40) were studied on day 0,2,4,7,and 21 after induction of PHN by injection of anti-Fx1A.Podocyte morphology and autophagosomes were observed by transmission electron microscopy.Podocyte numerical density was estimated by Weibel-Gomez =method.Cell apoptosis was detected by TUNEL assay and caspase-3 immunohistochemical staining.Expressions of autophagy markers and endoplasmic reticulum stress (ERS)-associated proteins were analyzed by Western blotting.Results (1) In PHN rats,immunohistochemical staining showed that C5b-9 deposited along glomerular basement membrane on day 4 to day 21.Small subepithelial electron -dense deposits and a part of foot process fusion were detected in the glomerulus of PHN rats on day 4 by transmission electron microscope,and podocyte damage was aggravated on day 21.Furthermore,compared with control,the urinary protein levels of PHN rats began to increase on day 3,and reached the top on day 21 [(50.6±6.0) mg/24 h].(2) The number of podocytes significantly decreased in PHN rats compared with control group on day 21(P ＜ 0.05).(3) In PHN rats,apoptotic podocytes were found by caspase-3 immunohistochemical staining and TUNEL assay on day 21.(4) The expression of autophagy marker LC3 Ⅱ was markedly increased on day 7 and 21,but down-regulated on day 21 compared with day 7.Moreover,accumulated autophagosomes in podocytes were detected on day 7 and 21 by transmission electron microscope.(5) The level of GRP78 was significantly increased on day 2 and 7 but reduced to baseline on day 21.At the same time,the downstream pathways (ATF6α,p-PERK and p-JNK) of unfolded protein response were also up-regulated in the early process of PHN and down-regulated later.Conclusions Autophagy is an important way to protect against immunemediated podocyte injury in membranous nephropathy.Autophagy activation is mainly related to endoplasmic reticulum stress induced by complement attack.This provides an important basis for a thorough understanding of the role of autophagy in the process of podocyte damage and the pathogenesis of membranous nephropathy.

Objective To observe any differences in the morphology of the lumbar multifidus (LM) muscles between normal subjects and persons with chronic lumbar disc herniation (CLDH).Methods Thirty-two persons with CLDH and thirty-five matched,healthy controls performed various maneuvers in prone lying.Ultrasound imaging was used to measure the antero-posterior diameter (A-P),lateral diameter (Lat),cross sectional area (CSA) and flattening ratio (FR) of the bilateral LM muscles at the L4/5 level.Results In the control subjects,A-P,Lat,CSA and FR were not significantly different between the left and right sides.In those with CLDH,A-P,Lat and CSA were significantly smaller on the painful side than on the no-pain side,and this difference showed asymmetry between the two sides.Average A-P,Lat,FR and muscle CSA were all significantly smaller among the CLDH subjects on both sides compared with the controls.Conclusion CLDH induces asymmetric local atrophy of the bilateral multifidus muscles which can be detected and measured using ultrasound.

TGR5,expressing in many tissue cells,is a kind of bile acid membrane receptor and participates in a variety of metabolic and immune diseases.Activated TGR5 can keep the metabolism system in a steady state by mediating the metabolism of bile acid,lipid,and blood sugar,reducing insulin resistance and increasing the body's energy consumption; TGR5 could regulate the immune responses of mononuclear cell and Kupffer cell in the liver.For example,it can regulate the adaptive immune response by inhibiting the expression and release of inflammatory cytokines in Kupffer cells,and regulating the differentiation and maturation of dendritic cells.This review mainly focused on the function of TGR5 in liver metabolism and immune and further explored the related mechanism,as well as its clinical significance in related liver diseases.

Objective To compare the applicability of the Beijing Version of the Montreal Cognitive Assessment (MoCA) and the Mini Mental State Examination (MMSE) in screening for cognitive impairment in patients with acute ischemic stroke for 2-3 weeks.Methods MoCA and MMSE were conducted in 201 patients with acute ischemic stroke within 2 to 3 weeks after the onset of stroke.With MoCA＜23 and MMSE ＜26 as the cut off value,we assessed the clinic effect of the MoCA and MMSE and explored the correlation between two instruments.Results The average scores of MoCA and MMSE scale were (20.5±4.3) and (25.4±3.5) points.The prevalence of cognitive impairment evaluated with MoCA and MMSE were 57.2％and 43.3％,respectively.MoCA showed significant correlation with MMSE score (Pearson's correlation coefficient=0.833,P＜0.001),and an agreement with Kappa values of 0.532 (P＜0.01) in screening for cognitive impairment.Conclusions The prevalence of cognitive impairment assessed with MoCA is higher than that of with MMSE when using MoCA＜23 and MMSE＜26 as the cut off values.Both instruments show a good agreement for screening cognitive impairment in acute ischemic stroke within 2 to 3 weeks following the disease onset.

Objective:To evalte a novel laparoscopic splenic artery ligation plus devascularization (LSALD) vs. laparoscopic splenectomy and devascularization (LSD) for the treatment of portal hypertention. Methods:From Jan 2014 to Dec 2019, 50 patients undergoing LSALD and 30 patients receiving LSD . We compared the safety and feasibility between LSALD and LSD groups by analyzing the patients′ blood routine, liver function before and after operation, intraoperative condition, postoperative recovery and prognosis.Results:The operation time[(181±72)min vs.(284±72)min , t=-6.205, P<0.01], intraoperative blood loss[(100±50)ml vs.( 700±86 ml), t=-5.166, P<0.01]and blood transfusion rate (28% vs.67%, χ 2=11.471, P<0.01)in LSALD group were significantly more favorite than those in LSD group ( P<0.05). The postoperative exhaust in the LSALD group was earlier than that in the LSD group (2 d vs.3 d, Z=2.361, P<0.05) though the WBC and blood platelet count was higher in LSD group ( P<0.05). Portal vein thrombosis occurred in 10 cases in LSD group and 6 cases in LSALD group (χ 2=5.757, P<0.05). Conclusion:Compared with laparoscopic splenectomy combined with periesophagogastric devascularization, laparoscopic splenic artery ligation combined with periesophagogastric devascularization is less traumatic, helping quick recovery and lower rate of post-op portal vein thrombosis.

Objective: To investigate the clinical features and reliable diagnostic method in HIV/AIDS patients with smear negative pulmonary tuberculosis (TB). Methods: Clinical data of 112 HIV/AIDS patients complicated with smear negative pulmonary TB who were treated in our hospital from January 2013 to September 2015 were retrospectively analyzed. These clinical data includeded clinical symptom, blood routine test, blood biochemistry, T lymphocyte subsets classification, sputum acid-fast bacillus smear, mycobacterium tuberculosis culture, purified protein derivatives tuberculin (PPD) test, interferon gamma-release assay for Mycobacterium tuberculosis (T-SPOT.TB), TB-DNA and chest computed tomography (CT). Diagnostic specificity and sensitivity of these parameters were analyzed. Results: No specific clinical manifestation of these patients was identified. The chest CT feature was also atypical. The positive rates including T-SPOT, TB, TB-DNA and PPD test were all low. The positive rates of T-SPOT.TB and PPD test in patients with a CD4+ cell count >200 cells/μl was significantly higher than that of patients with a CD4+ cell count ≤50 cells/μl and 51≤CD4≤200 cells/μl (P<0.01). Conclusions The clinical feature of smear negative pulmonary TB in HIV/AIDS patients is atypical. For a definite diagnosis, a comprehensive analysis with clinical manifestations, laboratory test, imaging examination and etiologic detection is required.

Objective:To explore the effect of grief counseling for relieving donor family′s grief. Methods:From September 2012 to February 2015, 180 families of potential organ donors, who met the class III standard of China, were invited to participate in this study. The grief score was evaluated using questionnaire before and after grief counseling. Results:All of 180 potential organ donor′s families had different level of sadness. The grief was significantly reduced after grief counseling and the score was significantly lower than before ( P<0 . 05 ) . Sixty-five cases agreed to donate organ and 60 cases succeed. Conclusion:Grief counseling for potential organ donor′s families could relieve their grief effectively. This method is beneficial for communication of organ donation and pro-moting donation career of China.

Objective To explore the protective effects of quercetin on damage induced by oxidative stress and to clari?fy its molecular mechanism. Methods Chang liver cell cultures were randomly divided into control groups, H2O2 group and 3 doses of quercetin groups. Cell survival rate was detected with MTT. Cell apoptotic rate was measured by FACS(Fluores?cence-activated cell sorting). Intracellular reactive oxygen species (ROS) level in Chang liver cells were tested by flow cy?tometer. The DCF fluorescence intensity of DCFH-DA-stained intracellular ROS was observed by fluorescence microscope. The levels of malondialdehyde (MDA), superoxide dismutase(SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) were determined in liver cells using commercial available kits. The expression of Nrf2 were detected by Western blot. Re?sults Compared with control, cell survival rate and levels of SOD, CAT and GSH-Px decreased significantly in H2O2 group (P < 0.05 ),while cell appotosis rate, content of MDA and mean fluorescence intensity(MFI) increased in H2O2 group (P <0.05). In comparison with H2O2, expression of Nrf2 protein was higher in all three quercetin treatment groups (P<0.05). Con?clusion Quercetin protected Chang liver cells from H2O2-induced oxidative stress, which may be caused by the increased ex?pressions of down stream antioxidant genes via activating the Nrf2-ARE signaling pathway.

Objective To explore the expression of pituitary tumor transforming gene (PTTG) and c-myc gene in patients with non-Hodgkin lymphoma (NHL),its relationship with NHL and its clinical significance.Methods PTTG mRNA and c-myc mRNA levels in bone marrow mononuclear cell (BMMNC) isolated from 38 NHL patients and 10 chronic lymphadenitis patients were quantified by reverse transcriptionpolymerase chain reaction (RT-PCR).Results mRNA expression of PTTG and c-myc gene in BMMNC was significantly higher in NHL patients than those in normal controls (PTTG:0.567 7±0.270 7 vs 0.071 2± 0.020 1,t =4.706,P ＜ 0.05; c-myc:0.352 6±0.185 4 vs 0.107 3±0.043 5,t =3.303,P =0.002).The expression of PTTG and c-myc gene in peripheral T cell lymphoma and diffuse large B cell lymphoma showed no significant difference (PTTG:0.556 8±0.211 3 vs 0.602 8±0.244 6,t =0.640,P =0.527; c-myc:0.350 1± 0.177 6 vs 0.361 0±0.190 2,t =0.302,P =0.765).Both expression of PTTG and c-myc were positively related to NHL clinical stage and IPI.Expression of PTTG mRNA was positively related to the expression of c-myc mRNA.Conclusion There was overexpression of PTTG and c-myc in NHL,which indicates that PTTG might be involved in tumorigenesis of NHL through direct or indirect activation of c-myc oncogene.