To observe the effect of various doses of oil of Piper longum unsaponifiable matter (OPUM) to cholesterol gallstones in experimental mice. C57BL/6 mice (n = 60) were randomly divided into 6 groups: control group, model group, OPUM (15, 30 and 60 mg x kg(-1)) group and ursodeoxycholic acid (UDCA, 60 mg x kg(-1)) group, administered for 10 weeks. The level of serum lipid and liver function enzymes were tested. The gallbladder was removed and bile was obtained by centrifugation. Next, the levels of the bile total cholesterol (TC), phospholipid (PL) and bile acid (TBA) were measured. The indicators of lipid peroxidation were determined and cholesterol saturation index (CSI) was calculated. The liver histological changes were observed by HE staining. The results showed that serum TC, TG (triglycerides) and AST (aspartate transaminase) contents, gallbladder cholesterol crystallization and CSI increased significantly (P < 0.05). In addition, the activity of SOD decreased significantly and MDA content increased significantly in liver (P < 0.05). HE staining results showed that the hepatic cord disorder and intracellular lipid droplets increased significantly. All results indicate that lithogenic diet lead to the formation of cholesterol gallstones. In OPUM (30 and 60 mg x kg(-1)) group, serum TC, TG and AST content, gallbladder cholesterol crystallization and CSI decreased significantly, the activity of SOD increased significantly and MDA content decreased significantly. HE staining results showed that OPUM can improve the morphology of liver cell, reduce the degree of hepatic cord disorders and restore the cell morphology close to normal. The cause of OPUM prevents cholesterol gallstone formation maybe due to protect the integrity of the liver cells, lower CSI, and reduce cholesterol crystal formation and hence prevent cholesterol gallstone formation.

Objective To evaluate the effect of spleno-left adrenal vein shunt for the treatment of portal hypertensive upper GI bleeding caused by portal vein cavernous transformation in children.Methods Spleno-left adrenal vein shunt was performed in 8 children with portal hypertension due to cavernous transformation.The clinical data was reviewed.Results Portal vein pressure decreased significantly from (30±11)mm Hg to(22±7) mm Hg after shunt.There was no mortality perioperatively and during the follow-up.There were no recurrent hemorrhage nor hepatic encephalopathy occurring in the follow-up and all the children have normal intelligence and normal liver function though blood ammonia level increased significantly from(18±7)μmol/L to (60±17)μmol/L in 4 cases.In 7 cases in which preoperative whole blood cell count significantly decreased,the postoperative WBC,RBC,Hb and PLT was (7.64 ±4.46)×10~9/L,(4.54±0.97)×10~(12)/L,(133±5) g/L and (355.40±107.36)×10~9/L respectively (all P ＜0.05).In one case suffering from preoperative low PLT count the postop PLT reached 333×10~9/L,which was significantly higher than that preoperatively.Esophageal varices ameliorated in 6 cases.No stenosis of anastomotic stoma and thrombosis developed.Conclusion Spleno-left adrenal vein shunt is an effective procedure to treat portal vein cavernous transformation induced portal hypertension in children.

To study the variation and significance of plasma coagulation factor VII (FVII) in different kinds of ischemia heart disease (IHD) and examine its relation with plasma lipid and gene polymorphism. FVIIa was determined with one stage clotting assay by using a recombinant soluble tissue factor (rsTF). FVIIc was measured with one stage clotting assay. FVIIag was quantified with an enzyme-linked immunosorbent assay (ELISA). Polymorphism was analyzed with PCR-urea-polyacrylamide gel electrophoresis. Our results showed that FVIIa in stable angina (SA), unstable angina (UA), obsolete and acute myocardial infraction (OMI, AMI) patients was higher than those of normal group with the differences being significant within any two groups. FVIIag in UA, OMI and AMI was higher than those in SA and normal groups. There were positive correlations between FVIIa and serum triglycerides, FVIIa and FVIIc, FVIIc and FVIIag. FVII-323 0/10 bp polymorphism analysis was performed in 60 patients and 0/10 bp polymorphism was found in 5 cases. FVIIc and FVIIag were much lower in cases of 0/10 bp groups than those in cases of 0/0 bp groups. It is concluded that there was activation of extrinsic coagulation pathway in every kind of IHD to different extent. FVIIa was the risk factor in the development of IHD, and more sensitive in reflecting the severity of cardiovacutar disease than FVIIc or FVIIag. FVIIa was higher in OMI, which may be one of the risk factors of re-infraction. Serum triglyceride may indirectly lead to the development of IHD by increasing the level of FVIIa. FVII-323 0/10 bp polymorphism was present in Chinese patients with IHD and it was correlated with the level of FVIIc, FVIIag in plasma. 10 bp allelomorphic gene was a protective factor against thrombogenesis.

Objective To study the methodology and effect of auricle reconstruction with prefabricated expanded fascial flap in temporal and parietal area and skin flap in mastoid area, covered on the Medpor support. Methods Thirty cases of congenital vestigial auricle were involved in this study. The operations were divided into two stages: in the stage I,the temporal and parietal superficial fascia flap was cut with a vascular pedicle in the parietal and temporal superficial area, which was implanted under the skin of mastoid, and then both layers of those were expanded at the same time; in the stage II, this fascial skin flap was cut down and covered on the whole Medpor support to rebuild the man-made auricle. Results Thirty cases were successful after following-up for 6 months to 3 years. Their auricle shapes were vivid; their skin color was normal, and it had no any difference from all around skins, or no any appearance of the support. Conclusions With best understanding the biological characteristics of the Medpor support and choosing the suitable procedures and using the prefabricated expanded support, we could minimize the surgical complications in this operation. The results show that this procedure is satisfactory, without any necrosis of flap, swelling of contour, or appearance of the support. The combination of two flaps is the best method for the auricle reconstruction.

Objective:To explore the efficacy and safety of transurethral anatomical vapor incision technique of prostate (VIT) with six-step method high power side-emitting greenlight laser in the treatment of benign prostatic hyperplasia (BPH).Methods:A retrospective analysis of 82 patients with BPH who used high power side-out green laser in the treatment from October 2018 to June 2020 in Gongli Hospital of Naval Medical University was performed. Among them, 40 patients were treated with six-step method VIT, and 42 patients were treated with photoselective vaporization of prostate (PVP). The two groups of patients were compared in age [(71.1±8.7)years vs.(72.1±7.0)years], prostate volume [75 (68.25, 89.00) ml vs. 73 (63.25, 85.00) ml], and peak urinary flow rate (Q max) [6.20 (5.20, 8.20) ) ml/s vs. 5.9 (4.75, 7.50) ml/s], post-void residual volume (PVR) [74.00 (42.50, 103.75) ml vs. 67.00 (58.00, 84.50) ml], international prostate symptom score (IPSS) [(21.2±5.2) vs. ( 21.0±3.9)], quality of life score (QOL) [5 (4, 6) vs. 5 (4, 6) ], prostate specific antigen (PSA) [6.20 (4.12, 8.43) ng/ml vs. 5.40 (3.88, 7.13) ng/ml ]. In general, there was no statistical difference ( P>0.05). The VIT group adopts the six-step method of marking, removing film, grooving, excision, trimming and crushing. In the PVP group, the prostate tissue was uniformly vaporized layer by layer from the inside to the outside. Perioperative indexes and complications were compared between the two groups. The Q max, IPSS, QOL, PVR and PSA between the two groups before and 3 months after surgery were compared. Results:All patients in the VIT group and PVP group successfully completed the surgery, and there was no case of transfer to TURP or open surgery. The average operation time was [60.00(50.00, 73.75)min vs. 70.00(50.00, 73.75)min] ( P<0.05). There was no significant difference in the amount of postoperative hemoglobin decline[15.00(10.00, 17.75)g/L vs. 16.00(14.00, 19.25)g/L], average bladder irrigation time[1(1, 1)d vs. 1(1, 1)d], indwelling catheterization time[3(3, 3)]d vs. 3(3, 3)d] and hospitalization time in patients after operation[4(3, 4)d vs. 4(4, 4)d] ( P>0.05). All patients had no blood transfusion, second bleeding, readmission, TURS, urethral stricture and urinary incontinence.There were 2 cases (5.0%) of postoperative urinary tract infection in the VIT group and 9 cases (21.4%) of postoperative urinary tract infection in the PVP group ( P<0.05), and they were cured after anti-inflammatory treatment. Three months after operation, Q max, IPSS, QOL, PVR and PSA in the two groups were significantly improved compared with preoperatively. Among them, the differences of IPSS [(5.7±2.5) points vs. (7.5±2.8) points] and PSA [2.65(2.10, 3.90)ng/ml vs. 4.00(2.45, 4.45)ng/ml] in the VIT group and PVP group after operation were statistically significant ( P<0.05). Conclusions:Applying the six-step method high power side-emitting greenlight laser transurethral anatomical VIT to treat BPH, there is less intraoperative and postoperative bleeding, short operation time, significant decrease in PSA, and fewer complications. It is a safe and effective minimally invasive technology for the treatment of BPH.

OBJECTIVETo investigate the role of excretory/secretory antigens from Clonorchis sinensis (CsESAs) in hepatic fibrosis induced by C. sinensis infection in rats and explore the possible mechanism.

METHODSCsESAs was collected from adult C. sinensis cultured in sterile condition for 12 h and injected intraperitoneally in Wistar rats. Masson staining was used to observe the changes in the hepatic collagen fiber after the injection. HE staining and immunofluorescence staining were performed to detect the expression of alpha-smooth muscle actin (alpha-SMA) to examine the proliferation and the activity of hepatic stellate cells. The specific antibody titer of CsESAs was determined using enzyme-linked immunosorbent assay to investigate the role of the antigen-antibody complex in the development of hepatic fibrosis.

RESULTSAfter intraperitoneal injection of CsESAs, obvious hepatic fibrosis and hepatic stellate cell proliferation and activation were observed in the rat livers. The severity of the hepatic fibrosis was associated with the dose of CsESAs injected, whereas the titer of the specific antibody against CsESAs showed no direct relation to the hepatic fibrosis.

CONCLUSIONIntraperitoneal injection of CsESAs can cause hepatic stellate cell activation and hepatic fibrosis in rats, but the antigen-antibody complex does not seem to play the key role in the activation of the hepatic stellate cells.

Actins ; metabolism ; Animals ; Antigens, Helminth ; immunology ; Clonorchiasis ; parasitology ; Clonorchis sinensis ; immunology ; pathogenicity ; Hepatic Stellate Cells ; pathology ; Liver Cirrhosis ; immunology ; parasitology ; Male ; Rats ; Rats, Wistar

Objective:To study the role of Jagged1 and Notch signaling pathway played in the differentiation and proliferation of RAW 264.7 cells.Methods: RAW 264.7 cells were divided into three groups to culture:The control group:RAW 264.7 cells were threated with culture and RANKL.The Jagged1 group:RAW 264.7 cells were threated with recombinant protein Jagged 1 besides the control group.The DAPT group:RAW 264.7 cells were threated with DAPT besides the Jagged 1 group.The mRNA expression of osteoclast markers(TRAP,CK,CTR) and Notch key target genes (HES-1 and HEY-1) were measured by real-time PCR.The formation of osteoclast , bone resorption , Notch expression and proliferation of RAW 264.7 cells were detected by TRAP staining , scanning electron microscope ,immunofluorescence and cell counting kit-8 ( CCK-8 ).Results: TRAP, CK, CTR , HES-1 and HEY-1 mRNA expression were significantly higher than the control group and DAPT group in Jadded 1 group ( P<0.05 ).TRAP+cell count ,osteolytic area was significantly increased in Jagged 1 group compared with control and DAPT group , and no significant difference observed between the last two groups.Immunofluorescence results showed high expression of N ICD in cell membrane and cytoplasm in all groups and additionally expressed in nucleus in Jadded 1 group.Cell proliferation was inhibited in Jagged 1 group also ( P<0.05 ).Conclusion:Jagged1 promotes RAW264.7 cells osteoclast differentiation and inhibits proliferation by activating Notch signaling pathway .

Design space approach is applied in this study to enhance the robustness of first ethanol precipitation process of Codonopsis Radix (Dangshen) by optimizing parameters. Total flavonoid recovery, dry matter removal, and pigment removal were defined as the process critical quality attributes (CQAs). Plackett-Burman designed experiments were carried out to find the critical process parameters (CPPs). Dry matter content of concentrated extract (DMCE), mass ratio of ethanol to concentrated extract (E/C ratio) and concentration of ethanol (CEA) were identified as the CPPs. Box-Behnken designed experiments were performed to establish the quantitative models between CPPs and CQAs. Probability based design space was obtained and verified using Monte-Carlo simulation method. According to the verification results, the robustness of first ethanol precipitation process of Dangshen can be guaranteed by operating within the design space parameters. Recommended normal operation space are as follows: dry matter content of concentrated extract of 45.0% - 48.0%, E/C ratio of 2.48-2.80 g x g(-1), and the concentration of ethanol of 92.0% - 92.7%.
Chemical Precipitation ; Chemistry, Pharmaceutical ; methods ; Codonopsis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification

OBJECTIVETo clone the gene human thioredoxin 1 (hTrx-1) expressing its protein in the E.coli expression system and to obtain its polyclonal antibody, and to study the protective effects of hTrx-1 on neonatal rats with endotoxemia.

METHODSDNA encoding hTrx-1 from fetal liver cells was isolated by RT-PCR. The hTrx-1 was cloned to the prokaryotic expression plasmid PET-28a to induce its protein expression in the E.coli expression system. The purified hTrx-1 was injected into rats to prepare polyclonal antibody. Newborn Sprague-Dawley rats were randomly assigned to three groups: control, lipopolysaccharide (LPS) and hTrx-1 (n=12 each). The control and the LPS groups were intraperitoneally injected with normal saline and LPS (5 mg/kg), respectively. The hTrx-1 group received an intraperitoneal injection of hTrx-1 (10 mg/kg) 30 minutes before LPS injection. The mortality rate 24 hrs after injection was compared between the three groups.

RESULTSThe prokaryotic expression plasmid PET-28a-hTrx-1 was constructed. The hTrx-1 protein was expressed and purified. The polyclonal antibody of hTrx-1 with the titer of 1∶51200 was prepared. The mortality rate of the control, LPS and hTrx-1 groups was 0, 67% and 17%, respectively (χ2=14.400, P<0.01).

CONCLUSIONSThe polyclonal antibody of hTrx-1 is prepared successfully. The hTrx-1 protein has protective effects on neonatal rats with endotoxiamia.

Animals ; Animals, Newborn ; Antibodies ; analysis ; Blotting, Western ; Endotoxemia ; prevention & control ; Enzyme-Linked Immunosorbent Assay ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; immunology ; Thioredoxins ; genetics ; immunology ; therapeutic use

Objective To investigate antimicrobial resistance among gram-positive cocci in China in 2006.Methods From Jun 2006 to Dec 2006,674 consecutive and non-repetitive gram-positive bacteria were collected from 7 teaching hospitals.The minimal inhibitory concentration(MICs)of antibacterial agents were determined by agar dilution method.Results The prevalence of penicillin.resistant(ease)and pemcllhn. intermediate S.pneumoniae(PISP)among 100 isolates was l%and 19%,respectively.Teicoplanin and vancomycin were the most active agents against S.pneumoniae.97% and 98% S.pneumoniae isolates were susceptible to levofloxacin and moxifloxacin,respectively.The susceptibility of amoxicillin/clavulanic acid, ceftriaxone and chloramphenicol are 96%,87% and 73%,respectively.The susceptible rates of penicillin. susceptible S.pneumoniae(PSSP)to cefprozil and cefaclor were 62% and 55.7%,respectively.All the PISP and PRSP isolates were resistance to the two antibiotics.The susceptibility to macrolides,trimethoprim/ sulfamethoxazole and tetracycline was lower than 35%.The prevalence of methicillin-resistant Staphylococcus aureus(MRSA)and methicillin.resistant coagulase-negative Staphylococci(MRSCON)was 48%(33%-84%)and 81%(69%-94%),respectively.The susceptible rates of MRSA to trimethoprim/ sulfamethoxazole,chloramphenicol,rifampin,and the other antibiotics in this study were 72%,66%and 45%,respectively.The susceptible rate of MRSA to marcrolides,aminoglycosides,tetracyclines and quinolones were not more than 18%.56%(30%-86%)of E.faecalis and 80%(50%.100%)of E.faecium were highly resistant to gentamicin.The susceptibility of E.faecalis to all the antibiotics except chloramphenicol and tetracycline were higher than E.faecium.All isolates of S.aureus,CoNS and E.faecalis tested were susceptible to vacomycin and teicoplanin.There were two vacomycin.resistant E.faecium strains isolated from Hangzhou.Conclusions Antimicrobial resistance patterns of gram.positive cocci differed in different regions.The resistance of gram-positive cocci to the antibiotics in this study this year was a little higher than the data of the year of 2005.Teicoplanin and vancomycin remained very high activity to gram-positive cocci.