BACKGROUND:Vascular endothelial growth factor is the most important pro-angiogenic factor, which can promote revascularization and survival of fat grafts during fat transplantation. OBJECTIVE:To investigate the effect of vascular endothelial growth factor 165 transfection on the proliferation of adipose-derived mesenchymal stem cel s. METHODS:Recombinant vascular endothelial growth factor 165 mRNA fragment was transmitted into adenovirus pAdEasy-1 systems that were packaged to measure viral titer (experimental group). Empty adenovirus was also packaged as control group. Two kinds of packaged adenovirus solution were transferred into adipose-derived mesenchymal stem cel s at the best multiplicity of infection=100. Cel s with no transfection served as blank group. RT-PCR and western blot methods were used to detect the expression of vascular endothelial growth factor 165 at mRNA and protein levels;MTT method was adopted to detect cel proliferation. RESULTS AND CONCLUSION:The expressions of vascular endothelial growth factor 165 mRNA and protein were significantly higher in the experimental group than the control and blank groups (P<0.05). The division and proliferation of transfected adipose-derived mesenchymal stem cel s were increased significantly in the experimental group, which was significantly different from the control and blank groups (P<0.05). These finding indicate that vascular endothelial growth factor 165 transfection cannot only sustain the target protein expression of adipose-derived mesenchymal stem cel s, but also promote the proliferation of adipose-derived mesenchymal stem cel s remarkably.

BACKGROUND:Platelet-derived endothelial cel growth factor (PD-ECGF) can promote revascularization in fat transplantation. OBJECTIVE: To explore the dual effects of PD-ECGF and adipose-derived mesenchymal stem cels on the survival rate of fat grafts. METHODS:(1) Adipose-derived mesenchymal stem cels were isolated from the inguinal subcutaneous fat of New Zealand white rabbits, and then cultured. Passage 3 adipose-derived mesenchymal stem cels were divided into experimental group (Lenti-PD-ECGF-EGFP transfected adipose-derived mesenchymal stem cels), control group (Lenti-EGFP transfected adipose-derived mesenchymal stem cels) and blank group (adipose-derived mesenchymal stem cels with no transfection). (2) Lenti-PD-ECGF-EGFP transfected adipose-derived mesenchymal stem cels were cultured in DMEM complete medium, and then mixed with fat tissues as group A; adipose-derived mesenchymal stem cels with no transfection were cultured in DMEM complete medium and then mixed with fat tissues as group B; DMEM complete medium with no cels served as group C. Then, the grafts in groups A, B, C were respectively injected subcutaneously into the upper left, lower left and upper right parts of the rabbits’ black. RESULTS AND CONCLUSION:(1) In the experimental group, PD-ECGF mRNA and protein expressions were significantly higher than those in the control and blank groups (P < 0.05), and cel proliferation was also the fastest. (2) Graft weight and the number of capilaries were greater in group A than groups B and C. These findings indicate that PD-ECGF transfection of adipose-derived mesenchymal stem cels not only can continuously express the PD-ECGF protein, but also can promote the proliferation of adipose-derived mesenchymal stem cels.

OBJECTIVETo observe the curative effect of integrative medicine for treatment of systematic lupu erythematosus (SLE).

METHODSTotally 110 cases of SLE were randomized into two groups, 50 in the control group and 60 in the treated group, both were treated with prednisone, but to the treated group, integrative medica treatment was given additionally according to syndrome differentiation. The course for both groups was 6 months. Clinical symptoms, immunological indexes, peripheral blood figure, erythrocyte sedimentation rate (ESR), levels of C-reactive protein (CRP) and creatinine (Cr) in blood, and 24 h urinary total protein (u-TP/24 h) were observed before and after treatment.

RESULTSOf the 60 patients in the treated group, the treatment on 29 was evaluated as clinical remission, 18 as remarkably effective, 9 as effective, and 4 as ineffective, the total effective rate being 93.33% (56/60), while that in the control group was clinical remission in 11, remarkably effective in 10, effective in 19, ineffective in 10 respectively, and the total effective rate 80% (40/50), the difference on total ef fective rate between the two groups was significant (chi2 = 4.36, P <0.05). Besides, the improvement in the treated group was superior to that in the control group in terms of clinical symptoms such as fever, arthralgia and baldness (P <0.05, P <0.01); the negative reversion rate of anti-nuclear antibody (ANA, 51.3% vs 28.1%), antidouble-stranded DNA (ds-DNA, 53.6% vs 26.1%), anti- ribonucleoprotein (RNP, 63.2% vs 29.4%) and circulating immnue complexes (CIC, 63.2% vs 29.4%, all P <0.05); lowering of immunoglobulin (for IgG, P < 0.05; IgA, P <0.01; IgM, P <0.01); as well as bettering in peripheral blood figure, ESR, CRP, Cr and u-TP/24 h (P <0.05 or P <0.01).

CONCLUSIONIntegrative medical treatment is obviously effective for SLE patients.

Adolescent ; Adult ; Autoantibodies ; blood ; Blood Sedimentation ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Integrative Medicine ; Lupus Erythematosus, Systemic ; blood ; drug therapy ; immunology ; Male ; Middle Aged ; Prednisone ; therapeutic use ; Young Adult

OBJECTIVETo investigate the effects and mechanisms of Wuling mycelia on seizure development and learning ability induced by pentylenetetrazole-kindling epilepsy in rats.

METHODSSD rats were randomly divided into four groups: pentylenetetrazole-kindling model group (model group), low dose Wuling mycelia (0.3 g*kg(-1)) group (LD-WM group), high dose Wuling mycelia (0.6 g*kg(-1)) group (HD-WM group) and control group. The rats were intraperitoneal injected with a subconvulsive dose (35 mg*kg(-1)) of pentylenetetrazole (saline in control group) every 48 h for 12 times. Wuling mycelia was intragastrically applied 30 min before pentylenetetrazole injection. An 8-arm radial maze ( 4 arms baited) was used to measure the learning ability. Histamine was measured by chemical fluorometric enzyme immunoassay.

RESULTSCompared with the model group, the kindling stage of LD-WM group degraded significantly after 7th injection, the latency to the onset of myoclonic jerks (LTMJ) and the latency to the onset of generalized seizures (LTGS) prolonged after the 6th and 7th injection, respectively (P<0.05). The kindling stage of HD-WM group also degraded markedly after the 6th to 8th injection, and the LTMJ and the LTGS extended after the 8th to 9th and 6th injection, respectively (P<0.05). Compared with the control group, the frequency of working memory error (WME) and reference memory error (RME) of the model group in the 8-arm radial maze increased through 3-d training (P<0.05). The memory tests showed that the impairment induced by pentylenetetrazole was partially reversed by Wuling mycelia. Compared with the control group, brain histamine contents (hippocampus, cortex, thalamus and hypothalamus) were significantly lower in model group (P<0.05). But compared with the model group, hippocampal histamine contents in LD-WM group and hippocampal, thalamic and hypothalamic histamine contents in HD-WM group were elevated (P<0.05).

CONCLUSIONWuling mycelia can delay the kindling and ameliorate the ability of learning in rats with pentylenetetrazole-induced epilepsy and the enhancement of neuronal histamine activity may be one of possible mechanisms.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Epilepsy ; chemically induced ; metabolism ; prevention & control ; Hippocampus ; metabolism ; Histamine ; metabolism ; Kindling, Neurologic ; drug effects ; Learning ; drug effects ; Male ; Pentylenetetrazole ; toxicity ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the influence of temperature on the expressions of c-kit and PI3K in spermatogonia cultured in vitro at 32 degrees C and 37 degrees C, and provide basic scientific data for the mechanism of spermatogenic impairment due to body temperature (37 degrees C).

METHODSIsolated spermatogenic cells were cultured in vitro at 32 degrees C and 37 degrees C, and their adherence, proliferation and morphologic changes were observed and recorded under the inverted phase contrast microscope. At 8 days, the spermatogonia were separated by Percoll density gradient centrifugation and the differential adhesion method. The expressions of c-kit and PI3K mRNA and proteins in the separated cells were detected by real time polymerase chain reaction and Western blot, respectively. The c-kit gene was sequenced to identify the occurrence of mutations.

RESULTSAdherence, division and proliferation of the cells were observed in both the 32 degrees C and 37 degrees C groups. The expressions of c-kit and PI3K mRNA and proteins in the spermatogonia were significantly higher in the 32 degrees C group than in the 37 degrees C group (P < 0.05). The 32 degrees C group showed no mutation of c-kit in exon 9, 11 and 13; the 37 degrees C group exhibited no mutation in exon 11 and 13, but possible insertion or deletion mutations in exon 9.

CONCLUSIONCulturing in vitro at 37 degrees C could inhibit the expression of proliferation- and differentiation-related genes in spermatogenic cells and lead to the mutation of the c-kit gene.

Base Sequence ; Cells, Cultured ; Exons ; Humans ; Male ; Mutation ; Phosphatidylinositol 3-Kinase ; genetics ; Proto-Oncogene Proteins c-kit ; genetics ; Spermatogenesis ; genetics ; Spermatogonia ; cytology ; Temperature

BACKGROUNDLaparoscopic splenectomy (LS) has been considered as the standard approach to remove a normal-sized spleen, but it is facing technical challenges when applied to splenomegaly. Hand-assisted laparoscopic technique was designed to facilitate the performance of difficult laparoscopic procedure. This study was aimed to evaluate the efficacy and superiority of hand-assisted laparoscopic splenectomy (HALS) for splenomegaly.

METHODSFrom November 1994 to January 2006, 36 patients with splenomegaly (final spleen weight > 700 g) were treated with laparoscopic operations for splenectomy in our hospital. Conventional LS was performed in 16 patients (7 men and 9 women, group 1) and HALS in the other 20 patients (12 men and 8 women, group 2). The patients' features, intraoperative details and the postoperative outcomes in the both groups were compared.

RESULTSThe both groups were comparable in the terms of patient's age ((38 +/- 12) years vs (43 +/- 14)years, P > 0.05), the greatest splenic diameter ((24 +/- 5)cm vs (27 +/- 7)cm, P > 0.05), preoperative platelet count ((118 +/- 94) x 10(9)/L vs (97 +/- 81) x 10(9)/L, P > 0.05) and diagnosis. Compared with LS group, operation time ((195 +/- 71) minutes vs (141 +/- 64) minutes, P < 0.05) was shorter, intraoperative blood loss ((138 +/- 80)ml vs (86 +/- 45)ml, P < 0.05) and conversion rate (4/16 vs 0/20, P < 0.05) were lower, but hospital stay ((5.3 +/- 3.8) days vs (7.4 +/- 1.6) days, P < 0.05) was longer in HALS group. There was no significant difference in the aspects of intraoperative and postoperative complication rate (2/16 vs 0/20, P > 0.05) or recovery time of gastrointestinal function ((16.3 +/- 11.6) hours vs (18.7 +/- 8.1) hours, P > 0.05) between the two groups.

CONCLUSIONSIn the cases of splenomegaly, HALS significantly facilitates the surgical procedure and reduces the operational risk, while maintaining the advantages of conventional LS. HALS is more feasible and more effective than conventional LS for the removal of splenomegaly.

Adolescent ; Adult ; Aged ; Female ; Humans ; Laparoscopy ; methods ; Male ; Middle Aged ; Splenectomy ; methods ; Splenomegaly ; surgery

OBJECTIVETo discuss the role of gadolinium chloride (GdCl(3)) in lung injury associated with acute necrotizing pancreatitis (ANP).

METHODSExperimental animals were randomized into five groups (n = 18 for each group): normal control group, ANP group, GdCl(3) pretreatment group, ANP GdCl(3) pretreatment group, ANP GdCl(3) treatment group. Rat ANP model was induced by intraductal administration of 3% sodium taurocholate. Alveolar macrophages (AM) were obtained by bronchoalveolar lavage. The blood gas assay, the ratio of wet/dry tissue, protein content of bronchoalveolar lavage fluids (BALF), the myeloperoxidase (MPO) of lung tissue and generation of TNFalpha and NO by AM were evaluated. The apoptosis of AM was checked by agarose gel electrophoresis analysis, transmission electric microscopy observation and cytometry propidium iodide single stained method. The lung tissue was examined by histology.

RESULTSThe parameter of GdCl(3) pretreatment group compared with normal control group had no statistical significance (P > 0.05). The indicators of ANP GdCl(3) pretreatment group and ANP GdCl(3) treatment group were elevated compared with the normal control group and had statistical significance (P < 0.05). But compared to the ANP group, they were all decreased and also had the statistical significance (P < 0.05). The 180 - 200 bp ladder pattern unique to apoptosis in agarose gel electrophoresis and the apoptotic typical morphologic feature in AM by transmission electric microscopy and typical subdiploid peak in DNA content figure could be observed in ANP GdCl(3) pretreatment group and ANP GdCl(3) treatment group, while the other three groups could not.

CONCLUSIONSLung injury associated with ANP could be ameliorated by application of GdCl(3) through inducing apoptosis of AM of ANP.

Animals ; Apoptosis ; drug effects ; Female ; Gadolinium ; therapeutic use ; Macrophages, Alveolar ; cytology ; drug effects ; Male ; Pancreatitis, Acute Necrotizing ; complications ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Respiratory Distress Syndrome, Adult ; etiology ; pathology ; prevention & control

OBJECTIVETo investigate the effects of nitric oxide (NO) on reperfusion injury following pancreaticoduodenal transplantation in rats.

METHODSThe homologous male Wistar rat model of heterotopic total pancreaticoduodenal transplantation was used. The L-arginine (L-Arg) group received intravenous injection of L-Arg 5 minutes before and after reperfusion at a dose of 200 mg/kg while the N-Nitro-L-Arginine methyl ester (L-NAME) group received intravenous injection of L-NAME at a dose of 10 mg/kg, and control group received saline. The amount of NO in the pancreas graft was measured. Serum concentration of cytokine-induced neutrophil chemoattractant (CINC) determined by enzyme-linked immunosorbant assay, expression of CINC mRNA detected by Northern blot assay, and myeloperoxidase (MPO) activity in the pancreas graft were measured. Histological observation was performed.

RESULTSThe amount of NO in the L-Arg group was higher than in the control group, while in the L-NAME group was lower than in the control group (P < 0.05). The peak of serum CINC concentration occurred 3 hours after reperfusion with significant difference among groups. Expression peak of CINC mRNA in the pancreas graft occurred 3 hours after reperfusion. The expression level in the L-Arg group was lower than in the control group, the L-NAME group was higher than control group (P < 0.05). MPO activity in the L-Arg group obviously decreasd compared with other groups. The pancreas inflammation was ameliorated in L-Arg group, and pancreas damage was aggravated in L-NAME group.

CONCLUSIONSL-Arg can increase the amount of NO and inhibit the elevation of CINC, CINC mRNA expression, and early neutrophil accumulation in the transplanted pancreas. NO has protective effects on the ischemia/reperfusion injury of pancreaticoduodenal transplantation.

Animals ; Arginine ; pharmacology ; Chemokines, CXC ; biosynthesis ; genetics ; Duodenum ; transplantation ; Male ; NG-Nitroarginine Methyl Ester ; pharmacology ; Nitric Oxide ; metabolism ; Pancreas Transplantation ; Peroxidase ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; Reperfusion Injury ; metabolism

OBJECTIVETo study mechanismt of Fufang Haishe capsule for dementia by observing the effect of it on PC-12 cell apoptosis, which was induced by beta-amyloid protein (Abl-42).

METHODNerve growth factor (NGF) was used to cultivate the PC-12 cells. Fufang Haishe capsule at different concentrations was added into the culture medium so as to identify the nontoxic concentrations with MTT. To analyze the PC-12 cell apoptosis respectively by MTT assay, Flow cytometry (FCM technique) with different concentrations of Fufang Haishe capsule (0.01, 0.1, 1, 5 mg x mL(-1)), adding Ab or not Western blot was used to detect apoptosis which was measured on the implementation of caspase-9 and caspase-3 activity.

RESULTFufang Haishe capsule could significantly inhibit the apoptosis of PC-12 cells induced by Abeta with increased colorimetric MTT asay ( compare among the control group and concentration 0, 0.01, 0.1, 1 and 5 mg x mL(-1) group, which is the same below: 1.75 +/- 0.12, 0.73 +/- 0.35, 0.79 +/- 0.11, 0.83 +/- 0.07, 1.31 +/- 0.07, 1.80 +/- 0.38, P < 0.01) and the decreased apoptosis rate of the cells which was analysed by flow cytometry (1.93 +/- 0.41)%, (46.17 +/- 4.08)%, (35.35 +/- 4.63)%, (28.62 +/- 3.81)%, (15.13 +/- 3.15)%, (7.84 +/- 1.76)%, P < 0.01. In addition, Fufang Haishe capsule inhibited the activity of caspase-9 and caspase-3 of PC-12 cells which was induced by Abeta.

CONCLUSIONFufang Haishe capsule significantly inhibite apoptosis of PC-12 cells induced by Abeta. The mechanism might be that Fufang Haishe capsule decrease the activity of the apoptosis implementing protein,caspase-9 and caspase-3.

Animals ; Apoptosis ; drug effects ; Capsules ; Caspases ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; PC12 Cells ; Rats

BACKGROUNDPrimary liver cancer (PLC) is a common malignant tumor. Over the past decade, although farnesyltransferase (FTase) has emerged as a significant target for anticancer therapies and has become a hotspot of cancer research, its exact mechanism of action remains unknown. The aim of this study was to investigate the expression of FTase in PLC and its role in the development of PLC.

METHODSExpression of FTase was detected by real-time fluorescent quantitative-polymerase chain reaction (FQ-PCR) in cancer and surrounding normal tissues from 32 patients with PLC.

RESULTSExpression of FTase mRNA in PLC was significantly higher than that in normal hepatic tissues (P < 0.001). Overexpression of FTase was as high as 87.5%. The positive rate for FTase mRNA in the high tendency to metastatic recurrence group was obviously higher than that in the low tendency to metastatic recurrence group (P = 0.02). The positive rate for FTase mRNA in patients with metastatic recurrence during postoperative follow-up was also significantly higher than that in those without metastatic recurrence (P = 0.01).

CONCLUSIONSThe level of FTase mRNA expression in cancer tissues is much higher than in normal tissues. FTase may play an important role in the genesis and development of PLC and may be one of the reliable markers for the metastatic activity gained by liver tumor cells. FTase could be used clinically in predicting metastatic recurrence of PLC.

Adult ; Aged ; Farnesyltranstransferase ; genetics ; Female ; Humans ; In Vitro Techniques ; Liver Neoplasms ; enzymology ; pathology ; Male ; Middle Aged ; Polymerase Chain Reaction ; RNA, Messenger ; Young Adult