Pirfenidone alleviates urethral stricture following urethral injury in rats by suppressing TGF-β1 signaling and inflammatory response / 南方医科大学学报

OBJECTIVE@#To investigate the effect of pirfenidone for reducing urethral stricture following urethral injury in rats and explore the possible mechanism.@*METHODS@#Thirty male SD rats were randomly assigned into negative control group, positive control group and pirfenidone group (n=10). In pirfenidone and positive control groups, the rats were subjected to incision of the posterior urethral cavernous body followed by daily intraperitoneal injection of pirfenidone (100 mg/kg) and an equivalent volume of solvent, respectively. The rats in the negative control group were given intraperitoneal injections of solvent without urethral injury. At two weeks after modeling, retrograde urethrography was performed for observing urethral stricture, and the injured urethral tissues were harvested for HE staining, Masson staining, immunohistochemical staining and Western blotting for detecting the protein expressions of α-SMA and TGF-β1. The mRNA expressions of the inflammatory factors TNF-α, IL-6, and IL-1β were detected using qRT-PCR.@*RESULTS@#The body weight of the rats in pirfenidone group was significantly decreased compared with that in the other two groups (P < 0.05). Retrograde urethrography showed significant narrowing of the urethra in the positive control group but not in the pirfenidone group. HE staining of the injured urethral tissues showed obvious proliferation of urethral epithelial cells with narrow urethral cavity and increased inflammatory cells in positive control group. The pathological findings of the urethra were similar between pirfenidone group and the negative control group. Masson staining revealed obviously reduced collagen fibers and regular arrangement of the fibers in pirfenidone group as compared to the positive control group. Compared with those in the negative control group, the expressions of α-SMA and TGF-β1 were significantly increased in the positive control group, and pirfenidone treatment significantly inhibited their expressions (P < 0.05 or 0.01). Pirfenidone also significantly inhibited the mRNA expressions of TNF-α, IL-6, and IL-1β in the injured urethral tissue (P < 0.05 or 0.01).@*CONCLUSION@#Pirfenidone can prevent urethral fibrosis and stricture after urethral injury possibly by inhibiting the TGF-β1 pathway and inflammatory response.

A case of acute carbon monoxide poisoning with secondary intestinal obstruction and thrombosis / 中华劳动卫生职业病杂志

Acute carbon monoxide poisoning can cause multiple organ damage due to hypoxia. In severe cases, it can be life-threatening and has a high fatality rate. Intestinal obstruction and thrombosis are rare complications of carbon monoxide poisoning. A case of carbon monoxide poisoning was reported. In addition to the central nervous system lesion, intestinal obstruction and lower limb thrombosis were also found. In the treatment of carbon monoxide poisoning patients, the clinician was able to treat the common complications, attention should be paid to gastrointestinal tract, thrombotic disease and other rare complications, so as to avoid missed diagnosis.

Effective Prevention of Acute Hemolytic Transfusion Reaction again by Blood Matching of in vitro Hemolytic Test / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To screen out the suitable erythrocytes compatible to the results from the routine blood matching for the patients suffered from the relapse of acute hemolytic transfusion reaction.</p><p><b>METHODS</b>The in vitro hemolysis test was used to screen out the erythrocytes from the non-hemolytic donors for the transfusion of erythrocytes into the patients.</p><p><b>RESULTS</b>Three U of the non-hemolytic erythrocytes were obtained by using hemolytic test in vitro, the post-transfusion effects were good, and the hemolytic reaction will not occure once more.</p><p><b>CONCLUSION</b>When it is compatible to the results obtained from the routine blood matching and the post-transfusion hemolytic reaction appeared. The blood matching by using in vitro hemolytic test can be used to screen out the non-hemolytic erythrocytes for transfusion of the patients.</p>

Effect of Calcium-sensing Receptor on the Apoptosis of Rat Spinal Cord Neurons in Anoxia/Reoxygenation Injury and Its Significance / 中国医学科学院学报

Objective To investigate the effect and significance of calcium-sensing receptor (CaSR) on the apoptosis of rat spinal cord neurons in anoxia/reoxygenation(A/R) injury. Methods The spinal cells were in ischemia and hypoxia environment for 1 h and in normal environment for 24 h to establish a model of A/R. After spinal A/R model was established,the spinal cells were divided into four groups randomly:the control group,A/R group,A/R+GdCl3 group,and A/R+NPS-2390 group. The expression of CaSR in each group was detected by immunofluorescence and Western blotting. The concentration of intracellular calcium was measured by laser confocal scanning microscopy. The expressions of Caspase-3,Bax,and Bcl-2 were detected by using Western blotting. The apoptotic rate of spinal cells was detected by Tunel assay. Results Compared to the control group, there was a significant increase in the level of CaSR (t=5.462, P=0.006), the concentration of intracellular calcium (t=8.573, P=0.001), the apoptotic rate (t=4.899, P=0.008), Caspase-3 (t=5.118, P=0.007), and Bax (t=10.930,P=0.001) in A/R group. Compared to the A/R group, there was a significant increase in the level of CaSR (t=4.975, P=0.008),the concentration of intracellular calcium (t=4.899, P=0.008), the apoptotic rate (t=7.746, P=0.002), Caspase-3 (t=4.776, P=0.009), and Bax (t=5.281, P=0.006) in A/R+GdCl3 group. Compared to the A/R group, there was a significant decrease in the level of CaSR (t=3.674,P=0.021), the concentration of intracellular calcium (t=3.846, P=0.018), the apoptotic rate (t=4.281,P=0.013), Caspase-3 (t=3.521, P=0.024), and Bax(t=3.473, P=0.026) in A/R+NPS-2390 group. However, compared to the control group, there was a significant decrease in the level of Bcl-2 (t=6.242,P=0.003) in A/R group. Compared to the A/R group, there was a significant decrease in the level of Bcl-2(t=3.028, P=0.004) in A/R+GdCl3 group. Compared to the A/R group, there was a significant increase in the level of Bcl-2 (t=2.840, P=0.047) in A/R+NPS-2390 group.Conclusion During the process of A/R injury in rat spinal cord neurons,the expression of calcium sensing receptor increases,along with increase in intracellular calcium and spinal neuron apoptosis.

Effect of Basic Fibroblast Growth Factor and Transforming Growth Factor-Β1 Combined with Bone Marrow Mesenchymal Stem Cells on the Repair of Degenerated Intervertebral Discs in Rat Models / 中国医学科学院学报

<p><b>OBJECTIVE</b>To evaluate the effects of the combination of basic fibroblast growth factor (bFGF), transforming growth factor-Β1 (TGF-Β1), bone marrow mesenchymal stem cells (BMSCs), and temperature-responsive chitosan hydrogel (TCH) gel on the repair of degenerative intervertebral disc in rat models.</p><p><b>METHODS</b>Rat models of intervertebral disc degeneration were established by acupuncture. The degenerative effects were observed under magnetic resonance imaging (MRI). The BMSCs was cultured in vitro and then transfected by adenovirus with enhanced green fluorescent protein to make it carry the gene of enhanced green fluorescent protein,which functioned as fluorescence labeling. The SD rat models of intervertebral disc degeneration were divided into four groups: group A, treated with the combination of bFGF, TGF-Β1,BMSCs,and TCH gel; group B, treated with the combination of BMSCs and TCH gel;group C, treated with the combination of bFGF,TGF-Β1, and TCH gel;and group D, treated with PBS buffer solution. After the corresponding reagents were injected into the degenerative intervertebral discs of each group, the rats were cultivated for another four weeks and then the repair effects of the intervertebral discs were observed under MRI. Furthermore,the intervertebral discs of each group were taken out and observed by HE and Masson staining. The nucleus pulposus was aspirated and the expressions of aggrecan,collagen 2,Sox-9,and collagen I of nucleus pulposus of each group were tested by reverse transcription polymerase chain reaction and Western blot.</p><p><b>RESULTS</b>The transplanted BMSCs survived in the intervertebral disc and differentiated into nucleus pulposus-like cells. MRI showed that:the signal intensity of the nucleus pulposus of group A was much higher than that of the rest groups, the signal intensity of group B was higher than that of group C, and the signal intensity of group D was the lowest,in which the dura mater spinalis was in compression and the spinal cord changed in beaded shape. The differences of the Pfirrmann grading among the four groups had statistical significance (P<0.05). The results of the HE and Masson stains showed:the intervertebral disc of group A was well-structured,the quantity of nucleus pulposus cells was larger than that of the other three groups,and the boundary between the nucleus pulposus and the annulus fibrosus was clearly defined;the quantity of the nucleus pulposus cells of group B was larger than that of group C, and the broken annulus fibrosus was not observed in group B, while the broken annulus fibrosus could be observed in group C; and, the nucleus pulposus cells of group D were replaced by fibrous tissue. The results of the reverse transcription polymerase chain reaction and Western blot tests showed that,in terms of the expressions of aggrecan,collagen 2 and Sox-9,group A was the highest, followed by group B,group C,and group D (P<0.05); in terms of the expression of collagen 1,there was no obvious difference among these four groups (P>0.05).</p><p><b>CONCLUSIONS</b>The transplanted BMSCs can survive in the degenerative intervertebral disc and differentiate into nucleus pulposus-like cells. The combination of bFGF, TGF-Β1, BMSCs,and TCH gel has obvious repair effect on the degenerative intervertebral discs. The effect of the combination of BMSCs and TCH gel on transplantation therapy of the degenerative intervertebral discs is better than that of the combination of bFGF, TGF-Β1 and TCH gel but worse than that of the combination of bFGF, TGF-Β1, BMSCs, and TCH gel.</p>

Role of bone marrow mesenchymal stem cells in restoring the functions of degenerative nucleus pulposus cells / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore the effects of bone mesenchymal stem cells (BMSCs) in restoring the functions of degenerative nucleus pulposus cells (dNPCs).</p><p><b>METHODS</b>The animal models of degenerative nucleus pulposus were established by means of acupuncture and aspiration. The BMSCs as well as the normal and degeneratived nucleus pulposus cells of SD rats were isolated and cultured. The BMSCs/alginate gel complex and dNPCs/alginate gel complex were used for indirect co-culture in vitro, which was set as experiment group. The NPCs and dNPCs cultured alone as positive and negative controls. The cell growth conditions were observed by light microscopy, and suitable cells were selected to combine alginate gel stents and cultured in transwell plate. Seven days later, nucleus pulposus cells of each group were recycled, and the mRNA expressions of Collagen2, SOX 9, and Aggrecan were detected by RT-PCR, and the Collagen1, 2, and Aggrecan were detected by Western blotting and immunofluorescence.</p><p><b>RESULTS</b>After non-contact co-culture for 7 days, the mRNA levels of Collagen2, SOX 9, and Aggrecan increased apparently in BMSCs+dNPCs group, while it was significantly lower in dNPCs sample (all P<0.05). The content of Collagen2 and Aggrecan detected by Western blotting in BMSCs+dNPCs group got close to NPCs sample, but it was significantly higher than dNPCs samples (all P<0.05), while the content of Collagen1in BMSCs+dNPCs group got close to NPCs samples, but it was significantly lower than dNPCs sample (P<0.05). Immunofluorescence results showed that cytoplasm was dyed red and the color near the caryon became dark in BMSCs+dNPCs group and dNPCs group;however, the colored scope of cytoplasm and the dark colored scope surrounding the caryon in BMSCs+dNPCs group was obviously larger than dNPCs group.</p><p><b>CONCLUSION</b>Under a 3D non-contact co-culture system, BMSCs can promote the expression of epimatrix of the dNPCs, which shows that BMSCs can restore the functions of dNPCs of intervertebral disc to certain extent.</p>

Prospective experimental studies on the renal protective effect of ulinastatin after paraquat poisoning / 世界急诊医学杂志（英文）

BACKGROUND: Paraquat (PQ) is an effective herbicide and is widely used in agricultural production, but PQ poisoning is frequently seen in humans with the lung as the target organ. Currently, there are many studies on lung injury after PQ poisoning. But the kidney as the main excretory organ after PQ poisoning is rarely studied and the mechanisms of this poisoning is not very clear. In this study, we observed the expression of caspase-3 and livin protein in rat renal tissue after PQ poisoning as well as the therapeutic effects of ulinastatin. METHODS: Fifty-four Sprague-Dawley (SD) rats were randomly divided into three experimental groups: control group (group A), paraquat poisoning group (group B) and ulinastatin group (group C), with 18 rats in each group. Rats in group B and group C were administered intragastrically with 80 mg/kg PQ, rats in group C were injected peritoneally with 100000 U/kg ulinastatin once a day, while rats in group A were administered intragastrically with the same volume of saline as PQ. At 24, 48, 72 hours after poisoning, the expression of livin in renal tissue was detected by Westen blotting, the expression of caspase-3 was detected by immunohistochemistry, and the rate of renal cell apoptosis was tested by TUNEL detection. The histopathological changes were observed at the same time. RESULTS: Compared to group A, the expression of caspase-3 in the renal tissue of rats in groups B and C increased significantly at any time point. Compared with group B, the expression of caspase-3 in renal tissue of rats in group C decreased. Compared with group A, the expression of livin in renal tissue in rats of groups B and C increased significantly at any time point (P<0.01), especially in group C (P<0.01). TUNEL method showed that the rate of renal cell apoptosis index was higher in group B at corresponding time points than in group A (P<0.01), and was lower in group C at corresponding time points than in group B (P<0.01). CONCLUSION: UTI has a protective effect on the renal tissue of rats after paraquat poisoning through up-regulating the expression of livin and down-regulating the expression of caspase-3, but the regulation path still needs a further research.

Effect of ulinastatin on the expression of heat shock protein 70 and NF-kappaB in lung tissue in rats with paraquat poisoned / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To observe the expression levels of heat shock protein 70 (hsp70) and NF-kappaB p65 mRNA in lung tissue of acute paraquat (PQ) poisoning rats, and intervention effects of ulinastatin (UTI).</p><p><b>METHODS</b>Seventy-two Sprague-Dawley (SD) rats were randomly divided into three groups: PQ poisoning group, UTI group and control group. The rats were exposed intragastrically to PQ at the dose of 80 mg/kg to establish a model of the rat acute lung injury. The UTI group was intervened by peritoneal injection with 10000 U/kg UTI in 30 minutes. On the 12, 24, 48, 72 h after exposure, myeloperoxidase (MPO) activity in lung tissue were detected. The expression of the NF-kappaB p65 mRNA and hsp70 mRNA in lung tissue was detected by the reverse transcription-PCR (RT-PCR). The lung pathological changes of rats were observed.</p><p><b>RESULTS</b>The degree of lung injury in PQ group and UTI group was higher than that in control group. But in UTI group the degree of lung injury was lower than PQ group. MPO activity in the lung tissues in PQ group was (31.72 +/- 6.42), (56.23 +/- 8.63), (87.21 +/- 10.02) and (107.21 +/- 13.52) micro/g in 12, 24, 48 and 72 h, respectively which was significantly higher than that [(11.38 +/- 1.25) micro/g] in control group (P < 0.01). MPO activity in the lung tissues in UTI group was (15.65 +/- 3.21), (35.98 +/- 5.74), (59.33 +/- 9.65) and (71.25 +/- 10.58) micro/g in 12, 24, 48 and 72 h, respectively which was significantly lower than those in PQ group (P < 0.01). The expression levels of NF-kappaB p65 mRNA of lung tissues in UTI group in 12, 24, 48 and 72 h were 0.3288 +/- 0.0147, 0.5337 +/- 0.0328, 0.7357 +/- 0.0424 and 0.7547 +/- 0.0905, respectively, which were significantly lower that those (0.4185 +/- 0.0294, 0.8532 +/- 0.0841, 0.9554 +/- 0.0975 and 1.0094 +/- 0.0703) in PQ group (P < 0.01). hsp70 mRNA expression levels in 12, 24, 48 and 72 h of the UTI group were 0.5193 +/- 0.0254, 0.8289 +/- 0.0606, 0.7566 +/- 0.0277 and 0.4873 +/- 0.0105, respectively, which were significantly higher than those (0.3897 +/- 0.0125, 0.5904 +/- 0.0186, 0.4007 +/- 0.0237 and 0.2293 +/- 0.0137) in PQ group (P < 0.01).</p><p><b>CONCLUSION</b>The expression levels of hsp70 mRNA and NF-kappaB p65 mRNA of rats after intoxication increased significantly. UTI can protect the lung tissues by elevating the expression of hsp70 and reducing the expression of NF-kappaB in the lung tissues of rats with acute paraquat poisoning.</p>

Nerve growth factor expression in astrocytoma and cerebrospinal fluid: a new biomarker for prognosis of astrocytoma / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Recent studies have discovered that nuclear translocation of nerve growth factor (NGF) and its receptor fragments function differently from the traditional model. This study aimed to uncover the nuclear expression of NGF in astrocytoma and its biological significance.</p><p><b>METHODS</b>Ninety-four paraffin-embedded astrocytoma specimens were subjected to immunohistochemical (IHC) and hemotoxylin & eosin (HE) staining. Preoperative cerebrospinal fluid (CSF) specimens and intraoperative snap-frozen astrocytoma tissues were assayed for NGF expression by ELISA and Western blotting. The outcome of patients who contributed samples was tracked. Each ten tissue samples from patients with traumatic brain injury who had received decompression surgery and CSF samples from patients undergoing spinal anesthesia but with no history of nervous system disease were taken as control.</p><p><b>RESULTS</b>NGF-positive immunoreactive products were distributed in both the cytoplasm and nucleus of astrocytoma, but were only located in the cytoplasm of traumatic brain injury (TBI) tissue. NGF nuclear-positive rate (NPR) of grades III - IV astrocytomas (70.0%) was higher than that of grades I - II astrocytoma (28.6%, P < 0.05). NGF-NP expression positively correlated with the NGF concentration in cerebrospinal fluid (CSF) (r = 0.755, P < 0.01). Kaplan-Meier survival analysis indicated that the median survival time was 25 months for NGF-NP astrocytoma grade I - II patients and 42 months in NGF nuclear negative (NGF-NN) astrocytoma grade I - II patients (P < 0.05). In astrocytoma III - IV patients, the median survival was 7 months for NGF-NP patients and 24 months for NGF-NN patients (P < 0.01). Two types of NGF with molecular weights of 13 and 36 kDa were present in astrocytoma, but only the 36 kDa NGF was found in the CSF. NGF expression elevated as the malignancy increased.</p><p><b>CONCLUSIONS</b>NGF-NP expression and NGF level in CSF were significant prognostic factors in astrocytoma patients. Because of the easy access of CSF, it may be developed as an index for early diagnosis and surveillance of astrocytoma.</p>

Expression of heat shock protein 70 in lung tissues of acute paraquat poisoned rats and intervention of ulinastatin / 世界急诊医学杂志（英文）

@#BACKGROUND: Paraquat (PQ) is an effective herbicide and is widely used in agricultural production, but PQ poisoning is frequently seen in humans with the lung as the target organ. Clinically pulmonary pathological changes are often used to predict the severity and prognosis of the patients. In this study, we observed the expression of heat shock protein 70 (HSP70) in rat lung after PQ poisoning and to investigate the therapeutic effects of ulinastatin. METHODS: Seventy-two adult healthy SD rats were randomly divided into a control group (group A, n=24), a poisoning group (group B, n=24), and an ulinastatin group (group C, n=24). The rat models of acute PQ poisoning were established by intra-gastric administration of 80 mg/kg PQ to rats of groups B and C, and the rats of group C were intra-peritoneally injected with 100000 IU/kg ulinastatin 30 minutes after poisoning. The expression of HSP70 in lung tissue was observed, and W/D and histopathological changes in the lung tissue were compared 12, 24, 48 and 72 hours after poisoning. The expression of HSP70 in the lung tissue was assayed by using RT-PCR. All quantitative data were processed with one-way analysis of variance to compare multiple sample means. RESULTS: Compared to group A, the expression of HSP70 in the lung of rats in groups B and C increased significantly at all intervals (P<0.05). The pathological changes in lung tissue of rats with PQ poisoning included congestion, leukocytes infiltration and local hemorrhage, whereas those of group C were significantly lessened. CONCLUSION: Ulinastatin may ameliorate acute lung injury to some extent after PQ poisoning in rats by enhancing the expression of HSP70.

A novel biosynthetic hybrid scaffold seeded with olfactory ensheathing cells for treatment of spinal cord injuries / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Implantation of tissue-engineered scaffolds is one of the most promising therapeutic strategies for inducing nerve regenerations following spinal cord injuries. In this paper, we report a novel bioengineered hybrid scaffold comprised of three major extracellular matrix (ECM) proteins.</p><p><b>METHODS</b>ECM-scaffolds (ECM-S) were prepared by gelling fibrinogen, fibronectin and laminin using fresh rat plasma. Olfactory ensheathing cells (OECs) were isolated from fresh rat olfactory mucosa, purified under differential adhesion, and assessed by immunofluorescent staining. OECs were seeded onto ECM-S and cultured. The effects of the scaffolds on the seeded cells were detected using the immunofluorescent staining, Western blotting, scanning electron microscopy and transmission electron microscopy.</p><p><b>RESULTS</b>Tissue-engineered ECM-S could be easily molded into mat-like or cylindrical shapes and gelled by addition of fresh plasma. Observations by electron microscopy show that the ECM-S forms a stable three-dimensional porous network. Studies on the effects of the ECM-S on the biological behaviors of OECs in vitro indicate that the scaffold can promote OEC adhesion, proliferation and process extensions. Additionally, OECs seeded on the scaffold maintained the expression of nerve growth factor, matrix metalloproteinase-3 and matrix metalloproteinase-9.</p><p><b>CONCLUSION</b>We developed a biosynthetic hybrid gel which could be used as a scaffold for OEC transplantation; this gel can promote nerve regeneration following spinal cord injuries.</p>

The efficacy of biological dressing containing calcium and magnesium on the management of hydrofluoric acid burns / 中华烧伤杂志

<p><b>OBJECTIVE</b>To observe the efficacy of biological dressing containing calcium and magnesia (sheep dermis absorbing calcium and magnesia and cross-link with glutaraldehyde) on the management of hydrofluoric acid burns in rats and patients.</p><p><b>METHODS</b>Wistar rats were randomly divided into A ( n = 24, normal control, with isotonic saline dressing after burns), B ( n = 32, with isotonic saline dressing treatment after hydrofluoric acid burns), C ( n = 32, with wet-dressing treatment after hydrofluoric acid burns), and D ( n = 32, with biological dressing treatment after hydrofluoric acid burns) groups. The rats in the latter 3 groups were inflicted with 3 cm x 3 cm TBSA full-thickness burns, and mortality, concentration of blood calcium , histopathological observation were carried out at 4,8,24 and 72 postburn hours (PBH), with 8 rats at each time-points. In addition, 46 patients with hydrofluoric acid burns were divided into E (with wet-dressing treatment) and F (with biological dressing treatment) groups to compare the curative effect.</p><p><b>RESULTS</b>The mortality in A,B,C,D groups were 0,31.2% ,15. 6% ,6. 2% , respectively. The wound in B group was deepened gradually after burns, but that in D group was slighter when compared with that in C group. The concentration of blood calcium in A group was higher than that in B, C and D groups at each time-points, and that in D groups was higher than that in B and C groups. The concentration of blood calcium in D group at 8 and 24 PBH were [(2.215 +/-0.008) ,(2.216 +/-0.008) mmol/L], which were obviously higher than those in B [(1.813 +/-0.017),(1.912 +/-0.013)mmol/L l] and C [(2.015 +/-0.006), (2.018 +/-0. 010)mmol/L] groups, (P <0. 01). The clinical outcome in E group was much better than that in F group.</p><p><b>CONCLUSION</b>Biological dressing containing calcium and magnesium can be applied in the emergency management and following treatment after hydrofluoric acid burns.</p>

Construction of rat bdnf gene lentiviral vector and its expression in mesenchymal stem cells / 生物工程学报

Recently, mesenchymal stem cells (MSCs) have been one of the target cells of gene engineering. To construct the lentiviral (LV) vectors carrying the brain-derived neurotrophic factor (Bdnf) gene, the rat mesenchymal stem cells (rMSCs) were infected and finally the Bdnf gene-modified rMSCs was obtained. The CDS region of the rat Bdnf gene was obtained with reverse transcriptase-polymerase chain reaction (RT-PCR), and the transfer plasmid (PNL-BDNF-IRES2-EGFP) of the LV vector was constructed. The three plasmids of LV vector: PNL-BDNF-IRES2-EGFP, HELPER, and VSVG were cotransfected to 293T cells to produce the LV vectors, which enabled the coexpression of the Bdnf gene and the enhanced green fluorescent protein (Egfp) gene. rMSCs were separated from the bone marrow of 2-month-old F344 rats, cultured in vitro, and identified. rMSCs were infected by the LV vectors that were produced already and were identified with fluorescent microscope, RT-PCR, immunocytochemical staining, and western blot. The result of sequencing showed that the sequence of the cloned Bdnf gene was consistent with that reported in the GenBank. The PNL-BDNF-IRES2-EGFP plasmid that was identified showed the correct sequence. After the 3 plasmids of LV vectors were cotransfected to the 293T cells, considerable green fluorescence in 293T cells was observed under the fluorescent microscope; the supernatant was collected and concentrated using ultracentrifugation, and the titer of the replication-defective LV vector particles measured was found to be 6.7 x 10(7) TU/mL. After the constructed LV vectors infected the rMSCs, the results obtained using RT-PCR, immunocytochemical staining, and western blot showed that the expression of BDNF in the Bdnf-rMSCs group (experimental group, EG) was significantly higher than that in the PNL-IRES2-EGFP-rMSCs group (mock group, MG) and the rMSCs group (control group, CG) at both mRNA and protein levels. LV vectors carrying the Bdnf gene were constructed successfully. The Bdnf gene-modified rMSCs could express BDNF to a higher degree. This greatly facilitates the next step in the study, such as the long period of therapeutic observation of cerebral ischemia with Bdnf gene-modified rMSCs.

Imaging evaluation of efficacy of radiofrequency ablation treatment for hepatic cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the value of B-ultrasonography, CT and MRI in the evaluation of efficacy of radiofrequency ablation (RFA) for hepatic cancer.</p><p><b>METHODS</b>One hundred patients with hepatic cancer were treated by ultrasound-guided RFA between October 1999 and September 2000. All patients had been examined by serum AFP, B-ultrasound, CT or MRI before and within one month after RFA.</p><p><b>RESULTS</b>Before RFA, 34 patients who had had CT, the tumor showed hypo- or iso-density un-enhancement and enhancement on dynamic scanning. After RFA, 14 patients were examined by CT scan. Compared with the density on CT scan before RFA, 5 patients showed lower density lesion without any enhancement on dynamic scanning, but the other 9 patients showed similar images to the previous CT scan before RFA in some parts of their tumor. Before RFA, 66 patients examined by MRI showed hypo-intensity on T(1)-weighted image, hyper-intensity on T(2)-weighted image and enhancement on dynamic scanning. After RFA, among 86 patients examined by MRI, 44 showed iso- or hyper-intensity on T(1)-weighted image, iso- or hypo-intensity on T(2)-weighted image and no enhancement on dynamic scanning. But 42 patients showed similar images to the previous MRI imaging performed before RFA in some parts of their tumor.</p><p><b>CONCLUSION</b>Both CT and MRI can be used as imaging evaluation tool on the effect of radiofrequency ablation for hepatic cancer. However, MRI is better than CT to detect whether the tumor is necrotic or still partly viable after radiofrequency ablation. Patients can be regarded as clinically cured provided that the serum AFP declines to the normal level from abnormally high level and/or MRI or CT scans show a complete necrotic lesion after RFA.</p>

Percutaneous radiofrequency ablation for liver cancer located in hepatis / 中华外科杂志

<p><b>OBJECTIVE</b>To explore the feasibility, effect and problems of percutaneous radiofrequency ablation (PRFA) performed for small liver cancer located in hepatis.</p><p><b>METHODS</b>Twenty-one patients, who had small primary or metastatic liver cancer confirmed clinically or pathologically that were located in hepatis and less than 5 cm, were performed PRFA between April 2000 and October 2002. All patients were followed up to examine the value of AFP, MRI or CT. Kaplan-Meier estimation was used for the disease-free survival rate and the long-term survival rate.</p><p><b>RESULTS</b>The rate of AFP positive down to negative was 77.8% (7/9). The complete necrosis rate was 90.5% (19/21). The peri-tumor recurrence-free survival rates of 0.5-, 1-, 1.5-, 2-year were all 94.7%. The distant recurrence-free survival rates of 0.5-, 1-, 1.5-, 2-year were 90.0%, 77.1%, 77.1% and 77.1% respectively. The whole survival rates of 0.5-, 1-, 1.5- and 2-year were 89.2%, 82.8%, 82.8% and 55.2% respectively.</p><p><b>CONCLUSIONS</b>Small liver cancer located in hepatis was not the contra-indication of PRFA. If the puncture point and route is selected properly, electrodes outspreaded exactly and the range of heating controlled appropriately, PRFA is an effective method and of less complication rate for small liver cancer located in hepatis. Sometimes, PRFA can be combined with TACE for those tumors of the diameter larger than 3 cm.</p>

Percutaneous hepatic puncture radio frequency ablation in the treatment of hepatocellular carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To observe the curative effect of percutaneous radio frequency ablation (PRFA) on hepatocellular carcinoma.</p><p><b>METHODS</b>Under local or epidural anaesthesia, radio frequency electrodes were percutaneously introduced with ultrasound guide into the tumor in the liver for ablation.</p><p><b>RESULTS</b>Among the 96 hepatic cancer lesions in 60 patients, 41 (42.7%) with diameter < 3.5 cm were ablated by one session. The effective rate (CR + PR) was 100% (41/41) with CR 92.6% (38/41) and PR 7.3% (3/41). During the 6 - 24 month follow up of these lesions, 36 (87.8%) of these 41 patients showed no recurrence by CT or MRI. Fifty-five (57.3%) lesions with diameter 3.5 cm < or = Phi < 12 cm were ablated by two or three sessions, with CR 3.6% (2/55), PR 67.3% (37/55) and CR + PR 70.9% (39/55) by CT or MRI in 1 to 3 months after the treatment.</p><p><b>CONCLUSION</b>Percutaneous radio frequency ablation is effective on liver tumor with Phi < 3.5 cm. It is partly effective on lesions with 3.5 cm < or = Phi < 12 cm.</p>