Comparison of different ureteral length measurement methods in the indwelling of double-J stent after ureteroscopy / 现代泌尿外科杂志

【Objective】 To investigate the application of different ureteral length measurement methods in the indwelling of double-J stent. 【Methods】 Clinical data of 260 patients with double-J stent indwelling after ureteroscopic surgery during Jul.2018 and Dec.2020 were prospectively analyzed. The patients were randomly divided into height calculation group, CT measurement group, KUB group and ureteroscopic measurement group. The length of ureter was calculated accordingly and the appropriate length of double-J stent was selected. KUB was performed on the first day after operation and before extubation to determine the position of double-J stent. The patients completed the ureteral stent-related symptom questionnaire (USSQ), urinary symptom score, lower urinary tract symptom (LUTS) score, pain score, hematuria score, and quality of life score before and after double-J catheter placement. 【Results】 There were no significant differences in age, gender, height, side of stent and urinary symptom score among the four groups (P>0.05). The average lengths of the ureters measured by the four methods were (21.5±1.0) cm, (21.5±1.8) cm, (23.8±1.3) cm and (21.7±1.8) cm, respectively. There were no significant differences among the height calculation group, CT measurement group and ureteroscope group, but there was significant difference between the three groups and the KUB group. The ideal ureteral stent length indwelling ratio in the ureteroscopic group was 76.9%, which was better than that in the other three groups. Postoperative indwelling time was 7-42 d (mean 29.8 d). The USSQ score of the ureteroscopic group before extubation was (14.1±1.5), which was lower than that of the other three groups (P<0.05). The ureteroscopic group was better than the other three groups in the comparison of frequency and urgency of urination, nocturia, hematuria, quality of life score, and pain score (P<0.05). 【Conclusion】 Intraoperative ureteroscopic measurement of the ureteral length is a simple and feasible method in guiding the indwelling of double-J stent to reduce ureteral stent related symptoms.

Three hot topics in the prevention and control of myopia among children and adolescents / 中国学校卫生

Abstract@#In order to improve the prevention and control of myopia, this paper examines the significance of hyperopia reserves and its correct application, identifies problems requiring further attention and sums up past experiences. It puts forward a prevention and control system of myopia after introduction of the physiotherapy instruments, especially those based on the principle of red light. This paper discusses some mechanisms of low concentration atropine eye drops in preventing and controlling myopia. Finally, future research directions are proposed.

Clinicopathological features of second primary lung cancer and pulmonary metastasisin patients with breast cancer / 中华肿瘤杂志

Objective@#To compare the clinicopathological characteristics of second primary lung cancer following breast cancer and lung metastases from breast cancer, and then to analyze the risk factors in breast cancer patients with second primary lung tumor.@*Methods@#Clinical data of 55 breast cancer patients with second primary lung tumor and 205 breast cancer patients with solitary pulmonary metastasis in Shandong Cancer Hospital from January 2006 to January 2017 were retrospectively analyzed. The risk factors of primary lung cancer following breast cancer were analyzed using logistic regression model.@*Results@#Second primary lung cancer in patients with first breast cancer accounted for approximately 21.2%(55/260) of pulmonary malignant solitary nodules, and 0.84%(55/6 580) of all breast cancer patients. The median intervals between the diagnosis of second primary lung cancer or lung metastasis and first breast cancer were 52 months and 42 months, respectively. These two groups showed significant difference between age, time interval between diagnoses, breast tumor size, axillary lymph node metastasis, estrogen receptor, molecular subtype (luminal B and triple-negative) and history of radiotherapy (P<0.05 for all). A multivariate logistic regression model confirmed that age (OR=1.088, P<0.001), breast tumor size(OR=0.480, P<0.001), and radiotherapy history (OR=3.460, P=0.004) were all independent factors for second primary lung cancer.@*Conclusions@#For isolated pulmonary nodules in patients with breast cancer, especially for those with elder age, larger tumor size and radiotherapy history, we should distinguish the second primary lung cancer from pulmonary metastasis. The treatment regimen for lung metastasis and primary lung cancer in patients with breast cancer are entirely distinct. The timely histopathology examinations for pulmonary nodes in patients with breast cancer are recommended.

Application of tumescent anaesthesia in laparoscopic cholecystectomy / 实用医学杂志

Objective To explore the clinical effect of tumescent anaesthesia in laparoscopic cholecystectomy(LC). Methods We analyzed the data of 71 patients who underwent LC for gallbladder benign disease. Anesthesia swelling fluid was injected in 37 cases before the operation in the gallbladder triangle( group D)and another 34 cases received no injection(group C). SBP,DBP,HR and Spo2 were monitored at 4 min before gallbladder traction(T1),1 min(T2),6 min(T3)and 11 min after gallbladder traction(T4),and when the gallbladder was dissected from gallbladder bed completely(T5). The operation time,intraoperative blood loss,postoperative exhaust time,time of getting out of bed after the surgery and length of stay were recorded. The gallbladder rupture, vascular injury ,bile duct injury ,intestinal injury ,placement of drainage tube ,intra-abdominal hemorrhage , intraoperative use of atropine or ephedrine and nausea and vomiting 24 hours after the operation were recorded. Results In group C,SBP,DBP and HR were significantly decreased at T2-T4 when compared with those at T1, and were significantly lower when compared with those in group D(P < 0.05). There was no significant difference in oxygen saturation between the two groups(P > 0.05). There were shorter operation time and less intraoperative blood loss in group D when compared with those in group C and the difference was statistically significant (P <0.05). The anus exhaust time,time of getting out of bed after the surgery,length of stay,intraoperative intra-abdominal hemorrhage and nausea and vomiting in the two groups indicated no statistical significance (P > 0.05). Gallbladder damage,vascular injury,placement of drainage tube and the incidence of adverse event were less in group D than those in group C. Conclusion Swelling anesthesia in LC can shorten the operation time ,induce less postoperative complications and maintain blood circulation stability.

Comparative study on the efficacy and safety between pegfilgrastim (PEG-rhG-CSF) and recombinant human granulocyte colony-stimulating factor in promoting hematopoietic recovery after allogeneic hematopoietic stem cell transplantation after hematological malignancy / 中华血液学杂志

Objective@#To observe the efficacy and safety between Pegfilgrastim (PEG-rhG-CSF) and Recombinant human granulocyte colony stimulating factor (rhG-CSF) in hematological malignancy after allogeneic hematopoietic stem cell transplantation (allo-HSCT) .@*Methods@#157 patients after allo-HSCT were enrolled in this study from June 2015 to November 2016. Two agents of G-CSF were used to stimulate hematopoietic recovery after transplantation. There were 65 cases in PEG-rhG-CSF and 92 cases in rhG-CSF groups. Patients in PEG-rhG-CSF group were given a single subcutaneous dose of 6 mg on the first day and +8 d, while cases in rhG-CSF group were given in dose of 5 μg·kg-1·d-1 by subcutaneous injection from +1 d continuing to neutrophils more than 1.5×109/L, and then the indicators and survival rates in two groups after transplantation were compared.@*Results@#①There were no significant differences of the neutrophil implantation time[13.5 (8-12) d vs 13 (9-24) d, P=0.393] and platelet implantation time [14 (9-160) d vs 14 (9-92) d, P=0.094] between PEG-rhG-CSF and rhG-CSF groups respectively. There were no significant differences in terms of neutropenia period (P=0.435) , number of cases who got fever during neutropenia (P=0.622) , and the median time of fever in neutropenia period (P=0.460) , respectively between the two groups. There were no significant differences of erythrocyte and platelet transfusions (P=0.074, P=0.059) within 1 month after transplantation. ②There were no significant differences with regard to the incidences of acute GVHD[23.1% (15/65) vs 34.8% (32/92) , P=0.115], chronic GVHD[20.0% (13/65) vs 32.6% (32/92) , P=0.081], Ⅱ-Ⅳdegree of acute GVHD[30.0% (13/65) vs 30.4% (30/92) , P=0.287] and extensive chronic GVHD[9.2% (6/65) vs 20.7% (19/92) , P=0.135] between PEG-rhG-CSF and rhG-CSF groups. ③There were no significant differences in terms of disease free survival (DFS) (62.5% vs 61.4%, P=0.478) and overall survival (OS) (67.4% vs 67.3%, P=0.718) between PEG-rhG-CSF and rhG-CSF groups. ④There was no significant difference of the non-relapse mortality (NRM) between PEG-rhG-CSF and rhG-CSF groups[20.5% (95%CI 11.4%-37.0%) vs 32.6% (95%CI 22.2%-47.9%) , P=0.141]. The relapse rate was not statistically significant[14.9% (95%CI 7.4%-29.8%) vs 10.0% (95%CI 5.0%-20.0%) , P=0.299].@*Conclusion@#Compared with rhG-CSF, PEG-rhG-CSF could reduce the times of injection. There were no differences in terms of hematopoietic recovery, the incidence of GVHD, relapse rate, DFS and OS rates after allo-HSCT between two groups.

Clinical efficacy of recombinant activated factor Ⅶ a for 16 hematonosis with moderate or severe bleeding / 中华血液学杂志

Objective@#To analyze the efficacy of recombinant activated factor Ⅶ a (rF Ⅶ a) on hematonosis with moderate or severe bleeding signs.@*Methods@#Of total 16 cases with rF Ⅶ a treatment from May 2013 to May 2016, 8 cases received allogeneic hematopoietic stem cells transplantation (allo-HSCT) and the other were non-transplantation patients. In two groups, there was no significant difference on rF Ⅶ a usage and dosage. 15 patients with acute graft-versus-host disease (aGVHD) after allo-HSCT were control group (without rF Ⅶ a) .@*Results@#①The total response rate was 75.0% (6/8) in non-transplantation group and 37.5% (3/8) in transplantation group, respectively. Median interval for hemorrhage stop was 38.5 hours in non-transplantation group and 63.0 hours in transplantation group. The median overall survival (OS) was 201.0 and 29.0 days for non-transplantation group and transplantation group, respectively, and the OS rate was 50.0% (4/8) and 25.0% (2/8) , respectively. The bleeding-related mortality rate was 50.0% (2/4) and 83.3% (5/6) , respectively. ②Of the 16 cases, 9 showed response to rF Ⅶ a treatment and the other 7 cases’bleeding signs did not alleviate. The median OS was 268.0 in 9 cases with response and 24.0 days in 7 cases without response, respectively. ③In patients with intestinal aGVHD complicated with intestinal hemorrhage, the median OS of observation group (n=6) and control group (n=15) were 25.5 days and 20.0 days, respectively.@*Conclusion@#Patients with hematological diseases, especially patients after allo-HSCT, had high bleeding-related mortality, and rFⅦa therapy had a obvious hemostatic efficacy. The survival rate of patients with response was higher than that of cases without response. The causes of poor hemostasis efficacy of rF Ⅶ a therapy were associated with unsatisfactory control of complications in patients with intestinal bleeding after allo-HSCT.

Impact of essential medicine system on rational drug use in Community Health Care Center of Chun′an District / 药学实践杂志

Objective To study the impact of essential medicine system on rational drug use in Community Health Care Center of Chun′an District .Methods Essential drug system in Community Health Care Center of Chun′an District in Zhejiang Province was taken as the intervention group ,and before implementation was taken as contrast group ,the essential medicin system was analyzed whether it was or not to promote the rational use of drugs by prescription surveys .Results Average num-ber of drugs per encounter was lower from 2 .68 to 2 .23(P< 0 .05) ,average expenditure per encounter was lower from 42 .11 yuan to 37 .75 yuan(P<0 .05) ,the use of essential medicines was riser from 79 .90% to 89 .81% (P<0 .05) ,the three in-dexes had significant difference ,the percentage of injections per encounter was lower from 37 .87% to 28 .50% (P>0 .05) ,the percentage of antibiotics per encounter was lower from 36 .52% to 24 .46% (P> 0 .05) ,the two indexes had no significant difference .Conclusion After execution of essential medicine system in Community Health Care Center of Chun′an District ,av-erage number of drugs and average expenditure per encounter were lower and the use of essential medicines was riser ,but the percentage of antibiotics and percentage of injections were not been improved ,belonged to irrational category ,although there had essential medicines manual ,but effective promoting measures still was deficient .

A preliminary determination of foot-related tissue elastic modulus / 中国组织工程研究

al from abroad, which have no reports in China. METHODS: The dissection of flexor policis longus tendon and flexor policis brevis muscle and the medial and extensor halucis longus, flexor policis longus, adductor muscle and abductor halucis muscle cross head and oblique head, medial and lateral head of flexor policis brevis muscle and flexor halucis longus tendon and the extensor halucis longus tendon. These parameters included length, width, thickness, cross-sectional area, lateral heads, extensor halucis longus muscle and tendon and the transverse head of adductor policis muscle and the oblique head, abductor policis brevis from the left leg and foot of fresh female specimens was performed. The cross-sectional area and length located in a fixture were measured and calculated for each sample. Sample loading was done, and one sample was measured four times to gather strength limit, maximum load data, and the load displacement curve. According to Hooke’s law, the elastic modulus of each specimen was calculated. al from abroad, which have no reports in China. METHODS: The dissection of flexor policis longus tendon and flexor policis brevis muscle and the medial and extensor halucis longus, flexor policis longus, adductor muscle and abductor halucis muscle cross head and oblique head, medial and lateral head of flexor policis brevis muscle and flexor halucis longus tendon and the extensor halucis longus tendon. These parameters included length, width, thickness, cross-sectional area, lateral heads, extensor halucis longus muscle and tendon and the transverse head of adductor policis muscle and the oblique head, abductor policis brevis from the left leg and foot of fresh female specimens was performed. The cross-sectional area and length located in a fixture were measured and calculated for each sample. Sample loading was done, and one sample was measured four times to gather strength limit, maximum load data, and the load displacement curve. According to Hooke’s law, the elastic modulus of each specimen was calculated. Abstract BACKGROUND:Currently, the material parameters of foot three-dimensional finite element models are almost OBJECTIVE:To preliminarily measure the parameters of foot muscle and tendon materials in Chinese people. RESULTS AND CONCLUSION: Relevant measurement data were harvested from nine samples, including the maximum loading, ultimate strength and elastic modulus test.

Effect of recombinant human p53 adenovirus (Ad-p53) combined with EGFR inhibitor gefitinib on the sensitivity of breast cancer MDA-MB-468 cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To observe the impact of concurrent administration of recombinant human p53 adenovirus (Ad-p53) with EGFR inhibitor gefitinib on breast cancer MDA-MB-468 cells.</p><p><b>METHODS</b>MDA-MB-468 cells were treated with Ad-p53 and/or gefitinib. The effect of Ad-p53 and gefitinib on the growth of MDA-MB-468 cells was evaluated by MTT assay. Cell apoptosis was detected by flow cytometry. Western blot analysis was used to detect the alteration of p53,EGFR, phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway and apoptosis-related proteins. Ad-p53 combined with gefitinib was used in vivo to explore their effect on tumor xenograft in nude mice. Immunohistochemistry was used to detect the p53 expression in vivo.</p><p><b>RESULTS</b>The MTT assay showed a stronger inhibitory effect of gefitinib on MDA-MB-468 cells infected with Ad-p53 than on the control cells. Cell apoptosis assay revealed that the apoptosis rates of MDA-MB-468 cells in vehicle-treated group, Ad-p53 group, gefitinib group, and combination group were 8.5%, 17.4%, 20.5% and 32.6%, respectively. The apoptosis rate of MDA-MB-468 cells in the combination group was higher than that in other groups (P < 0.05, for all) . Western blot analysis revealed that the expression of p53 was significantly up-regulated in the presence of Ad-p53. The combination of Ad-p53 and gefitinib significantly down-regulated p-Akt (S473)(P < 0.01) and up-regulated caspase-9 and cleaved caspase-3 (P < 0.01 for both). Tumor inhibition rates (TIR) in the Ad-p53, gefitinib, and combination groups were 35.7%, 28.7% and 74.4%, respectively. Ad-p53 and gefitinib combination therapy significantly reduced the tumor burden in nude mice (P < 0.05 for all).Immunohistochemistry showed that after intratumoral administration of Ad-p53, wild-type p53 was increased (P < 0.01). p53 expressions in the vehicle-treated, Ad-p53, gefitinib and combination groups were 45.2%, 80.1%, 50.7% and 90.6%, respectively.</p><p><b>CONCLUSIONS</b>Wild-type p53 may reverse the sensitivity of MDA-MB-468 cells to gefitinib through down-regulation of the PI3K/Akt pathway. The apoptotic activity induced by this combination treatment might be regulated through caspase cascade.</p>

An applied anatomical study of the right posterior sectional Glissonean pedicle / 中华肝胆外科杂志

Objective To study the anatomical structure and the distribution of the right posterior sectional Glissonean pedicle so as to provide the anatomical basis for liver resection of the right posterior section.Method By studying the morphological features of the right posterior sectional Glissonean pedicle through peeling and dissecting 20 adult corpses without liver pathological.Result In 95％ of the right posterior sectional Glissonean pedicles there were shared branches.The angle between the GRL/GRA was 81.4° ± 13.8°.In 30％,the right posterior sectional Glissonean pedicle presented as a Y or a V.In a Y shaped,the angle between G6/G7 was 71 ° ±3.92°.The shapes of 70％ of the right posterior sectional Glissonean pedicle presented as a C.The number of branches at the tertiary level was between 3-8.Conclusion The distribution of the right posterior sectional of the Glissonean pedicle did not entirely fit the description of Couinaud's segments.Obvious individual differences existed.Under this circumstance,understanding the anatomy of the right posterior sectional Glissonean pedicle before carrying out partial hepatectomy would be helpful in reducing surgical risks and postoperative complications.

Research progress of hemodynamic signal and liver regeneration / 中华消化外科杂志

Liver has great ability in regeneration,and liver regeneration is closely related to the prognosis of patients who received liver surgeries.The mechanisms of liver regeneration include biochemical theory and hemodynamic theory.Portal perfusion with moderate pressure is an important factor for starting liver regeneration.Hepatic artery buffer response,fluid shear stress and gasotransmitter play important roles in the liver regeneration.Learning the mechanisms of hemodynamic signal is of great importance for the treatment of liver diseases.

Anatomical symmetry of the intrahepatic Glisson system / 解剖学报

Objective To observe the anatomical symmetry of the structures and istribution of the Glissonean pedicle of the intrahepatic Glisson system ,integrating with embryology and comparative anatomy .Methods Morphology of the Glissonean pedicle of liver was examined through peeling and dissecting 20 adult corpses without liver pathological changes.The relevant data were collected and analyzed statistically .Meanwhile, we tried to elucidate elaborating the symmetry theory of liver anatomy through the dissection anatomy ,embryonic anatomy and comparative anatomy .Results The angle between main stem of Glisson system/left branch of Glisson system(GM/GL) was (76.7 ±17.36)°.The angle between GM/GR was (81.4 ±13.8)°.The length of the the Glisson pedicle of left hepatic was (3.1 ±0.76) cm.The length of the the Glisson pedicle of middle hepatic was ( 2.61 ±0.72 ) cm.The length of the the Glisson pedicle of right hepatic was (1.5 ±0.50)cm.The shapes of the Glissonean pedicle stem of the left hepatic presenting arch , the number of radial level 3 branches were between 2-8.The shapes of the Glissonean pedicle stem of the middle hepatic continuing the main of Glissonean pedicle , the number of radial level 3 branches were between 2-6.The shapes of 30%of the Glissonean pedicle of the right hepatic presenting Y and V , 70% of the Glissonean pedicle of the right hepatic presenting C , the number of radial level 3 branches were between 3-8.Conclusion In the light of morphology ,embryology and comparative anatomy, it is reasonable to divide the liver into left ,middle,right lobe by Glissonean pedicle of radial level 2 branches and the liver is an axiality and symmetry organ .

Effects of finasteride therapy on the mRNA expressions of HSP 70, HSP 22 and levels of TNF-α, IL-1β in patients with benign prostatic hyperplasia / 中华老年医学杂志

Objective To explore the effects of finasteride therapy on the expressions of heat shock protein 70(HSP70)mRNA,heat shock protein 22 (HSP22)mRNA,tumor necrosis factor-alpha (TNF-a),interleukins-1β (IL-1β) in elderly patients with benign prostatic hyperplasia (BPH).Methods Totally 68 elderly men patients with BPH were divided into two groups:regular treatment group (n=34) and finasteride treatment group(n=34),30 elderly men patients without BPH were as control group.The blood samples of patients after treatment were collected for the detections of HSP70mRNA and HSP22mRNA with RT-PCR,meanwhile,the levels of TNF-α,IL-1β were measured by radioimmunoassay (RIA).Effect of treatment was evaluated in accordance with International Prostate Symptom Score (IPSS).Results The expressions of HSP70 mRNA and HSP22 mRNA,TNF-α,IL-1β in BPH patients were higher in finasteride group than in control group (P＜0.01).After 12 weeks therapy,the levels of HSP70mRNA,HSP22mRNA,TNF-α and IL-1β were decreased in finasteride group (P＜0.01 or P＜0.05) but no significantly difference in regular treatment group.Compared with regular treatment group,the expressions of HSP70 mRNA,HSP22mRNA and TNF-α were reduced in the finasteride group after treatment (P＜0.01 or P＜0.05),IPSS was more improved (P＜0.05),but IL-1β was not distingished between two groups (P＞0.05).IPSS was positively related to the levels of HSP70 mRNA,HSP22 mRNA,TNF-α and IL1β (r=0.4251,0.4976,0.4562,0.3627,P＜0.01 or P＜0.05).Conclusions The occurrence and development of BPH are closely related with immunoinflammatory factors.Finasteride not only affects the levels of dihydrotestosterone but also can inhibit the expression of immunoinflammatory factors to improve symptom by reducing the volume of benign prostatic hyperplasia.

New understanding and research progression for triple-negative breast cancer / 国际肿瘤学杂志

Triple-negative breast cancer (TNBC) is a heterogeneous disease.It has distinct risk factors,molecular biology features,clinical presentations and prognosis.TNBC recurrence is common after resection,and the survival rate is low and available treatment options are few after recurrence.To date,chemotherapy is the main treatment strategy for TNBC.There is a great need for new molecular predictive marks and drug targets for improving the efficacy of TNBC treatment.

Breast cancer molecular subtypes and its clinical significance / 国际肿瘤学杂志

Breast cancer is regarded as a heterogenous disease classified into different molecular subtypes,resulting in different biologic behavior and sensitivity to treatment respectively.Molecular biology typing that based gene expression profiles and biological characteristics of breast cancer can reflect the biological behavior of tumors exactly,which has important clinical significance.

The effect of NGF gene modified Scwhann cells on DRG neurons repair after compressed injury / 中华骨科杂志

ObjectiveTo study the effect and mechanism of NGF gene modified SCs on DRG (dorsal root ganglion) neurons repair after compressed injury.MethodsSCs were obtained by one enzyme digestion method.SCs were transfected with NGF gene by adenovirus.Thirty-two female SD rats with compression injury of dorsal root ganglia on right lumbar nerve roots.were divided randomly into following groups:Normal saline(NS) group,Pure SCs group,Ad-NGF group,and SCs+NGF group.Nerve root tissues were harvested 2 weeks after treatment.Western blot were used to detect the proNGF volume in nerve root tissue lysis; Double-labeling fluorescent Immunohistochemistry(IHC) was used to count the number of β-Tubulin Ⅲ positive cells and activating transcription factor 3 positive cells.The ratio of injured neurons to survived neurons was calculated.ResultsWestern bolt showed the proNGF volume in nerve root tissue lysis of SCs+NGF group increased dramatically.Double-labeling fluorescent IHC showed SCs+NGF group vs any group,the density of survived DRG neurons(β-Tubulin Ⅲ positive cells) increased significantly,meanwhile the percentage of injured neurons (ATF3 positive cells) in survived neurons decreased dramatically.Conclusion NGF gene Modified SCs could promote the survival of DRG neurons after compression injury and decrease the ratio of injured neurons.We conclude that this study provides a new treatment strategy for the patients who suffer from chronic compression injury on nerve roots and DRG neurons.

Effect of different cell seeding concentrations on chondrogenic differentiation of adipose derived sromal cells in three-dimensional culture / 中国组织工程研究

BACKGROUND: The implanted cartilage calls can synthesize cartilage matrix as cartilage in cartilage tissue enginsedng, and the density of implanted cells is the key point.OBJECTIVE: To evaluate the effect of cell seeding concentration on the chondrogenic differentiation of the adipose dadved sromal cells (ADSCs).DESIGN, TIME AND SETTING: The in vitro cellular-scaffold observation was performed at the cytobiological laboratory of China Medical University from November 2007 to July 2008.MATERIALS: Six male SD rata with clean grade were supplied by the Experimental Animal Center of China Medical University.METHODS: Totally 5 g/L type ; collagen solution and 20 g/L chitosan was mixed in a mould with volume ratio of 7:3, after lyophillization, it was cut into pieces with 5 mm ～ 5 mm x 2 mm, followed by crosslinking with ethanol contained of 2% chondroitic acid at room temperature. After washing with double distilled water and freeze drying, the chitosan-collagen-chondroitin sulfate copolymar matrices scaffolds were harvested. ADSCs isolated from rat inguinal fat pads were digested with collagenase and trypsase. The prepared scaffolds were randomly divided into 3 groups, and the third passage cells with density of 2×10 9/L,2×10 109/L, and 2×10 11/L were seeded into chitosan-coflagen-chondroitin sulfate scaffolds, and cultured in chondrogenic medium for 3 weeks.MAIN OUTCOME MEASURES: The expression of cartilage specificity gene was detected by hematoxylin-eosin staining, type Ⅱ collagen immunohistochemical staining and RT-PCR.RESULTS: Hematoxylin-eosin staining showed that after 3 weeks of culture, the cell proliferated and differentiated well, especially in 2x101～/L group, more extrocelluer matrices were produced and cartilage lacuna-structure could be seen. The type Ⅱ collagen was positive expressed in each group, which showed a gradually increasing tendency with the cell seeding concentration increasing. RT-PCR showed that the expression of proteoglycen and type Ⅱ collagen mRNA were slowly increased. However the expression of Ⅹ collagen mRNA was decreased with increasing cell seeding concentration.CONCLUSION: The chitosan-collagen-chondroitin sulfate copolymer matrices can provide an appropdate environment for the generation of cartilage-like tissues and high call seeding concentration of 2×1010/L facilitate ADSCs to differentiate into cartilage.

TIMP-1 secreted by fibroblasts inhibits tumor cell invasion and metastasis in mouse melanoma / 生物医学工程学杂志

We constructed a recombinant adenoviral vector expressing human tissue inhibitors of metalloproteinase-1(TIMP-1), and evaluated the inhibition of TIMP-1 secreted by primary fibroblasts after infection with adenovirus-mediated TIMP-1 gene (Ad-TIMP-1) on tumor cell invasion and metastasis in mouse melanoma. It was found that TIMP-1 was detected in the supernatants of cultured mouse primary fibroblasts after infection with Ad-TIMP-1 by indirect enzyme-linked immunosorbent assay (ELISA). The TIMP-1 secreted by Ad-TIMP-1 infected primary fibroblast significantly inhibited B16BL6 cell invasion and metastasis both in vitro and in vivo. We also demonstrated that the primary fibroblasts transfected by Ad-TIMP-1, after being subcutaneously injected into mouse, can secreted TIMP-1 into the blood of mouse and maintained at the therapeutic in vivo levels of TIMP-1. These results suggest that the preparation of Ad-TIMP-1 infected primary fibroblast be an effective method to deliver TIMP-1 gene in vivo, which provides a new strategy of gene therapy and has the potential for clinical applications in the treatment of tumor cell metastasis.

Cartilage tissue engineering by collagen-chitosan-chondroitin sulfate scaffold seeded with rat adipose tissue-derived stromal cells in vitro / 中华骨科杂志

Objective To evaluate the character of the collagen-chitosan-chondroitin sulfate scaffold seeded with rat adipose tissue-derived stromal cells. Methods A dipose tissue were harvested from 6 weeks old Wistar rats and the stromal cells were harvested by type Ⅰ collagenase and then cultured in vitro. Type Ⅰ collagen was fully mixed with chitosan, freeze-dried and cross-linked with chondroitin sulfate, then freeze-dried again and sterilized by ethylene oxide. The pore diameter, water content, porosity of the scaffold were tested. The adipose tissue-derived stromal cells were digested, seeded into the plates, scaffold, and cen-trifuged into pellet, and then induced into cartilage. MTT detection for cell proliferation was done. After 3 weeks, the cell morphology, and cell proliferation and adhesion were observed, and chondrngenic differenti-ation was also analyzed. Results The pore diameter, water content, porosity tested for the scaffold showed an appropriate form. Cell proliferation showed faster in the scaffold and pellet culture system after 5 day, there was still cell proliferation in the scaffold system after 14 days but no obvious changes in the pellet cul-ture system; ceils on the scaffold proliferated densely showed by histological staining, but there was a scaf-fold structure residues in the inner layer. The finding of type Ⅱ immunohistochemistry stain showed that cells express strong positive for type Ⅱ collagen in the scaffold and pellet culture system whereas it was weakly positive in the plate culture system; the specific mRNA for cartilage, type Ⅱ collagen, aggrecan and SOX-9 were expressed in all three systems showed by RT-PCR, but type X collagen was expressed continu-ously in the plate culture system and expressed after 21 days in the pellet culture system, whereas it was not detected in the collagen-chitosan-chondroitin sulfate scaffold system. Conclusion The parameters of the collagen-chitosan-chondroitin sulfate scaffold were suitable in our study. The results suggested that it can promote the adipose tissue-derived stromal cells proliferation and chondrogenic differentiation better than the plate and pellet culture systems and maintain the phenotype of chondrocytes well; it is the optimal choice for cartilage tissue engineering in the future.

Association of three cytokines with graft versus host disease after allogeneic hematopoietic stem cell transplantation for leukemia / 中国组织工程研究

BACKGROUND: Recently, a few studies have reported the correlation between transforming growth factor-α (TGF-α) and graft-versus-host disease (GVHD); however, the combination of TGF-α with other cytokines in patients with chronic or acute GVHD requires further study.OBJECTIVE: To analyze the changes of serum tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4), and transforming growth factor-α (TGF-α) in leukemic patients after allogeneic hematopoietic stem cell transplantation (Allo-HSCT) and investigate the effects of these cytokines on different grades of GVHD.DESIGN: Case control study.SETTING: Department of Hematology, Organ Transplantation Center, the Second Affiliated Hospital, General Hospital of Chinese PLA; Department of Nuclear Medicine, the Second Affiliated Hospital, General Hospital of Chinese PLA.PARTICIPANTS: Forty-two leukemic patients (23 males and 19 females, 16-68 years old, mean age of 35 years) who underwent Allo-HSCT for the first time were selected from the Department of Hematology, Organ Transplantation Center, the Second Affiliated Hospital, General Hospital of Chinese PLA and Department of Transplantation, the 307 Hospital of Chinese PLA from June 2005 to June 2007. Twelve patients had acute granulocytic leukemia (AGL), fifteen patients had acute lymphocytic leukemia (ALL), and fifteen patients had chronic granulocytic leukemia (CGL). Among the 42 patients, 37 underwent peripheral blood transplantation and five received bone marrow transplantation. Twenty-one patients had acute GVHD (18 cases in grades Ⅰ-Ⅱ and three cases in grades Ⅲ-Ⅳ) after Allo-HSCT, but the other 21 patients did not. Fourteen patients had chronic GVHD (five cases of limited type and nine cases of extensive type), but the other 28 patients did not. An additional 30 healthy subjects (18 males and 12 females, 20-70 years old, mean age of 44 years) were collected as a normal control group. All patients provided confirmed consent, and the study was approved by the local ethics committee.METHODS: Levels of serum TNF-α, IL-4, and TGF-α in leukemic patients with Allo-HSCT and normal subjects were measured by radio-immuno-assay, the cytokines levels of the patients with/without acute GVHD, of those with/without chronic GVHD and of different grades of GVHD were compared.MAIN OUTCOME MEASURES: Comparisons of serum TNF-α, IL-4, and TGF-α among the groups.RESULTS: All 42 leukemic patients and 30 healthy subjects were included in the final analysis. Levels of TNF-α, IL-4, and TGF-α in patients with acute or chronic GVHD were significantly higher than those in the normal subjects (P<0.05-0.01). Levels of TNF-α and IL-4 in patients without acute GVHD were significantly higher than those in the normal subjects (P<0.01,0.05). Levels of TNF-α, IL-4, and TGF-α in patients with acute GVHD were significantly higher than those in patients without acute GVHD (P<0.05). Levels of TNF-α, IL-4, and TGF-α in patients with chronic GVHD were significantly higher than those in patients without chronic GVHD (P<0.05). Levels of serum TNF-α and TGF-α in patients with acute GVHD of grades Ⅲ-Ⅳ or chronic GVHD of extensive type were significantly higher than those in patients with acute GVHD of grades Ⅰ-Ⅱ or chronic GVHD of limited type (P<0.05-0.01).CONCLUSION: After Allo-HSCT, dynamically monitoring changes of levels of TNF-α, IL-4, and TGF-α may serve as a possible means of predicting the onset of acute or chronic GVHD and may contribute considerably to deciding clinical severity of GVHD.