Objective To explore the mechanism of protection role of ambroxol against acute lung injury (ALI)by studying the change of cytokine mRNA expression during the course of ALI.Method The study was composed of experiments both in vivo and in vitro. (1)Experiments in vivo were as follows.Twenty-four sprague-Dawley rats were randomly divide into 3 groups,namely,control group(n=8),acute lung injury group(LPS group,n=8)and ambroxol group(LPS+A group,n=8).The rat model of acute lung injury was induced by intraperitoneal injection of 10 mmol/L lipoopolysaccharide(LPS).The pathological alteration of lung tissue and arterial partial pressure of oxygen(PaO2)were observed.Expressions of TNF-α,IL-10 and IL-24 mRNA were determined by using RT-PCR assay. (2)Experiments in vitro were the followings.Alveolar macrophage cells were collected and divided into 3 groups as above mentioned.Cells were treated with normal saline,with LPS,and with LPS plus ambroxolin dose of 180 μmol/L at 0,6,12 and 24 hours after exposure of LPS,respectively,in 3 groups as above stated.The expressions of TNF-α,IL-10 and IL-24 mRNA were also determine by using RT-PCR.Results The pathological changes of could be parially ameliorated by giving ambroxol.Massive hemorrhage along with vascular edema and infiltration occurred in the lungs of ALT rats.The pathological alterations in ALI rats treated with ambroxol were less severe.The expressions of TNF-α,IL-10 and IL-24 mRNA were dramatically increased in ALI rats,and were partially attenuated after treatment of ambroxol.Macrophages oxposed to LPS for 6 hours showed dramatical increase in expressions of TNF-α,IL-10 and IL-24mRNA those remained at high levels afterwards.The expressions of TNF-α,IL-10 and IL-24 mRNA in macrophages after exposure to both LPS and ambroxol were increased less than those exposed to LPS alone.Conclusions Ambroxol can partially ameliorate the expressions of TNF-α,IL-10 and IL-24 mRNA in alveolar macrophage induced by LPS.

Objective:To investigate the values of immunophenotype and the Collagen type1 alpha1/Proto-oncogene Proteins c-sis (COL1A1/PDGFB) fusion gene in the diagnosis of dermatofibrosarcoma protuberans (DFSP). Methods:IHC markers and the COL1A1/PDGFB fusion gene were detected by IHC staining and interphase fluorescence in situ hybridization (FISH) in 73 cases previously diagnosed as DFSP. A total of 85 and 10 non-DFSP cases were also included as controls for IHC staining and FISH, respectively. Results:In the 73 DFSP cases, the positive detection rates for immunohistochemical marker vimentin, CD34, CD99, S100, desmin and SMA were 100%, 91.78%, 61.64%, 0, 0, and 6.85%, correspondingly. Protein expression levels in these cases varied from the control group, and CD34 ex-pression was significantly different among the differential diagnoses. The positive detection rate for the COL1A1/PDGFB fusion gene was 86.96%(60/69), whereas the gene expression in the control group was negative. Conclusion:The COL1A1/PDGFB fusion gene is a highly specific and sensitive marker in the diagnosis of DFSP. CD34 is a suitable marker for DFSP.

Objective To study the protection of taurine(TAU) against apoptosis of neurons induced by manganese(Mn) in vitro. Method Cortex neurons were separated from Wistar neonatal rats and cultured in vitro.The assays began when neurons grew under the best conditions. Cells were randomly divided into 7 groups: control group,Mn-added groups (Mn 0.2,0.6 and 1.0 mmol/L respectively),TAU-intervened groups (1.5mmol/L TAU with Mn). All treatments lasted 24h. Terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) was used to observe the morphology of apoptosic neurons. Flow cytometry (FCM) was used to quantitate neuron apoptosic rates. Results (1) Typical morphologic charateristic was found in Mn-added groups. TAU intervention could protect against the effect of 1.5mmol/l Mn on neurons. (2) FCM indicated that TAU can protect against neurons apoptosis induced by Mn. Conclusion Taurine can protect neurons from apoptosis induced by Mn in vitro.

BACKGROUND: Neural stem cells are a kind of special cells possessing self-renewal and multi-directional differentiation potential. They are ideal carriers for studying the occurence, development and development rule of nervous system and their inherent regulation mechanism of molecular biol ogy and cytology. They have wide applicative prospect in the treatment of nervous system disease and injury. Investigating a set of convenient, effective and useful culture system of neural stem cells cultured in vitro is the prerequisite and basis for application. OBJECTIVE: To observe the effect of serum culture medium containing basic fibroblast growth factor on the in vitro culture of neural stem cells of mice. DESIGN: Single-sample observation. SETTING: Department of Histology and Embryology, Youjiang Nationality Medical College. MATERIALS: Five 2-month-old Kunming white mice of either sex were used in this experiment. METHODS: This experiment was carried out at the Department of Histology and Embryology, Guangxi Medical University from June 2003 to May 2005. ① The cerebral cortical cells were isolated from cerebral cortex and cultured in vitro of the adult mouse cerebral cortex and made into cell suspension , then put in the DMEM/F12 (1:1)culture medium (containing fetal bovine serum of 0.15 volume fraction, basic fibroblast growth factor of 20 μg/L, penicillin of 100 u/mL and streptomycin of 100 u/mL ) .The attachment culture method was used to acquire the clone cells. ② Expression of nestin antigen of clone cells was detected with immunohistochemical technique. MAIN OUTCOME MEASURES: ① Morphological observation of cultured cells. ② Observation of cultured cells stained by haematoxylin and eosin . ③ Identification of neural stem cells. RESULTS: ① Morphological observation of cultured cells: After inoculation, most of the cells attached to the wall within 12 hours, cells were thin and flat. 3 days after inoculation, cells began to grow and proliferate. There were several scores to several hundreds of cell clones on the 5th day after inoculation and cells covered 80%-90% of the bottom of the bottle on the 7th to 8th days. After passage, the cells still grew adhesively with regular cellular morphology and clear borderline. ② Observation of cultured cells stained by haematoxylin and eosin: It was shown that the cells presented round or ellipse. There was little cytoplasm, presenting acidophily. Nucleus was big and round, single in the center, deep stained.③ Identification of neural stem cells: Immunofluorescent detection showed that cell nestin antigen positive. CONCLUSION: After cultured in serum culture medium containing basic fibroblast growth factor, cerebral cortex cells possess very strong reproductive activity and express nestin antigen, still possessing the characteristics of neural stem cells.

Objective High expression of multi-resistant transporter ATP-binding cassette super family G member 2 (ABCG2) is a major cause of drug resistance and chemotherapeutic failure of cancer .This study was to investigate the significance of ABCG2 expression in adrenocortical cancer cells after cyclophosphamide ( CTX) intervention in vivo . Methods Ten male and fe-male BALB/C-nu mice were randomly divided into a cyclophosphamide ( CTX) group and a control of equal number .SW-13 cells were subcutaneously injected into the nude mice to establish a model of subcutaneous transplantation tumor , followed by intraperitoneal injec-tion of CTX and isotonic saline solution into the two groups of mice , respectively .Then the expression of ABCG 2 in tumor tissue and primarily cultured cells was detected by immunohistochemistry and flow cytometry . Results The expression of ABCG 2 in the tumor tissue was significantly higher in the CTX than in the control group ([69.1 ±1.83]%vs [53.4 ±1.65]%, P<0.05), and so was that in the primarily cultured cells ([97.89 ±1.36]% vs [81.88 ±8.31]%, P<0.05). Conclusion The ABCG2 gene is in-volved in the drug resistance of adrenocortical carcinoma and may be a therapeutic target of the malignancy .

Objective To investigate the occurrence of apoptosis and its relationship with endocrine hormones altemtions in rats with acute obstructive pancreatitis(AOP). Methods The model of AOP wag establisbed by ligation of pancreaticobiliary duct.8,12 hrs after operation,the serum insulin,glucagons and amvlase were determined;pancreatic tissues were harvested and apoptotic rate wag evaluated by TUNEL and flow cytometry(Annexin V-FITC/PI assay).Results 8 and 12 hrs after AOP induction,serum amylase levels wefe(1198±687)U/L and(1698±1103)U/L respectively;serum insulin levels were(8.1±5.8)ng/ml and (12.7 ±6.9)ng/ml respectively;sertlm glucagon levels were(6.8±4.6)ng/ml and(7.3±2.9)ng/ml respectively;all these parameters were significantly high than(404±222)U/L,(5.6±2.7)ng/ml and(2.6±2.1)ng/ml in the sham operation group(P<0.05).AnnemnV FITC/PI assay confirmed apoptosis occurred both in exocrine acinus cells and endocrine panclreas islet;and the apoptotic rate wag(20.5±11.2)%and (15.5±8.9)%at 8 and 12 hrs after AOP induction,which wag significantly high than(4.2±1.6)%in the sham operation group(P<0.05).Conclusions Cell apoptosis occurred in both acinar and islet in the model of AOP,and this may be the pathophysiological basis of endocnne hormones alterations in the model of AOP.

BACKGROUND: In many studies, rats were commonly used as models of retrorsine-induced hepatic injury. Some reports have confirmed that retrorsine cannot inhibit proliferation of mouse hepatic cells. Other reports have shown that retrorsine has inhibitory effects on proliferation of mouse hepatic cells. OBJECTIVE: To study the liver regeneration after hepatic injury by creating mouse models treated with partial hepatectomy combination with retrorsine. METHODS: A total of 40 C57BL/6J mice were equally and randomly assigned to 2 groups. In the partial hepatectomy combined with retrorsine group, intraperitoneal injection of retrorsine 70 mg/kg was conducted, twice, within an interval of 2 weeks. Four weeks later, 2/3 hepatectomy was performed. In the partial hepatectomy group, intraperitoneal injection of saline 70 mg/kg was performed, twice, with an interval of 2 weeks. Four weeks later, 2/3 hepatectomy was performed. At 14 days after partial hepatectomy, the restoration of the livers was observed. The liver cell injury was observed at 3, 7 days with hematoxylin-eosin staining. The hepatocyte proliferation was observed at 3 days with BrdU staining. Oval cell proliferation was observed at 3, 7and 14 days with CK19 and C-kit antibody immunohistochemistry.RESULTS AND CONCLUSION: In the partial hepatectomy group, the damaged liver nearly restored to normal at 14 days after partial hepatectomy, and the result was contrary to partial hepatectomy combined with retrorsine group. Hematoxylin-eosin staining demonstrated that significant degeneration changes in hepatic cells in the partial hepatectomy combined with retrorsine group. BrdU staining showed that hepatocyte proliferation at day 3 was significantly determined in the partial hepatectomy group, but few in the partial hepatectomy combined with retrorsine group. CK19 and C-kit immunohistochemistry demonstrated that visible oval cell proliferation was seen in mice of partial hepatectomy combined with retrorsine group. First of all, hepatic oval cells appeared in portal area and differentiated into hepatic cells and bile duct cells, and then grew into the hepatic lobule gradually. These indicated that retrorsine can obviously inhibit hepatocyte regeneration after liver injury in mice. The model of mice treated with retrorsine and partial hepatectomy could induce oval cell proliferation.

Objective To investigate the effect of human bone marrow mesenchymal stem cells (MSCs) on proliferation of CD8+ T lymphocytes and its mechanism. Methods MSCs were isolated and cultured then identified through many ways. The proliferative influence of MSCs on peripheral blood mononuclear cells (PBMCs) stimulated by PHA was investigated. The effect of MSCs on proliferation of CD8 + T lymphocytes induced by PHA was explored by flow cytometry. The possible mechanism of the inhibition effect of MSCs was investigated on the proliferation of CD8+ T cells stimulated by PHA with Transwell assay. Results MSCs were successfully harvested and cultured in vitro. MSCs suppressed the proliferation of CD8+ T cells stimulated by PHA when MSCs ∶ PBMCs ≥ 1 ∶ 5, which showed a dose-dependent manner. Strong proliferative inhibition of MSCs was presented on the CD8 + T cells induced by PHA in the group of Transwell (MSCs ∶ PBMCs = 1 ∶ 1) and the influence was similar to non-Transwell group. Conclusion MSCs can affect body immunity via suppressing the proliferation of CD8+ T cells.

The treatment of lower extremity chronic refractory ulcers requires a long time but with poor prognosis. Thus, many patients end up with amputations. The treatment of lower extremity chronic and recalcitrant ulcers and limb salvage has been a challenge worldwide. The Ilizarov technique based on the law of "tension-stress" brings a new hope in treating lower limb chronic and recalcitrant ulcers. The Ilizarov technique and distraction osteogenesis not only induce bone formation but also lead to angiogenesis and improved microcirculation. The Ilizarov technique consists of longitudinal distraction of long bone and tibia trans-verse transport (TTT) (proximal tibial corticotomy followed by transverse distraction). These two techniques have their own advantages and disadvantages with different indications in clinical application. Longitudinal distraction of long bone is mainly used for bone formation, such as large bone defects, osteonecrosis or bone infection (with or without soft tissue loss or ulcers). Because of only a partial osteotomy in TTT, the trauma is minor and their effects on limb instability are limited. Moreover, the procedure is simple with only a few minor complications. Thus, it is ideal in treating lower limb ischemic ulcers, such as diabetic foot ulcers, thromboangiitis obliterans (Buerger's disease), ulcers caused by atherosclerotic occlusion, arterial or venous ulcers, and trauma wounds. Several studies reported that TTT achieved high healing and limb salvage rates in treating severe diabetic foot ulcer. However, TTT could achieve lower recurrent rate. Thus, it is the most successful application in treatment of chronic ulcers. TTT also improves healing and limb salvage in treatment of thromboangiitis obliterans. However, the overall effects are limited than those in treating diabetic foot ulcer. For lower limb atherosclerosis occlusion, TTT induces regeneration of microvessles and consequently leads to ulcer healing. The effects are better than other conventional treatments. A combination therapy with vascularization is emphasized to attain the optimal long-term effects. The effects of TTT on lower limb recalcitrant ulcers still need to be validated in randomized control trial with larger sample size. Further, the mechanism of treatment needs to be explored by pilot studies which could show that this may be related to the formation of pro-angiogenetic factors and a rebalance of the inflammatory microenvironment during TTT.

Objective By analyzing the perioperative management in our hospital to explore the clinical effect and safety of single kidney transplantation from deceased juveniles' donors.Methods We retrospectively analyze 86 cases of kidney transplantations from deceased juveniles' donors in our hospital from 2007 December to 2015 August.Results The success rate of the operations was 100％.The postoperative complications occurred as fellows:7 cases of acute rejection (8.14％);10 cases of drug intoxication (11.62％);21 cases of DGF (24.44％),4 cases of leakage of urine (4.65％),7 cases of lung infection (8.14％).Two cases (2.32％) died after the operation because of serious lung infection,and by corresponding treatment 47 cases recovered after 2-4 weeks.The creatinine level in 37 cases without any complications was 131.88 ± 44.20 μmol/L during discharge.Conclusion With strict selection,the organ from a deceased juvenile donor is safe and practicable.