BACKGROUND: Proliferation and differentiation of mesenchymal stem cells (MSCs) is associated with platelet concentration in platelet-rich plasma (PRP). Low enrich multiple cannot reach proper effects, but high level had inhibitory effects on osteoanagenesis.OBJECTIVE: To observe the effect of different volume fraction of PRP on dog BMSC proliferation.DESIGN: A cytological in vitro study.MATERIALS: Healthy 12-month male Beagle dogs were supplied by the Experimental Animal Center, Xiangya Medical College,Central South University.METHODS: Dog BMSCs of 5 passage were adjusted to 3×10~8/L, and incubated in a 96-well plate at 200 μL per well. Following 24 hours of routine culture, primary medium and non-adherent cells were discarded. Prepared PRP gel was mixed with serum-free low-glucose DMEM containing penicillin and streptomycin, and then diluted into 5%, 6.25%, 7.5%, 8.75%, 10% volume fraction. 200 μL above-described liquid was added into the 96-well plate, which was subsequently placed in a incubator.We set up a blank control.MAIN OUTCOME MEASURES: MTT was used to investigate effect of different volume fraction of PRP on dog BMSC proliferation.RESULTS: Compared with the blank control group, various volume fraction of PLP could promote dog BMSC proliferation in early stage. With prolonged time, proliferation speed began to increase at day 6 in the 8.75% and 10% PRP groups, entering platform stage. BMSC number was increased rapidly in the 5% and 6.25% PRP groups, especially in the 6.25% PRP group.CONCLUSION: PRP gel could promote BMSC proliferation markedly and proliferation strength of BMSCs was correlated to the density of PRP. BMSC proliferation would be accelerated by the low density of PRP.

BACKGROUND: Platelet-rich plasma (PRP) contains abundant growth factors that were needed for osteanagenesis. Moreover,the proportion of each growth factor formed by an organism, with good synergism.OBJECTIVE: To explore the influence of PRP on osteogenetic activity of canine bone marrow mesenchymal stem cells (BMSCs) after induction in vitro.DESIGN, TIME AND SETTING: The in vitro cytological experiment was performed at the Central Laboratory of Xiangya Hospital of Central South University from June 2007 to February 2008.MATERIALS: Healthy 12-month male Beagle dogs were supplied by the Experimental Animal Center, Xiangya Medical College,Central South University.METHODS: The 3~(rd) generation BMSCs were collected and divided into 4 groups. BMSCs in the control group were incubated in standard medium. BMSCs in the osteogenetic induction medium group were incubated in high-glucose DMEM containing fetal calf serum, dexamethasone, beta-sodium glycerophosphate and vitamin C. BMSCs in the PRP group were incubated in low-glucose DMEM containing 6.25% PRP. BMSCs in the combination group were incubated in high-glucose DMEM containing 6.25% PRP, dexamethasone, beta-sodium glycerophosphate, and vitamin C.MAIN OUTCOME MEASURES: Alkaline phosphatase activities were measured. Expression of collagen type I was examined by immunocytochemical staining. Calcium tuberoses were labeled using modified Von Kossa staining. Expression of osteocalcin mRNA was examined by RT-PCR.RESULTS: Levels of alkaline phosphates of all groups became increased along with time. The alkaline phosphates level of combination group was strongest (P < 0.05). Following 7 and 14 days of induction, type I collagen expressed positively in the osteogenetic induction medium and combination groups, but negatively in the PRP and control groups. Following 14 days,formation of calcium nodules were found in the osteogenetic induction medium and combination groups. Following 7 and 14 days,expression of osteocalcin mRNA were similar between the control and PRP groups (P > 0.05), which was significantly lower than the osteogenetic induction medium and combination groups (P < 0.05). Expression of osteocalcin mRNA was significantly lower in the osteogenetic induction medium group compared with the combination group (P < 0.05).CONCLUSION: PRP gel can effectively promote osteoblastic effect of BMSCs after induction in vitro following induction in osteogenetic medium.

Objective To study the the causes, management and preventive measures for complications of holmium laser ureterolithotripsy. Methods Three hundred and seven cases of uteteral calculi from Jan. 2006 to Jan. 2010 were reported (203 men and 104 women, age ranged from 18-83 years,mean age 51.6 years). One hundred thirty-three patients had calculi in the lower ureter,94 in the middle part and 80in the upper ureter. Results The overall comminution rate was 89.9% (276/307 cases). sixteen cases (5.2%) were followed by ESWL due to the shift of the stones to the renal pelvis. The operation failed in 9 cases(2.9%), 3 cases(1%) had perforation,and recoveried after open operation. Deep infection was found in 3(1%)cases,which recoveried after anti-infective therapy. Conclusions Holmium laser ureterolithotripsy is an effective method for treating ureteral calculi, with advantage of high efficiacy,safety and minimal invasion. To master the operating induction, to raise operating technique and to take correct measure could reduce complications effectually.

OBJECTIVETo observe the efficacy and safety of Yigu capsule (YGC, a Chinese herbal compound preparation) in treating postmenopausal osteoporosis (PMO) and to explore its possible mechanism.

METHODSThe clinical study was conducted adopting prospective, randomized, double blinded method for 6 months with placebo and positive controls. Two hundred and ten PMO patients with confirmed diagnosis were divided into the YGC group, the osteocalcin group and the placebo group, they were treated with YGC, osteocalcin capsule and placebo capsule, respectively. The symptoms, as new fracture and ostealgia, bone mineral density (BMD) of the 2nd to the 4th lumbar vertebrae (L24) and upper segment of femur, blood and urinary indexes for bone metabolism, sex hormone level and adverse reaction were observed.

RESULTSIn the YGC group, the total effective rate was 95.50%, no new fracture occurred, which was significantly better than that in the other two groups (P < 0.05). The increase of BMD was 9.83% in L2-4, 4.09% in femoral neck, 4.60% in Wards triangle, 3.00% in greater trochanter, which were better than those in the placebo group (P < 0.05, P < 0.01). As compared with the placebo group, in the YGC group, levels of urinary oxyproline hydroxyproline/creatinine, urinary calcium/creatinine were lower, serum and bone alkaline phosphatase, osteocalcin, estradiol, and estradiol/testosterone were higher, but with no difference in level of testosterone. In the observation period, no abnormal findings in blood and urine routine examination as well as in liver and renal function were found. Mild, transient gastro-intestinal response occurred in individual patients but it didn't affect the treatment.

CONCLUSIONYGC could treat PMO effectively, it could obviously increase the BMD of lumbar vertebrae and hip, elevate the alleviating rate of ostealgia and incessant motion time, without new compressive fracture of vertebrae, and without any related adverse reaction. YGC could not only promote the formation, but also inhibit the absorption of bone, and increase the sex hormone level, therefore, it is a pure Chinese herbal compound preparation that worths further deep research and development.

Absorptiometry, Photon ; Bone Density ; Capsules ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Estradiol ; blood ; Female ; Humans ; Middle Aged ; Osteoporosis, Postmenopausal ; drug therapy ; physiopathology ; Phytotherapy ; Prospective Studies ; Testosterone ; blood

PURPOSE: Because previous studies have reported depleted antioxidant capacity in patients with chronic pancreatitis (CP), prevention of free radical production has gained importance in antifibrotic treatment strategies for CP. The aim of this study was to investigate the effects of ascorbic acid on oxidative capacity and pancreatic damage in experimental CP. MATERIALS AND METHODS: CP was induced in male Sprague-Dawley rats by infusion of dibutyltin dichloride (DBTC) into the tail vein. Ascorbic acid was given intraperitoneally at a daily dose of 10mg/kg body weight. The treatment groups were as follows: group 1, DBTC plus intraperitoneal physiologic saline; group 2, DBTC plus intraperitoneal ascorbic acid; group 3, solvent plus intraperitoneal physiologic saline; group 4, no operation plus intraperitoneal physiologic saline. Each group contained 15 animals. Treatment was started after CP was established. After 4 weeks of treatment, serum hyaluronic acid and laminin levels were determined by radioimmunoassay, pancreatic tissue oxidative stress was analyzed, and the degree of pancreatic damage was determined. RESULTS: Ascorbic acid treatment markedly increased superoxide dismutase (SOD) activity and decreased malondialdehyde (MDA) concentrations in pancreatic tissue (p < 0.01 for both). Significant serum hyaluronic acid and laminin reductions were observed in group 2 as compared with group 1 (p < 0.05). However, the serum hyaluronic acid and laminin levels remained elevated when compared with those of groups 3 and 4 (p < 0.05). Histopathologic scores were also lower in animals with CP that underwent ascorbic acid-treatment (p < 0.05). CONCLUSION: Ascorbic acid treatment alleviated the degree of oxidative stress and pancreatic damage in rat CP. Antioxidant treatment might be considered a potential option to improve the pathologic process in CP.
Animals ; Antioxidants/pharmacology ; Ascorbic Acid/*pharmacology ; Hyaluronic Acid/blood ; Laminin/blood ; Male ; Organotin Compounds ; Oxidative Stress/drug effects ; Pancreas/*drug effects/pathology ; Pancreatic Diseases/blood/chemically induced/*prevention & control ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the mechanism of acupoint sticking of "Hua yutie" in improving ischemic stroke.

METHODSEighty rats were randomly divided into 5 groups, a model group, an acupoint sticking group, an acupuncture group, a Nimodipine group and a normal group. Middle cerebral artery occlusion (MCAO) was used for preparation of focal cerebral ischemic rat model. After modeling, any treatment was not given to the model group; for the acupoint sticking group, "Hua yutie" was applied at "Dazhui" (GV 14) ,"Qihai" (CV 6) and "Mingmen" (GV 4); for the acupuncture group, acupuncture was given at the same acupoints as those in the acupoint sticking group; the Nimodipine group received intragastric administration of Nimodipine. And the normal group did not receive any treatment. Their infarction volume, the cerebral water content, expression of vascular endothelial growth factor (VEGF) and the protein level were observed.

RESULTSThe infarction volume coincided with the dominative scope of the middle cerebral artery of the electric coagulation. There were significant differences in the cerebral water content as the various treatment groups compared with that of the model group (all P<0.05). The VEGF positive cell number and the protein level around the infarction area in the acupoint sticking group were increased as compared with those in the model group (P<0.01), with no significant difference as compared with the Nimodipine group and the acupuncture group (all P>0.05).

CONCLUSIONAcupoint sticking of "Hua yutie" alleviates the cerebral damage after ischemia possibly through enhancing the expression and protein level of VEGF.

Acupuncture Points ; Administration, Cutaneous ; Animals ; Brain Ischemia ; drug therapy ; genetics ; metabolism ; Cerebral Infarction ; drug therapy ; genetics ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Gene Expression ; drug effects ; Humans ; Male ; Rats ; Rats, Wistar ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo evaluate the available surgical treatment modalities so as to explore the optimal strategy of managing early breast cancer.

METHODSThe clinical data of 2173 consecutive early-stage breast cancer patients treated by surgery treatments were retrospectively reviewed in order to clarify the indications and contraindications of different modalities. Therapeutic outcome of different surgical treatment modes were compared in terms of recurrence-free survival ( RFS) , disease-free survival ( DFS) , overall survival (OS). The cosmetic results of breast conservation and reconstruction were also evaluated .

RESULTSThe median age of these patients was 51 years ranging from 18 to 91. Of 2173 patients, 547 had stage 0- I lesions and 1626 stage II , and 1155 (53. 2% ) premenopausal. The proportion of patients who received radical surgery, breast conservation and reconstruction after mastectomy was 83. 6% (1817/2173), 10. 5% (229/2173) and 2. 5% (55/2173) , respectively. Younger and premenopausal patients prefer conservative and reconstructive surgeries, which are reasonable for stage 0-I and non-invasive breast cancer patients. Conservative surgery was not suitable for Paget's disease of breast (P = 0. 004) , mastectomy followed by reconstruction in this type of cancer was up to 38. 5%. The recurrence and metastasis rate of conservation or mastectomy were similar with a comparable 3-year RFS of 97. 4% and 95. 4% , respectively; there were also no significant differences in RFS(P =0. 2435) , DFS( P =0. 1395) and OS(P =0. 9406) after having been followed for 3 to 64 months. Similarly, immediate reconstruction did not show any negative effects with only 1 recurrence and 1 metastasis. Aesthetic outcomes were assessed as excellent or good in 90. 0% of breast conservation surgery, and the acceptability of reconstruction was 94. 5%.

CONCLUSIONBreast conserving surgery not only has comparable survival as mastectomy, but also has better cosmetic outcomes. Immediate breast reconstruction can be a suitable option without compromising survival. It is very important in the management for early breast cancer by selecting the most suitable surgery mode for every individual patient not only to cure her disease but also to satisfy the patient psychologically. Conservation should be preferred prior to reconstruction whenever possible.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; pathology ; surgery ; Carcinoma, Ductal, Breast ; pathology ; surgery ; Carcinoma, Intraductal, Noninfiltrating ; pathology ; surgery ; Disease-Free Survival ; Female ; Follow-Up Studies ; Humans ; Lymphatic Metastasis ; Mastectomy ; methods ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Paget's Disease, Mammary ; pathology ; surgery ; Reconstructive Surgical Procedures ; Retrospective Studies

OBJECTIVETo evaluate the application of fluorescence in-situ hybridization (FISH) in detection of gene translocation in paraffin-embedded tissue samples of synovial sarcoma.

METHODSInterphase FISH was carried out in paraffin-embedded tissue of 42 cases of synovial sarcoma and 9 cases of non-synovial sarcoma, using a LSI SYT (18q11.2) dual color break-apart probe. In all of the cases studied, the gene fusion product SYT-SSX was also analyzed by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSPositive signals were detected in 37 cases (88.1%) of synovial sarcoma by FISH, as compared with 35 cases (83.8%) by RT-PCR and 39 cases (92.9%) by both techniques. Of the 39 positive cases, 33 cases (78.5%) revealed SYT gene translocation.

CONCLUSIONSFISH may serve as an adjunctive diagnostic tool in problematic cases of synovial sarcoma and can be applied in paraffin-embedded tissue samples. As compared with RT-PCR, FISH is also sensitive and reliable. The methodology is less labor intensive and time consuming. FISH has great potential in molecular diagnosis of soft tissue tumors.

Adolescent ; Adult ; Biomarkers, Tumor ; genetics ; Child ; Child, Preschool ; Chromosome Aberrations ; Female ; Head and Neck Neoplasms ; genetics ; metabolism ; Humans ; In Situ Hybridization, Fluorescence ; Lower Extremity ; pathology ; Male ; Middle Aged ; Oncogene Proteins, Fusion ; genetics ; Paraffin Embedding ; Reverse Transcriptase Polymerase Chain Reaction ; Sarcoma, Synovial ; genetics ; metabolism ; Soft Tissue Neoplasms ; genetics ; metabolism ; Young Adult

Objective Normal nucleus pulposus cells (nNPCs) can induce the differentiation of mesenchymal stem cells (MSCs).However, the inductive effect of degeneration nucleus pul-posus cells ( dNPCs ) on MSCs has not been determined .This study aimed to compare nNPCs with dNPCs in inducing MSCs into nucleus pulposuslike cells. Methods MSCs (cell line) were co-cultured with NPCs (cell line/isolated from patients with intervertebral disc degeneration) in 3D in vitro.The cells were divided into 5 groups:nNPCs control group, dNPCs control group, MSCs control group, nNPCs-MSCs group (MSCs induced by nNPCs), dNPCs-MSCs group (MSCs induced by dNPCs).After co-cultured for 7 days, the expression level of aggrecan (ACAN), type II collagen (COL-2), SOX-9, matrix metalloproteinase-1 (MMP-1) and MMP-3 in all the 5 groups'cellswere detected by RT-PCR and Western-blot. Results The RT-PCR results showed the expression of ACAN , COL-2 and SOX-9 in nNPCs-MSCs group was higher than that in dNPCs control group and dNPCs -MSCs group (all P<0.05).The expression of MMP-1 and MMP-3 in nNPCs-MSCs group and dNPCs-MSCs group was lower than that in dNPCs control group ( all P<0.01) , but there was no difference when compared with nNPCs control group and MSCs control group (all P>0.05).Western-blot analysis showed that the expression of ACAN , COL-2 and SOX-9 in nNPCs-MSCs group was higher than that in dNPCs control group and dNPCs-MSCs group (all P<0.05).The expression of MMP-1 and MMP-3 in nNPCs-MSCs group and dNPCs-MSCs group was lower than that in nNPCs control group and dNPCs control group ( all P<0.01) , but there was no difference compared with MSCs control group ( all P>0.05) . Conclusion Both nNPCs and dNPCs could induce the differentiation of MSCs into nucleus pulposus cells .After co-cultured with nNPCs, the expression level of ACAN, COL-2 and SOX-9 of MSCs was similar to that of nNPCs, which was superior to the induc-tion of MSCs by dNPCs under the same culture conditions .

OBJECTIVETo observe the efficacy and safety of Yigu capsule (YGC), a Chinese herbal compound preparation, in treating postmenopausal osteoporosis (PMO) and to explore its possible mechanism.

METHODSThe clinical study was conducted in a prospective, randomized, double blinded method lasting for 6 months with placebo and positive control. Two hundred and ten PMO patients with confirmed diagnosis were assigned into the YGC group, the calciferol group and the placebo group. Besides being administered element calcium, they were treated with YGC, calciferol capsule and placebo capsule respectively. And such symptoms as newly found fracture and ostealgia, bone mineral density (BMD) of the 2nd-4th lumbar vertebrae (L(2-4)) and upper femur, blood and urinary indexes for bone metabolism, sex hormone level and adverse reaction that occurred in patients were observed.

RESULTSIn the YGC group, the total effective rate was 95.50%, with no new occurrence of fractures, which was significantly better than that in the other two groups (P < 0.05). Moreover, in the YGC group, the increase rate of BMD was 9.83% in L(2-4), 4.09% in femoral neck, 4.60% in Wards triangle, 3.00% in greater trochanter, which was also better than that in the placebo group (P < 0.05, P < 0.01). As compared with the placebo group, levels in the YGC group of urinary oxyproline hydroxyproline/creatinine, urinary calcium/creatinine were significantly lower, serum and bone alkaline phosphatase, osteocalcin, estradiol and estradiol/testosterone were significantly higher, but no difference was shown in the comparison of testosterone level. In the observation period, no abnormality in blood or urine routine, liver or renal function was found. Only mild, transient gastro-intestinal response occurred in individual patients, but it did not affect the treatment.

CONCLUSIONYGC could treat PMO effectively, as it could obviously increase the BMD of lumbar vertebrae and coxafemoral bone, elevate the alleviating rate of ostealgia and incessant motion time, yet causing no newly found compressive fracture of vertebrae, or and any related adverse reaction. YGC could not only promote the formation, but also inhibit the absorption of bone as well as increase the sex hormone level. Therefore, it is a pure Chinese herbal compound preparation worthy of further research and development.

Administration, Oral ; Aged ; Amidohydrolases ; urine ; Bone Density ; drug effects ; Bone and Bones ; drug effects ; metabolism ; Calcium ; administration & dosage ; blood ; urine ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Female ; Fractures, Bone ; epidemiology ; prevention & control ; Gonadal Steroid Hormones ; blood ; Humans ; Hydroxyproline ; urine ; Incidence ; Middle Aged ; Osteoporosis, Postmenopausal ; drug therapy ; metabolism ; Prospective Studies ; Treatment Outcome