The authors reviewed the research and development of tissue engineered meniscus from three points (cell seeds, scaffolds, cell factors) and point out current questions and investigative direction in the future.

BACKGROUND:Because of convenient source, multi-lineage differentiation and low immunogenicity, bone marrow mesenchymal stem cels are the ideal cel type to serve as vectors of transgenic cels in pain management. However, the replicative senescence and smal amount of cels obtained from the bone marrow restrict the application of bone marrow mesenchymal stem cels in pain research. OBJECTIVE:To construct human telomerase reverse transcriptase (hTERT)-immortalized rat bone marrow mesenchymal stem cels as transgenic celular vectors for pain therapy. METHODS:Bone marrow mesenchymal stem cels were obtained from whole rat bone marrow, and then transfected with a lentivirus containing the hTERT (pLV-Puro-EF1α-hTERT) folowed by puromycin selection. hTERT expression and telomerase activity in these transfected cels were determined by RT-PCR and TRAP. Morphological changes, capacity of cel growth and multi-lineage differentiation, chromosome karyotype and tumorigenicity were observed in vitro. Moreover, the expression of cel surface molecule, Nestin, MHC-I and MHC-II in transfected cels were also detected by flow cytometry and immunocytochemistry. RESULTS AND CONCLUSION: The bone marrow mesenchymal stem cels geneticaly modified by hTERT could be cultured and passaged through 30 generations in vitro. Compared to the primary and negative transfected cels, the hTERT-modified bone marrow mesenchymal stem cels showed higher expression of hTERT mRNA, telomerase activity and cel proliferation. Most of transfected cels stayed at G2/M and S stages. The proliferation index of the transfected cels were increased dramaticaly. The positive rates of CD29, CD44 and CD90 were over 70%, but the positive rates of CD34 and CD45 were less than 5%. Transfected cels were positive for Nestin in the cytoplasm, but negative for MHC-1 and MHC-11. In addition, this cel line continued to exhibit the characteristics of fibroblastic bone marrow mesenchymal stem cels, including phenotype, differentiation into osteoblasts, adipocytes and neuron-like cels. No chromosome abnormality and tumor formation were observed in this experiment. Taken together, these data suggests that the rat bone marrow mesenchymal stem cels immortalized by hTERT gene are constructed successfuly and stil maintain major stem cels characteristics, which provide safe and stable cel vectors as research base for pain therapy.

Objective To determine the potential impact of superantigens produced by skin-colonizing Staphyiococcus aureus in patients with atopic dermatitis and eczema. Methods Of 117 patients with atopic dermatitis and 199 with eczema, 140 Staphyiococcus aureus strains were isolated from the skin specimens. Superantigens were detected with reverse passive latex agglutination. Results Among 140 Staphyiococcus aureus strains, 60 (42.9%) produced superantigens, among which 43 produced one kind of superantigens only and 17 produced at least two kinds. Of strains isolated from atopic dermatitis, 51.5% produced superantigens and no significant difference was seen in superantigen production between lesional and non-lesional strains in atopic dermatitis. Of strains isolated from eczema patients, 34.7% (all were lesional strains) produced superantigens. The positive rates of total superantigens, lesional superantigens and toxic shock syndrome toxin-1 production were all higher in the strains from atopic dermatitis than in those from eczema. Conclusions Superantigen production by skin-colonizing Staphyiococcus aureus probably plays a more important role in atopic dermatitis than that in eczema. However, further studies are necessary to validate its importance.

Objective To evaluate the efficacy and advantages of the technique by combined PCNL and retrograde intrarenal surgery (RIRS) in a second stage to treat the complex renal stones in solitary kidney cases.Methods PCNL most with a single 18-24 F tract was performed first and RIRS was carried out at a second stage in solitary kidneys of 21 patients,of which congenital in 14.3％ (3 cases),contralateral nephrectomy in 42.8％ (9 cases),and functional solitry kidneys in 42.8％ (9 cases).Of the 21 patients,the average age was 45 years with 15 males and 6 females.The stones were 8 multiple,6 partial staghorn,and 7 complete staghorn with a mean size of 4.6 (3.8-6.8) cm.Results Of the 21 PCNL cases,a single tract,double,triple tracts were established in 18 (85.7％),2 (9.5％),1 (4.8％) cases,respectively.Mean operation time of PCNL was 95 (45-175) min.After 1 day of PCNL,all case had residual stones with a mean size of 1.9 (1.0-3.5) cm.Two case occurred fever after PCNL and one case presented bleeding resolved by selective renal artery embolization.The mean operation time of RIRS was 72 (35-95) min.Stone-free rate after RIRS was 85.7％ (18/21).The final stone free rate increased to 95.2％ (20/21) after one case received a second-look PCNL and two cases accepted ESWL.Two cases occurred fever and steinstrasses after RIRS resolved by rigid ureteroscopy.At the 3 months follow-up,renal function became stable,improved and worse in 71.4％ (n=15),23.8％ (n=5),and 4.8％ (n=l) of patients.Conclusions PCNL combined with RIRS could be an effective and safe option for complex stones in solitary kidneys with less bleeding,reduced tracts,minor complications and good renal function preservation.

Objective To compare the efficiency between mini-percutaneous nephrolithotomy (MPCNL) and extracorporeal shock wave lithotripsy (ESWL) for monotherapy of renal calculi in infants less than 3 years.Methods Forty-six infants were treated with either SWL (22 infants) or MPCNL monotherapy (24 infants).The mean age was (22.6 ± 8.7) months vs (23.5 ± 6.6) months and the stone size was (21.4 ± 3.5) mm vs (21.7 ± 1.7) mm,and there were no significant difference.Results For MPCNL,mean operating time was (76.2 ± 23.4) min and mean hospital stay was (14.13 ± 5.8) d.The stone-free rates were 84.0％ (21/25) after first session and 96.0％ (24/25) after second-look procedure.Postoperative fever happened in 4 (16.0％) cases.Hemoglobin drop was (8.5 ± 4.4) g/L and no one needed blood transfusion.For ESWL,the stone-free rate were 31.8％ (7/22) after first session and 86.3％ (19/22) after second session for 11 infants (50.0％).Mean hospital stay was (6.6 ± 2.3) d and 10 cases (45.5％)had complications.Hemoglobin drop was (10.6 ± 12.7) g/L.MPCNL was lower than ESWL in complications rate and re-treatment rate,and the stone-free rate was higher,but the hospital stay and operation time was longer (P ＜ 0.05).GFR revealed improve or stable after operation in both groups.Conclusions For a higher success rate,lower complication rate and re-treatment rate,MPCNL was an effective option for the management of relatively larger stones in children (even in infants).

The impact factors were explored to determine the horizontal positional relationship between the umbilicus and the 2nd lumbar spinal process in adults and to verify the accuracy of the localization of Shenshu (BL 23) via the umbilicus. The position of the umbilicus and the 2nd lumbar spinal process was measured in 100 participants and the data were analyzed through SPSS 20.0 software. It was found that the umbilicus and the 2nd lumbar process were not positioned horizontally. The positional relationship of these two sites was not apparently correlated with gender, age, body weight, body height, BMI, waistline and discomfort of lumbar region. The umbilicus was commonly and posteriorly projected on the site between the 4th and 5th lumbar vertebra. It is explained that the localization of Shenshu (BL23) via the umbilicus is not accurate.
Acupuncture Points ; Adolescent ; Adult ; Female ; Humans ; Lumbosacral Region ; anatomy & histology ; Male ; Meridians ; Middle Aged ; Umbilicus ; anatomy & histology ; Young Adult

Objective To establish a method for isolating and culturing fibroblasts from cirrhotic liver in vitro,observe the biological characteristics of fibroblasts.Methods Human fibroblasts from cirrhotic liver were isolated and cultured by adhering to the culture plastic.The morphologic and growth characteristics of the acquired cells were observed by microscopy imaging and cell counting.The expression of FSP-1 and Vimentin of fibroblasts was detected through immunofluorescence and western blotting.Results The tissue blocks were well-adherent to the culture plastic within 2 hours.Hepatocyte-like cells were observed surrounding the blocks within about 1 week,and spindle-shaped fibroblasts were observed within about 2 weeks.After a series of passage,the attached cells became proliferated quickly.The growth curves of the passage 3,4 and 5 were quite similar.The result of immunofluorescence showed the expression of FSP1 and Vimentin was positive.The positive rate was respectively (90.6 ± 1.0) ％ and (91.2-± 4.1) ％.Conclusions Human fibroblasts from cirrhotic liver can be cultured successfully through adherent property.The method for isolation and culture of fibroblasts from cirrhotic liver is convenient,efficient,stable and cultured cells remain naive biological characteristics.

ObjectiveTo investigate the clinical value of fluorescence in situ hybridization (FISH) in the diagnosis of urothelial carcinoma.MethodsFrom September 2010 to October 2011,morning voiding urine of 27 patients with suspected urothelial carcinoma was collected for FISH examination.The results of FISH examination were compared with the results of pathological examination by endoscopic biopsy.Both sensitivity and specificity were compared respectively,and the cost of each kind of examination was also considered.ResultsOf 27 cases,pathological examination by endoscopic biopsy revealed 11 cases of urothelial carcinoma,FISH examination showed 9 cases of urothelial carcinoma,but only 7 cases in these 11 cases.The overall sensitivity of FISH examination was 63.6％ (7/11 ),the specificity was 87.5％(14/16).The cost per case of FISH examination (3100 yuan) was 3.1 times of pathological examination by endoscopic biopsy( 1000 yuan).ConclusionsIt showed that there is no advantage of FISH examination for diagnosis of urothelial carcinoma considering both the sensitivity and specificity,and the cost is also higher than that of pathological examination by endoscopic biopsy.It should be evaluated further when FISH examination is widely used in clinic.

Objective To investigate the effects of high-fat diet on pancreatic acinar cells' IP3 expression and CCK-induced amylase release in rats.Methods Male Wistar rats were divided into high-fat diet group and normal diet group,they were fed for 4 weeks.Blood triglycerides,cholesterol,amylase and glucose levels were determined by automatic biochemical analyzer.Pancreatic tissues were taken for histopathological observations.Pancreatic acinar cells were isolated and cultured,and intracellular inositol 1,4,5-trisphosphate (IP3) was detected using a commercial kit.Amylase release rates were measured after CCK-8 stimulation.Results The rats in high-fat diet group appeared hyperlipidemia,vacuolization of acinar cells and the lymphocytes appeared around the acinar cells can be seen on the pancreatic tissue pathology staining.The levels of IP3 in acinar cells of rats fed a high-fat diet were higher than that of normal rats [(31.807 ± 3.448) pmol/106 cells vs (24.632 ± 3.649) pmol/106 cells,t=7.479,P＜0.001];and amylase release rate in these rats'acinar cells were also higher than those of normal rats [when CCK-8=0.01 nmol/L:( 11.056 ±3.369)％ vs (7.354 ± 2.181) ％,t=3.912,P＜0.001;when CCK-8=1 nmol/L:( 13.854 ± 4.087 ) ％ vs (9.432 ±2.477) ％,t=3.939,P＜0.001 ) after CCK-8 stimulation in different concentrations.Additionally,there was a positive co-relationship between acinar cell's IP3 level and amylase release (r=0.896,P＜0.001 ).Conclusion Chronic high-fat diet induces hypersensitivity for pancreatic acinar cells' exocrine function,and IP3 as a signal molecule may play an important role in this process.

Aim To investigate the role of endothe-lium in the enhancement of phenylephrine-mediated vasoconstriction by bupivacaine in the isolated rat aor-ta.Methods The isolated rat aortic rings were pre-pared, and the vascular endothelium was removed chemically or physically .Phenylephrine-mediated vas-oconstriction was recorded .Results A pretreatment with bupivacaine at 30 μmol · L-1 for 20 min signifi-cantly increased the Emax value of vasoconstrictive re-sponses to phenylephrine from 2.22 ±0.07 g of sol-vent-controlled group to 2.50 ±0.05 g ( P<0.01 ) in the isolated endothelium-intact rat aorta.However, the Emax value was not significantly changed by a pretreat-ment with bupivacaine at 30 μmol · L-1 for 5 , 10 or 15 min ( P>0.05 ) .A pretreatment with bupivacaine at 30 μmol · L-1 for 20 min slightly but significantly inhibited the vasoconstrictive responses to low concen-tration of phenylephrine in the isolated endothelium-de-nuded rat aorta (P<0.05).In the isolated endotheli-um-intact rat aorta under a combined treatment with in-dometacin, ChTX, apamin and L-NAME, the vasodi-lator responses to acetylcholine were completely sup-pressed , and a pretreatment with bupivacaine at 30μmol· L-1 for 20 min did not significantly affect the vasoconstrictive responses to phenylephrine ( P >0.05 ) .Conclusion Bupivacaine enhances α1-adre-noceptor-mediated vasoconstriction by inhibiting vascu-lar endothelium in the isolated endothelium-intact rat aorta, Which potentiates indirectly the vasoconstrictive responses to phenylephrine .