Objective Get a primary culture method of osteoblast which is simple to operate and has many pure cells. Method Get osteoblast with improved I collagenase, and test the form of osteoblast, the alkaline phosphatase dyeing and formation of mineralization node. Result The improved rats primary culture method of osteoblast can get most cells in normal shape, alkaline phosphatase secretion and mineralization. Conclusion The improved rats primary culture method of osteoblast is convenient to operate, and can get most cells.

ObjectiveTo sum up the experience on diagnosis and treatment for patients of hyperthyroidism complicated by thyroid cancer. MethodsBetween 1994 and 2000, 300 hyperthyroidism patients were admitted, among them 12 suffered concomittent thyroid carcinoma and received surgical therapy and pathologic examination. ResultsIn this series thyroid carcinoma occurred in 4%(12/300) in patients with hyperthyroidism, but correct preoperative diagnosis was obtained in only one (3%) of the 300 cases. Bilateral subtotal thyroidectomy was initially performed in all 12 cases with 7 cases undergoing additionally ipsilateral total thyroidectomy when a cancer was identified. Two cases underwent functional cervical clearance of lymph nodes. Postoperative follow-up of 9 mos to 6 years found no recurrence. ConclusionIn patients of hyperthyroidism a thyroid cancer may develop on the background of immunodeficiency and endocrine dysfunction. In patients with hyperthyroidism, a thyroid nodule found perioperatively must be biopsied for frozen section to guide the surgical modality.

Expression of endogenous ouabain in placenta and the concentrations of serum ET-1 and NO were examined in 30 patients with hypertensive disorder complicating pregnancy (HDCP) and 30 healthy pregnant women to investigate the effect of endogenous ouabain on HDCP. Compared with the healthy pregnant group, the expression of endogenous ouabain dramatically increased in the HDCP groups (P<0.01). There was a significantly positive correlation between the expression of endogenous ouabain with ET-1 (r=0.5567, P<0.01), while the correlation of endogenous ouabain and NO was significantly negative (r=-0.6895, P<0.01). As expected, the correlation between ET-1 and NO was negative (r=-0.7796, P<0.01). ET-1 concentrations of maternal and cord sera in HDCP groups were significantly higher in comparison with healthy pregnant group (P<0.01). On the contrast, NO concentrations were much lower in the maternal and cord sera of HDCP groups as compared with healthy pregnant group (P<0.01). Our data suggest that endogenous ouabain is directly involved in the nosogenesis of HDCP, with accompanying decreased NO and the elevated of ET-1.
Case-Control Studies ; Endothelin-1/*blood ; Hypertension, Pregnancy-Induced/*metabolism ; Nitric Oxide/*blood ; Ouabain/*metabolism ; Placenta/*metabolism

[Objective] To investigate the difference of bone Absorption function of the rat osteoclast(OC) in different cultural methods, and the difference in the levels of mRNA expression of bone Absorption key gene, ATPase a3 gene, and provide the basis for further investigation on the OC in vitro. [Methods] with mechanical separation method, mature osteoclast was separated from inner wall of 24 hours newborn rat long bones medullary cavity; and induction culture methods, bone marrow cell was induced to osteoclast like cell(OLC) by using 1,25(OH)2D3.The morphological and functional change of osteoclast was observed and the difference in the level of ATPase a3 mRNA expression was determined.[ Result]OC and OLC are multinucleated giant cells, which can be stained positive by potartrate resistant acid phosphatase(TRAP), and can form bone Absorption lacuna. The number of OLC in induction method is more than in mechanical separation method, but bone Absorption lacuna is smaller and shallower than the early stage of induction. The late stage of OCL is extremely similar to OC. There is no significant difference of ATPase a3 mRNA expression from the cells between mechanical separation 8 hour and induction culture 6 days, but either of them is significantly less than inducing 8 days. [Conclusion]Induction method can produce a large number of OLC which is superior to mechanical separation method, but its bone resorption function is weaker in the earlier stage, because bone resorption function is associated with the number of nucleus. The late stage of OCL is extremely similar to OC of mechanical separation method. And it can be used in any experiments.

Objective To compare the therapeutic effects between cervical and extra-cervical surgical approaches for endoscopic thyroidectomy . Methods From October 2012 to December 2013, forty-four thyroid goiter patients were divided into two groups randomly .Group A underwent modified Miccoli endoscopic thyroidectomy ( n =20 ) and group B underwent endoscopic thyroidectomy via breast areola approach ( n=24 ) .The operative time , intraoperative blood loss , pain in 24 hours postoperatively , drainage volume , postoperative hospital stay , hospital cost , complication and cosmetic results between the two groups were compared . Results Compared with group B, group A had shorter operative time [(77.9 ±28.3) min vs.(97.9 ±30.0) min, t=-2.259, P=0.029], less intraoperative blood loss [(15.9 ±8.7) ml vs.(29.5 ±16.1) ml, t=-3.384, P=0.002], less pain in 24 hours postoperatively ( no pain, endurable pain , intolerable pain in group A and B were 15, 5, 0 and 7, 15, 2 cases respectively , Z=-3.066, P=0.002), less postoperative drainage volume [(31.7 ±10.3) ml vs.(57.0 ±14.6) ml, t=-6.511, P=0.000], but poorer cosmetic results (very dissatisfied, not satisfied, satisfied, comparatively satisfied, and very satisfied in group A and B were 1, 4, 5, 6, 4 and 0, 1, 4, 5, 14 cases respectively, Z=-2.723, P=0.006).There was no significant difference in postoperative hospital stay and hospital cost between the two groups (P>0.05).One case suffered transient hoarseness in group A and one case had trembling hand due to low calcium level in group B and both of them recovered 1 month after symptomatic treatment .No permanent recurrent laryngeal nerve injury , parathyroid injury or other complications occurred in both groups . Conclusions Cervical approach is minimally invasive and leads to good cosmetic results while extra-cervical approach causes bigger trauma but leads to better cosmetic results.Patients with high cosmetic reguest may choose endoscopic thyroidectomy via breast areola approach .

Objective To identify significantly differentially expression of rat by genes chip,try to find out the initiating factors and molecular mechanisms that oxidative stress originate or strengthen the steriod-induced necrosis of femoral head(SINFH).Methods Twenty Wistar rats were dived into experimental group and control group randomly.The rats were injected intraperitoneally whith endotoxin,and then injected intramuscularly with high-dose methylprednisolone or saline in experimental group and control group respectively.The mRNA was extracted from the femoral head of rats inevery group,and the cDNA probes were obtained by inverse transcript,and then carried out microarray detection.The quantitative RT-PCR was used to confirm the results of the microarray.Results Histopathological findings revealed that the experimental group rats had femoral head necrosis,trabecular bone disorders,thinning,bone cell necrosis,and the rate of empty lacunae increased,and in control group no femoral head osteonecrosis was found.Total of 27 differentially expressed genes were found,and of which 4 genes(COX6A2,COX4I2,SOD3,and DUSP1)were significantly different.The expression of these 4 genes were down-regulated.The functions of these four genes involved in inhibition of reactive oxygen species(ROS)generation,accelerated removal of ROS and protection tissue from oxidative damage and so on.Conclusion The expression of oxidative stress-related genes in SINFH of rats exist change.COX6A2,COX4I2,SOD3,and DUSP1 are key genes in process of oxidative stress originate or strengthen the SINFH.

Objective To investigate the feasibility and safety of helix hydro-jet lithotrity in treatment of calculus of intrahepatic duct and to establish its basic parameters.Methods The parameter of hydro-jet Was set at different levels of pressure(5000-9000 kPa)and different nozzle directions,and the effect of different parameter wag observed and analyzed in three groups.including 32 ex vivo intrahapatic gallstones.thirty-two ex vivo swine bile ducts and 31 ex vivo bile ducts obtained from patients with hepatolithiasis.Results Lithotrity was successful in all groups with a jet pressure of 5000-9000 kPa.The multiple regression analysis showed that the time of procedure depended linearly on the minimal stone diameter,stone weight andjet pressure.The time of procedure and the time needed to damage the bile duct decreased with the increase in jet pressure.At same jet pressure,the smaller the angle between nozzle and bile duct,the longer the time needed to cause damage in bile duct.The technique was successfully performed in 60 patients,with no severe complications,including bile duct injury.Conclusion It is feasible to treat hepatolithiasis with hydro-jet lithotrity.The most optimal operation condition was hydro-jet pressure of 6000-7500 kPa and frequency of 1 Hz.

Objective To investigate the formation and special cell biological behavior of osteoclasts.Methods The live-cell imaging technology was adopted to consecutively and dynamically observe the whole process of multinuclear osteoclast formation induced by RANKL and M-CSF from rat peripheral blood monocyte.Meanwhile,the inverted phase contrast microscopy,TRAP staining,and scanning electron microscopy were also applied to identify the osteoclast.Results After 2-week cultivation,a great number of apocytes were found by the inverted phase contrast microscopy,and most apocytes and monocytes had positive reaction after TRAP staining.Moreover,many bone resorption lacunae in which osteoclasts were perhrming bone resorption function could be found in the bone slice under the scanning electron microscope.Live-cell imaging observation showed that the multinuclear osteoclasts were generated through self-fusion of monocytes,fusion of monocytes and apocytes,as well as fusion between apocytes.All fusion processes occurred under the condition of cell adherence.Time-lapse Microcinematography observation showed diverse shapes of osteoclasts and the cell division of multinuclear osteoclasts.Conclusion Rat peripheral blood monocyte can develop into osteoclast under induction of RANKL and M-CSF.Osteoclast can form gigantic apocyte via various types of cell fusion to increase its nucleus number and cell volume,vary its shape,and increase the area of plasma membrane.On the other hand,osteoclast can decrease its cell volume and nucleus number via cell division to adapt the needs of local morphology,biomechanics and bone resorption dynamics.It suggests that this non-mitosis cell division is a special cell biological behavior of osteoclast,which may be the basis of exerting its function and improving bone resorption efficiency.

Objective To systematically review the efficacy and safety of Jinlian Qingre Effervescent Tablet (JQET) compared to Ribavirin for acute upper respiratory tract infection in children.Methods CNKJ,CBM,WanFang Data,VIP,PubMed,EM base,Web of science,and Cochrane Library databases were searched from the date of establishment to December 2016 for all randomized controlled trials (RCTs) and quasi-RCT on the use of JQET in children with acute respiratory infections.Meta-analysis by using Rev Man 5.3.Results A total of seven RCTs involving 782 patients were included,while the group of JQET involving 392 patients,Ribavirin involving 390 patients.The results ofmeta-analysis indicated that the efficacy in Jinlianqingre group was superior to that of Ribavirin control group,such as clinical effectiveness [RR=l.26,95％CI=(1.18,1.34),P＜0.000 01],fever subsidence time [MD=-1.54,95％CI (-1.79,-1.30),P＜0.000 01],the time of subsided cough [MD=-1.53,95％CI (-1.79,-1.27),P＜0.000 01],the disappearance time of pharyngalgia [MD=-1.29,95％CI (-1.88,-0.70),P＜0.000 1],and Pharyngeal congestion disappearance time [MD=-2.80,95％CI(-3.11,-2.49),P＜0.000 01].The difference was statistically significant.There were three adverse reactions reported in JQET group.Conclusion JQET is superior to the Ribavirin control group in clinical effectiveness,fever subsidence time,time of subsided cough,disappearance time of pharyngalgia,and pharyngeal congestion disappearance time to treat acute upper respiratory tract infection in children.However,these results should be carefully interpreted,and this conclusion has to be further verified by high quality,large scale RCTs.

Limulus Factor C, a serine protease zymogen from the amoebocytes of the limulus, has high affinity for endotoxin. When Factor C is activated by endotoxin, it hydrolyses artificial tripeptide substrate and measurable products are released, so it can be used as an alternative reagent for endotoxin analysis. Factor C gene of Tachypleus tridentatus was obtained through RT-PCR and the recombinant protein was expressed by Bac-to-Bac/BmNPV baculovirus expression system in silkworm larvae. The activity of Factor C was detected with diluted serum of silkworm larvae, and the sensitivity of endotoxin detected was 0.2 EU/mL when the serum was diluted at 1:500. The silkworm larvae expressed limulus Factor C could be used to develop a new low-cost endotoxin test reagent.
Animals ; Arthropod Proteins ; biosynthesis ; Baculoviridae ; Bombyx ; metabolism ; Enzyme Precursors ; biosynthesis ; Genetic Vectors ; Larva ; metabolism ; Recombinant Proteins ; biosynthesis ; Serine Endopeptidases ; biosynthesis