The crosstalk between macrophages and bone marrow mesen chymal stem cells （BMSCs） plays a vital role in regulating tendon-bone healing. Based on the traditional Chinese medicine （TCM） theory that the kidney governs the bones and generates marrow， yin-nourishing and kidney-tonifying Chinese medicines can promote the osteogenic differentiation of BMSCs and the M1-to-M2 polarization of macrophages， thereby facilitating tendon-bone healing. A variety of related medicinal monomers （such as diosgenin and oleanolic acid）， extracts and effective fractions （such as artificial tiger bone powder and total flavonoids of Drynaria fortunei ）， as well as compound prescriptions （such as Liuwei dihuang pill， Xujin jiegu decoction and Duhuo jisheng decoction）， can modulate the crosstalk between macrophages and BMSCs by regulating the Wnt/β-catenin， nuclear factor kappa-B， phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin and transforming growth factor-β/Smad signaling pathways. They further exert anti-inflammatory， antioxidant and osteogenic effects to effectively accelerate tendon-bone healing. However， current evidence was limited to animal experiments exhibiting such as limitations insufficent alignment between animal models and clinical injury types， research focusing mostly on single signaling pathways， and unclear core active ingredients. Future studies shall optimize animal models and carry out large-sample randomized controlled trials， clarify the interactive networks of multiple signaling pathways， and isolate the core active substances of yin-nourishing and kidney-tonifying Chinese medicines， so as to advance the precise application of traditional Chinese medicines.

OBJECTIVE To explore the mecha nism by which Liuwei dihuang pill promotes tendon-bone healing by regulating the interaction between bone marrow mesenchymal stem cells （BMSCs） and macrophages through Wnt/ β -catenin pathway. METHODS BMSCs were co-cultured with macrophages at a ratio of 1∶4 and divided into control group， 10% drug-containing serum group， 20% drug-containing serum group， Wnt/ β -catenin pathway inhibitor Dickkopf-related protein 1 （DKK-1） group， and 20% drug-containing serum+DKK-1 group. After 48 hours of cell co-culture and 14 days of osteogenic induction， the CD86 and CD206 positive cell rates in macrophages， the proportion of calcium nodule positive staining area in BMSCs， the levels of osteocalcin （OCN） and oncostatin M （OSM） and alkaline phosphatase （ALP） activity in cell supernatant， as well as the mRNA and protein expression levels of OCN， Runt-related transcription factor 2 （RUNX2）， OSM， low-density lipoprotein receptor-related protein 5 （LRP5）， β -catenin in cells were detected. Pearson correlation analysis was performed to assess the correlation between LRP5 and β -catenin protein expression with OCN， RUNX2， OSM protein expression， and CD206 positive cell rate， respectively. RESULTS Compared with the control group， the CD86 positive cell rates in the 10% drug-containing serum group and the 20% drug-containing serum group were significantly reduced， while the CD206 positive cell rate， proportion of calcium nodule positive staining area， OCN level， OSM level， ALP activity， and mRNA and protein expression levels of OCN， RUNX2， OSM， LRP5， β -catenin were significantly increased （ P ＜0.05）. Compared with the 20% drug-containing serum group， the changes in the above indicators in the 20% drug-containing serum+DKK-1 group were significantly reversed （ P ＜0.05）. The protein expression of LRP5 and β -catenin was positively correlated with the pro tein expression of OCN， RUNX2， OSM， and CD206 positive cell rate. CONCLUSIONS Liuwei dihuang pill can activate the Wnt/ β -catenin pathway， induce M2 polarization of macrophages and secretion of OSM， promote osteogenic differentiation and matrix mineralization of BMSCs， and synergistically promote tendon-bone healing.

OBJECTIVE To explore the mecha nism by which Liuwei dihuang pill promotes tendon-bone healing by regulating the interaction between bone marrow mesenchymal stem cells （BMSCs） and macrophages through Wnt/ β -catenin pathway. METHODS BMSCs were co-cultured with macrophages at a ratio of 1∶4 and divided into control group， 10% drug-containing serum group， 20% drug-containing serum group， Wnt/ β -catenin pathway inhibitor Dickkopf-related protein 1 （DKK-1） group， and 20% drug-containing serum+DKK-1 group. After 48 hours of cell co-culture and 14 days of osteogenic induction， the CD86 and CD206 positive cell rates in macrophages， the proportion of calcium nodule positive staining area in BMSCs， the levels of osteocalcin （OCN） and oncostatin M （OSM） and alkaline phosphatase （ALP） activity in cell supernatant， as well as the mRNA and protein expression levels of OCN， Runt-related transcription factor 2 （RUNX2）， OSM， low-density lipoprotein receptor-related protein 5 （LRP5）， β -catenin in cells were detected. Pearson correlation analysis was performed to assess the correlation between LRP5 and β -catenin protein expression with OCN， RUNX2， OSM protein expression， and CD206 positive cell rate， respectively. RESULTS Compared with the control group， the CD86 positive cell rates in the 10% drug-containing serum group and the 20% drug-containing serum group were significantly reduced， while the CD206 positive cell rate， proportion of calcium nodule positive staining area， OCN level， OSM level， ALP activity， and mRNA and protein expression levels of OCN， RUNX2， OSM， LRP5， β -catenin were significantly increased （ P ＜0.05）. Compared with the 20% drug-containing serum group， the changes in the above indicators in the 20% drug-containing serum+DKK-1 group were significantly reversed （ P ＜0.05）. The protein expression of LRP5 and β -catenin was positively correlated with the pro tein expression of OCN， RUNX2， OSM， and CD206 positive cell rate. CONCLUSIONS Liuwei dihuang pill can activate the Wnt/ β -catenin pathway， induce M2 polarization of macrophages and secretion of OSM， promote osteogenic differentiation and matrix mineralization of BMSCs， and synergistically promote tendon-bone healing.

This paper aims to detect the antigens in porcine circovirus type 2 (PCV2) vaccines by high-performance size-exclusion chromatography (HPSEC) coupled with multi-angle laser light scattering (MALLS). With purified inactivated PCV2 and PCV2 virus-like particles (VLP) as references, two inactivated vaccines (a and b) and two VLP vaccines (c and d) for PCV2 from four manufacturers were analyzed by HPSEC-MALLS after demulsification. The antigen peaks in HPSEC-MALLS were identified by PCV2 antigen test strips, Western blotting and transmission electron microscope (TEM). The repeatability and linearity of the method were investigated. The results showed the virus antigens in the two inactivated vaccines were eluted at about 13.3 min in HPSEC. The molecular weight of these antigens was 2.61×10⁶ (±4.34%) Da and 2.40×10⁶ (±2.51%) Da, respectively, as calculated by MALLS. The antigen peaks of the two VLP vaccines also appeared at 13.3 min and the molecular weight was 2.09×10⁶ (±2.94%) Da and 2.88×10⁶ (±11.85%) Da, respectively, which was close to the theoretical molecular weight of PCV2. Moreover, an antigen peak of VLP vaccine c was observed at 11.4 min and the molecular weight was 4.37×10⁶ (±0.42%) Da. The antigen was verified to be the dimer of VLP by TEM. Vaccine d and purified Cap VLP antigens were tested repeatedly, and the RSD of the peak area (n=3) was all < 1.5%, indicating that the method was repeatable. The purified VLP were diluted in serial and tested for linearity. The result suggested good linear relationship between the peak area of VLP or VLP aggregates and the protein concentration of the sample with R² of 0.999 and 0.997, respectively. Thus, the method met the requirement for quantification and aggregate analysis. This method is accurate and efficient in in vitro quality evaluation and improvement of PCV2 vaccine.
Animals ; Antibodies, Viral ; Capsid Proteins ; Chromatography, Gel ; Circoviridae Infections/prevention & control* ; Circovirus ; Lasers ; Swine ; Vaccines, Inactivated ; Vaccines, Virus-Like Particle ; Viral Vaccines

OBJECTIVETo investigate the predictive value of serum CA125 in peritoneal metastasis, and to examine the association of CA125 with the prognosis.

METHODSClinical data of 1285 gastric cancer patients admitted to the Qilu Hospital from March 2003 to September 2008 were retrospectively reviewed. The sensitivity, specificity and accuracy of serum CA125 for peritoneal metastasis were analyzed. Clinicopathological characteristics and survival were compared between patients with normal serum CA125 level(≤35 μg/L) and patients with elevated serum CA125 level(>35 μg/L).

RESULTSThe specificity, sensitivity and accuracy of serum CA125 for peritoneal metastasis were 96.0%, 13.8% and 81.2%, respectively. A significantly higher incidence of peritoneal metastasis was observed in the patients with elevated serum CA125 level as compared to those with normal serum CA125 level(43.2% vs. 16.5%). CA125 was an independent predictor of peritoneal metastasis(RR=3.475, 95% CI:2.124-5.685). The 5-year overall survival rate in patients with elevated serum CA125 level was 13.5%, which was significantly lower than that of patients with normal serum CA125 level (49.8%, P<0.01). CA125 was an independent prognostic factor of gastric cancers(HR=2.049, 95%CI:1.355-2.873).

CONCLUSIONSerum CA125 is useful in predicting peritoneal metastasis and prognosis of gastric cancer, which should be used as a routine examination of gastric cancer patients.

CA-125 Antigen ; blood ; Humans ; Neoplasm Staging ; Peritoneal Neoplasms ; blood ; secondary ; Prognosis ; Retrospective Studies ; Stomach Neoplasms ; blood ; pathology ; Survival Rate

Objective To compare the esophageal acid and alkaline exposure characteristics of patients with cardia carcinoma after proximal gastrectomy or total gastrectomy.Method A total of 77 patients of cardia carcinoma who underwent radical resection from Sep 2007 to Sep 2011 in our hospital were retrospectively reviewed.24 hour esophageal pH monitoring were performed in all patients.Result Patients were divided into three groups:group TG had total gastrectomy (n =25),group PP had proximal gastrectomy with pyloroplasty (n =33),group NP had proximal gastrectomy (n =19).It revealed that indicators of acid reflux including the overall time length of acid episodes,＞ 5 min times of acid episodes,duration of longest acid episodes,time length of pH ＜ 4.00 and the DeMeester Scores in group NP were significantly higher than in group PP(U =32,P ＜ 0.01 ; U =35,P ＜ 0.01 ; U =23,P ＜ 0.01 ; U =39,P ＜0.01 ;U =49,P ＜0.01 respectively).Only alkline reflux was observed in group TG.The total times of alkaline episodes in PP group was significantly lower than in group TG(U =52,P ＜0.01) and group NP (U =182,P ＜0.01).＞5 min times of alkaline episodes in group TG was larger than in group PP,and that in group PP was larger than in group NP(P ＜0.01).Duration of longest alkaline episodes and total period of pH ＞7.00 in group PP was significantly higher than in group TG(U =125,P ＜ 0.01 ; U =143.5,P ＜ 0.01),and that in group TG was higher than in group NP(U =23.5,P ＜ 0.01 ; U =14,P ＜ 0.01).Conclusions Alkaline reflux deserves more attention in evaluating esophageal reflux in patients with cardia carcinoma after resection.Pyloroplasty is not helpful to relieving esophageal acid episodes while causing severe alkaline reflux.

Objective To investigate the predictive value of serum CA125 in peritoneal metastasis, and to examine the association of CA125 with the prognosis. Methods Clinical data of 1285 gastric cancer patients admitted to the Qilu Hospital from March 2003 to September 2008 were retrospectively reviewed. The sensitivity, specificity and accuracy of serum CA125 for peritoneal metastasis were analyzed. Clinicopathological characteristics and survival were compared between patients with normal serum CA125 level(≤35 μg/L) and patients with elevated serum CA125 level (> 35 μg/L). Results The specificity, sensitivity and accuracy of serum CA125 for peritoneal metastasis were 96.0%, 13.8% and 81.2%, respectively. A significantly higher incidence of peritoneal metastasis was observed in the patients with elevated serum CA125 level as compared to those with normal serum CA125 level (43.2% vs. 16.5%). CA125 was an independent predictor of peritoneal metastasis (RR=3.475, 95% CI:2.124-5.685). The 5-year overall survival rate in patients with elevated serum CA125 level was 13.5%, which was significantly lower than that of patients with normal serum CA125 level (49.8%, P<0.01). CA125 was an independent prognostic factor of gastric cancers (HR=2.049, 95%CI:1.355-2.873). Conclusion Serum CA125 is useful in predicting peritoneal metastasis and prognosis of gastric cancer , which should be used as a routine examination of gastric cancer patients.

Objective To investigate the predictive value of serum CA125 in peritoneal metastasis, and to examine the association of CA125 with the prognosis. Methods Clinical data of 1285 gastric cancer patients admitted to the Qilu Hospital from March 2003 to September 2008 were retrospectively reviewed. The sensitivity, specificity and accuracy of serum CA125 for peritoneal metastasis were analyzed. Clinicopathological characteristics and survival were compared between patients with normal serum CA125 level(≤35 μg/L) and patients with elevated serum CA125 level (> 35 μg/L). Results The specificity, sensitivity and accuracy of serum CA125 for peritoneal metastasis were 96.0%, 13.8% and 81.2%, respectively. A significantly higher incidence of peritoneal metastasis was observed in the patients with elevated serum CA125 level as compared to those with normal serum CA125 level (43.2% vs. 16.5%). CA125 was an independent predictor of peritoneal metastasis (RR=3.475, 95% CI:2.124-5.685). The 5-year overall survival rate in patients with elevated serum CA125 level was 13.5%, which was significantly lower than that of patients with normal serum CA125 level (49.8%, P<0.01). CA125 was an independent prognostic factor of gastric cancers (HR=2.049, 95%CI:1.355-2.873). Conclusion Serum CA125 is useful in predicting peritoneal metastasis and prognosis of gastric cancer , which should be used as a routine examination of gastric cancer patients.

Objective To investigate the clinicopathological characteristics and prognosis of serumb alpha-fetoprotein(AFP)-producing gastric cancer.Methods A total of 1396 cases of gastric cancer in our hospital from 2002 to 2007 were retrospectively reviewed.The relationship between serum AFP with clinicopathological factors,survival rate,prognosis and liver metastasis were analyzed.Results There were 90 AFP-producing gastric cancer patients accounting for 6.45％ of all the gastric cancer cases during the time period.In those patients there was bigger tumor size,more T3 and T4 invasion (82.2％ vs.69.2％),higher incidence of lymph node metastasis (80.0％ vs.60.2％)and liver metastasis(6.67％ vs.1.68％),and later clinical stage than that in the AFP-negative group.The 1,3,5 year-overall survival (OS) rate in AFP-positive group were 52.2％,31.1％,20％ while those in AFP-negative group were 82.5％,59.9％,48.8％.The 1,3,5 year-disease free survival(DFS) rate in AFP-positive group were 47.8％,24.4％,15.6％ while those in AFP-negative group were 77.5％,54.8％,44.3％.The AFP-positive group had a shorter median time period than the AFP-negative group when liver metastasis happened (5 months vs.11 months).Conclusions Serum AFP-producing gastric cancer was higher malignancy,it was deeper in invasion and later in clinical stage,more patients with lymph nodes and liver metastasis,low survival rate and poorer prognosis.

Objective To study retrospectively the relation of the number of all dissected and negative lymph nodes (LNs) to the prognosis of patients with stage Ⅲ rectal carcinoma after radical resection.Methods From 2002 to 2007,412 sage Ⅲ rectal carcinoma patients undergoing radical resection were enrolled.Patients were divided into five groups according to the number of dissected LNs as follows:1 to 6 lymph nodes,7 to 12 lymph nodes,13 to 18 lymph nodes,19 to 24 lymph nodes,and more than 24 lymph nodes.The association with the survival was analyzed.The Kaplan-Meier method was used to estimate survival as a function of time,and survival differences were analyzed with the log-rank test.The correlation between all dissected and negative lymph nodes was analyzed.The Cox proportional hazard model were used to investigate the risk factors for stage Ⅲ rectal carcinoma.Results The 1,3 and 5 years survival rates were respectively 79.9％,59.2％ and 43.0％.The 5-year survival rates increased with the increasing number of the examined LNs and the negative LNs,the differences were significant (20.0％、26.5％ 、43.9％ 、54.2％ 、53.5％,P =0.001 ; 10.3％ 、34.8％ 、51.9％ 、56.8％ 、70.8％,P =0.000).There were 7301 LNs dissected among which 5698 were pathology negative.The dissected LNs were correlated positively with negative LNs on the Pearson's correlation test(correlation coefficients r =0.899).The total number of dissected LNs and negative LNs were independent prognostic predictors.Conclusions The total number of dissected lymph node and negative lymph nodes are significantly correlated to prognosis of staged Ⅲ rectal carcinoma patient.On premise of standard procedure,we see all dissected and negative lymph nodes as a prognostic auxiliary index.