BACKGROUND:Adipose-derived mesenchymal stem cells have the ability to self-renew and have pluripotent potential under specific conditions in vitro, which have broad application prospects in clinical practice. However, isolation and culture of adipose-derived mesenchymal stem cells stil appear to have many difficulties and shortcomings. OBJECTIVE:To isolate and culture rabbit adipose-derived mesenchymal stem cells in vitro in order to study their morphology, cellsurface markers and biological properties as wel as to investigate the osteogenic and adipogenic potentials of adipose-derived mesenchymal stem cells in vitro. METHODS:Primary adipose-derived mesenchymal stem cells were isolated from the subcutaneous adipose tissue of posterior cervical region from New Zealand white rabbits and digested by 0.1%col agenase I. The cells were passaged and amplified by the trypsin digestion. The passage 4 adipose-derived mesenchymal stem cells were induced to differentiate after exposure to adipogenic or osteogenic medium. The oil red O staining, alkaline phosphatase and alizarin red staining were used to detect the results. The cellviability was detected by the cellcounting kit 8 method to drawn the growth curve. cellsurface markers were examined using flow cytometry. RESULTS AND CONCLUSION:The adipose-derived mesenchymal stem cells isolated from the subcutaneous adipose tissue of rabbits exhibited a fusiform adherent growth in a vortex pattern, and had a strong capability of proliferation that could be passaged stably to the 10th generation. Flow cytometry results showed that the cells highly expressed CD29, CD90, CD44, but lowly expressed CD45 and CD34. After adipogenic induction, the adipose-derived mesenchymal stem cells were positive for oil red O staining;after osteogenic induction, the cells were both positive for alkaline phosphatase and alizarin red staining. These findings suggest that the adipose-derived mesenchymal stem cells were successful y isolated and cultured from the subcutaneous adipose tissue of rabbits, and these cells are pluripotent with the potential to differentiate into adipocytes and osteoblasts, which are expected to be ideal seed cells for bone tissue engineering.

Objective To explore the clinicopathological characteristics and prognosis of invasive micropapillary carcinoma of the breast (IMPC),and the distinction between IMPC and invasive ductal carcinoma of the breast (IDC).Methods From February 2004 to November 2013,195 IMPC patients and 420 IDC patients were analyzed retrospectively.Results There were significant differences in mammilla invasion,lymph vessel invasion,orange peel sign,soft tissue encroachment,neoadjuvant chemotherapy,radical mastcctomy,lymph node metastasis,clinical stages,tumor size,lymph node staging,estrogen receptor (ER),progestin receptor (PR),human epidermal growth factor receptor 2 (HER2),molecular subtyping,ratio of radiation,ratio of endocrine therapy,disease-free survival (DFS),overall survival (OS)between the two groups,all P ＜0.05.Patients with IMPC had lower 5-year DFS and OS rates (68.2％ and 73.8％,respectively) than IDC patients (85.7％ and 88.6％,respectively),all P ＜ 0.05.In IMPC patients with positive ER/PR,HER2-negative,smaller tumor volume,less lymph node metastasis,negative nipple invasion,negative lymphatic vessel tumor thrombus,negative orange peel change had higher 5-year DFS and OS rates than those with negative ER/PR,HER2 overexpression,larger tumor volume,more lymph node metastasis,positive nipple invasion,positive lymphatic vessel tumor thrombus,positive orange peel change,all P ＜ 0.05.Besides,the patients with pathologic stage Ⅰ had higher OS than those with stage Ⅲ (P ＜ 0.05).Cox regression analysis found that orange peel change,lymph vessel invasion and HER2 were the independent risk factors for the survival time of patients with IMPC.Conclusions IMPC patients have lower DFS and OS compared with IDC.

Objective To investigate the clinicopathologic characteristics and prognosis of medullary breast carcinoma.Methods We conducted a retrospective analysis on clinical and pathologic data of 166 patients with medullary breast cancer.Results All the patients were female with a median age of 52 years old.The proportion of patients with stage Ⅰ,Ⅱ and Ⅲ disease was 16.9％,68.1％,15.0％,respectively.The Luminal,HER-2 overexpressing and triple-negative subtypes constituted 31.3％,8.4％,and 60.3％,respectively.There was significant difference in regional lymph node status of medullary breast cancer patients with different molecular types (x2 =18.248,P =0.003),but not in tumor size,TNM stage,histological grade,and expression of Ki67 (all P ＞ 0.05).Multivariate survival analysis indicated that TNM stage was an independent predictor in the prognosis of medullary breast cancer (HR =5.664,P =0.001).All the patients were followed up from 15 months to 145 months with a median follow-up time of 108 months.The 5-year survival rate was 91.5％ and the 10-year survival rate was 87.2％.Conclusions The prognosis of medullary breast cancer is favorable.Personalized treatment according to the TNM stage and histopathologic characteristics achieve a favorable prognosis.

Objective To investigate the clinical and pathologic features of squamous cell carcinoma of the breast.Methods The clinical and pathologic data of 23 squamous cell carcinoma of the breast patients admitted between 1984 and 2013 to Tianjin Medical University Cancer Hospital was analyzed retrospectively.Results Primary squamous cell carcinoma of the breast was a very rare tumor accounting for 0.06％ of all breast cancers.All of the 23 patients were females aged 28 years to 87 years(median age 49 years).Average tumor size was 4.5 cm.9 patients suffered from lymph node metastasis at admission (39.1％).The positive rates of estrogen receptor (ER),progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER-2) were 13.6％ (3/23),4.5％ (1/23) and 0 (0/20) respectively.With a follow-up time varying from 5 months to 36 months recurrence or metastasis were found in 8 patients,and another 1 patient was found having distant metastasis at admission.Lung metastasis (7/9) was most common.6 patients died.Conclusions Squamous cell carcinoma of the breast is highly invasive,with low rate of positive receptors and early distant metastasis or recurrence after operation,and poor patients' survival.

Objective To identify the protein interacting with hepatitis E virus(HEV) recombi-nant capsomeric particles(P239). Methods Protein interacting with HEV was analyzed by the pull-down, MALDI-TOF-MS, co-immunoprecipitation (Co-IP) and CONFOCAL. Results A protein interacting with HEV recombinant particle (P239) was identified as HSP90 by MALDI-TOF-MS. The interaction between HSP90 and P239 was further confirmed by Co-IP. The protein level and localization of HSP90 and P239 in HepG2 were detected. The total quantity of HSP90 didn't change, and the movement of HSP90 from plasma membrane to perinuclei region with P239 was observed. Conclusion HSP90 may play an important role in the trafficking of P239. It suggests that HSP90 participate in the transportation of HEV after infection, which may contribute to the prevention and control of the disease.

Objective To explore the anatomic basis for and clinical outcomes of the internal fixation which preserves the pronator quadratus (PQ) for distal radius fractures.Methods Twenty cadaveric specimens of adult upper extremity were used for this study (14 males and 6 females).The radial and ulnar lengths of PQ,the distal and proximal widths of PQ,the distances from the distal end of PQ to the articular surface of the distal radius and to the transverse line of the wrist,and the width of the bony tunnel of PQ were dissected and measured to study the anatomical features of PQ.A retrospective study was conducted of the 18 distal radius fractures which had been treated from March 2015 to March 2017 by internal fixation with T-shaped anatomic locking compression plate (LCP) with PQ preserved.They were 8 males and 10 females,with an average age of 52.7 years (range,from 28 to 65 years).According to the AO classification,there were 8 cases of type 23-A,5 ones of type 23-B and 5 ones of type 23-C1.The functional outcomes of the wrist were assessed using the Cooney scoring system at the last follow-ups.Results The PQ muscle was flat and like a right angle trapezoid with rich blood vessels.The radial and ulnar lengths of PQ were about 4.60 cm and 4.46 cm;the distal and proximal widths of PQ were about 4.41 cm and 4.48 cm;the distance from the distal end of PQ to the transverse line of the wrist was about 3.61 cm;the widths of the distal and proximal bony tunnels were about 3.08 cm and 1.91 cm.The 18 patients were followed up for 6 to 36 months (average,11.5 months).Bone union was achieved in all the patients after a mean time of 2.5 months (range,from 2 to 3 months).The mean Cooney score for the wrist function was 97.7 (range,from 95 to 100) at the last follow-up,yielding an excellent rate of 100％.Conclusions The transverse line of the distal radius fracture is located between 1/4 and 1/2 of the distal PQ.The bony tunnel of PQ is wide enough.It is feasible to preserve the distal PQ muscle in the internal fixation of distal radius fractures of types 23-A,23-B and 23-C1,because it may lead to rapid recovery of the patients and satisfactory wrist function.

Deficiency, stasis, water and toxin are of great significance in the pathogenesis and pathologic evolution of chronic heart failure (CHF). Based on "deficiency, blood stasis, water and toxin", the pathogenesis and treatment of CHF were discussed in this article. It was found that in the pathogenesis, deficiency--deficiency of heart qi and deficiency of heart yang were the origin of the disease, and blood stasis, water and toxin were the markers of the disease. Among them, blood stasis was the central pathological link, and also an important mechanism that could aggravate the disease and cause a vicious cycle; water-phlegm and water dampness were the basic pathological products; toxin-heat toxin, water toxin, and stasis toxin were the final results of disease progress and product accumulation. In terms of treatment, CHF can be divided into four stages: early, middle, late and end. In the early stage, tonifying qi and regulating heart can be used for the treatment of root cause, and promoting blood circulation and water can be used for the treatment of symptoms; tonifying qi and yin and reinforcing the healthy qi, reducing blood stasis, purging turbid, and eliminating pathogenic factors can be used in the middle stage; reducing blood stasis and removing toxic materials should be used in the late stage, supplemented with warming yang and increasing urine excretion; astringing yang,generating body fluids, tonifying qi and yang should be used in the end stage. At the same time of treating by stages, attention should be paid to adhering to a holistic concept and dialectical treatment; pay attention to timing and flexible medication; adopting a combination of Chinese and Western approaches and integrating them.

Objective: To investigate the seroprevalence and epidemiological characteristics of hepatitis E virus (HEV) infection in pregnant women in Xiamen. Methods: Sera samples of 910 pregnant women were collected from September 2014 to June 2015 in Xiamen Huli District Maternity and Child Care Hospital. Those who intended to give birth in target hospital were included in a subgroup which was asked to collect the second serum sample. All samples were tested for anti-HEV IgM and IgG antibody by enzyme linked immunosorbent assay (ELISA). HEV RNA was tested by reverse transcription-polymerase chain reaction (RT-PCR) for the positive samples of anti-HEV IgM antibody, meanwhile, the quantitative detections for anti-HEV IgG were conducted in specimens positive for anti-HEV IgG. Results: Of the 910 pregnant women, 8 (0.88%, 95%CI 0.45%-1.73%) were anti-HEV IgM positive. HEV RNA was found in 3 cases through RT-PCR and viral load values were between 600 and 700 copies/ml; 140 (15.38%, 95%CI 13.19%-17.68%) were anti-HEV IgG positive and geometric mean concentration of the samples was 0.385 Wu/mL (95%CI 0.332-0.445 Wu/ml). The positive rate of anti-HEV IgG increased with age (P=0.004). In the subgroup, 150 pregnant women were included and followed up, 4 of those were defined as 'new HEV infection cases’ and the incidence was evaluated as 10.7/100 person-year (95%CI 3.39-25.7/100 person-year). Conclusions There were a low percentage of HEV carriers in pregnant women in Xiamen, but the risk of new primary infection in pregnant women during pregnancy was much higher than the general population, suggesting that it is necessary to expand sample size to clarify the burden of HEV infection during pregnancy.

Osteoarthritis (OA), in which M1 macrophage polarization in the synovium exacerbates disease progression, is a major cause of cartilage degeneration and functional disabilities. Therapeutic strategies of OA designed to interfere with the polarization of macrophages have rarely been reported. Here, we report that SHP099, as an allosteric inhibitor of src-homology 2-containing protein tyrosine phosphatase 2 (SHP2), attenuated osteoarthritis progression by inhibiting M1 macrophage polarization. We demonstrated that M1 macrophage polarization was accompanied by the overexpression of SHP2 in the synovial tissues of OA patients and OA model mice. Compared to wild-type (WT) mice, myeloid lineage conditional Shp2 knockout (cKO) mice showed decreased M1 macrophage polarization and attenuated severity of synovitis, an elevated expression of cartilage phenotype protein collagen II (COL2), and a decreased expression of cartilage degradation markers collagen X (COL10) and matrix metalloproteinase 3 (MMP3) in OA cartilage. Further mechanistic analysis showed thatSHP099 inhibited lipopolysaccharide (LPS)-induced Toll-like receptor (TLR) signaling mediated by nuclear factor kappa B (NF-κB) and PI3K-AKT signaling. Moreover, intra-articular injection of SHP099 also significantly attenuated OA progression, including joint synovitis and cartilage damage. These results indicated that allosteric inhibition of SHP2 might be a promising therapeutic strategy for the treatment of OA.