Efficacy, safety and influencing factors of intra-calf muscular injection of bone marrow mononuclear cells in the treatment of type 2 diabetes mellitus-induced lower extremity vascular disease.

The efficacy, safety and impact of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) associated with the intra-calf muscular injection of bone marrow mononuclear cells (BMMCs) in the treatment of type 2 diabetes mellitus (T2DM)-induced lower extremity vascular disease (LEVD) were evaluated. Patients with T2DM-LEVD were randomly divided into a control group and BMMCs group to assess the efficacy and safety of the treatment; serum VEGF and bFGF levels were detected. The BMMCs group was divided into superior genicular artery (SGA) and inferior genicular artery (IGA) subgroups as well as low-dose and high-dose subgroups for the comparison of efficacy indices. The BMMCs group exhibited significantly improved indices (P<0.05) compared with the control group and no fatalities or cancer occurred. There were no significant changes in serum VEGF and bFGF levels (P>0.05). The claudication distance in the IGA subgroup was significantly greater that in the SGA subgroup (P<0.05); the low-dose subgroup and the high-dose subgroup did not demonstrate any significant differences in each index (P>0.05). BMMC treatment for T2DM-LEVD was found to be safe and effective and had no significant impact on serum VEGF and bFGF levels in the short term; However, the degree of LEVD may affect its efficacy.

Efficient differentiation of neural stem cells induced by the rat bone marrow stromal cells.

Neural stem cells (NSCs) are valuable self-renewing cells that can maintain the capacity to differentiate into specific brain cell types. NSCs may repair and even replace the brain tissue, and ultimatley promoting the central nervous system regeneration. Therefore, it is important, for scientists and pjysicians, to study the method for efficient culture and differentiation of NSCs. Our previous study demonstrated that Bone Marrow Stromal Cells (BMSCs) can directly regulate the differentiation of NSCs into neurons, and soluble molecules excreted by BMSCs played a key role in this process. Hereby, we further identified the BMSCs-induced neurons could form the synapses, convey dopamine and express voltage-depend and receptor-depend calcium channels. Moreover, the extracellular signal-regulated protein kinase ERK1/2 pathway was founded to be involved in the process of neuron differentiation and proliferation by the in vitro experiments. Finally, by using protein array, we, for the first time, found that the cytokine-induced neutrophil chemoattractant-3 (CINC-3, a small molecule cytokine) can promote the leukocytes invasion into the inflammation site, and have the ability to induce mesencephal NSCs into neurons. Consequently, these positive findings suggested that our BMSCs-induced culture system could provide a useful tool to investigate the molecular mechanisms of neural differentiation of NSCs, which may be benifical for neurodegenerative diseases in the near future.

Overexpression of FXYD-3 is involved in the tumorigenesis and development of esophageal squamous cell carcinoma.

OBJECTIVE: To investigate the association of FXYD-3 expression with clinicopathological variables and PINCH in patients with ESCC. PATIENTS AND METHODS: Expression of FXYD-3 protein was immunohistochemically examined in normal esophageal mucous (n = 20) and ESCC (n = 64). RESULTS: Expression of FXYD-3 in the cytoplasm markedly increased from normal esophageal epithelial cells to primary ESCC (P = 0.001). The expression of FXYD-3 was correlated with TNM stages and depth of tumor invasion. Furthermore, the cases with lymph node metastasis tended to show a higher frequency of positive expression than those without metastasis (P = 0.086), and FXYD-3 expression tended to be positively related to the expression of PINCH (P = 0.063). Moreover, the cases positive for both proteins had the highest frequency of lymph node metastasis (P = 0.001). However, FXYD-3 expression was not correlated with patient's gender (P = 0.847), age (P = 0.876), tumor location (P = 0.279), size (P = 0.771), grade of differentiation (P = 0.279), and survival (P = 0.113). CONCLUSION: Overexpression of FXYD-3 in the cytoplasm may play an important role in the tumorigenesis and development in the human ESCC, particularly in combination with PINCH expression.

Cytoplasmic expression of p33(ING1b) is correlated with tumorigenesis and progression of human esophageal squamous cell carcinoma.

p33(ING1b), a newly discovered candidate tumor suppressor gene and a nuclear protein, belongs to the inhibitor of growth gene family. Previous studies have shown that p33(ING1b) is involved in the restriction of cell growth and proliferation, apoptosis, tumor anchorage-independent growth, cellular senescence, maintenance of genomic stability and modulation of cell cycle checkpoints. Loss of nuclear p33(ING1b) has been observed in melanoma, seminoma, papillary thyroid carcinoma, oral squamous cell carcinoma, breast ductal cancer and acute lymphoblastic leukemia. Inactivation and/or decreased expression of p33(ING1b) have been reported in various types of cancer, including head and neck squamous cell, breast, lung, stomach, blood and brain malignancies. Since little is known about the clinicopathological significance of p33(ING1b) in esophageal squamous cell carcinoma (ESCC), this study aimed to investigate the association of p33(ING1b) expression with clinicopathological variables and particularly interesting new cysteine-histidine rich protein (PINCH) in patients with ESCC. p33(ING1b) expression was examined by immunohistochemistry in 20 normal esophageal mucosa and in 64 ESCC specimens. The results revealed that the positive expression of p33(ING1b) protein in normal squamous cells was localized in the nucleus alone and the positive rate was 95%, while in ESCCs, the positive expression was mainly in the cytoplasm, together with nuclear expression, and the positive rate was 36% (P<0.0001). Furthermore, the cases with lymph node metastasis showed a higher frequency of positive cytoplasmic expression than those without metastasis (P=0.001). The cytoplasmic expression of p33(ING1b) was positively related to PINCH expression (P<0.0001) in ESCC, and the cases positive for both proteins had a high lymph node metastasis rate (P=0.001). In conclusion, p33(ING1b) cellular compartmental shift from the nucleus to the cytoplasm may cause loss of normal cellular function and play a central role in the tumorigenesis and metastasis of ESCC.

[Clinical effect of the concentrated suture fixation method on skin transplantation in the jaw and neck region].

OBJECTIVE: To observe the clinical effect of the concentrated suture fixation method on skin transplantation on deep burn wound or wound of cicatricial deformity after burn in the jaw and neck region. METHODS: One hundred and fourteen patients, hospitalized from April 2002 to December 2011, with deep burn or cicatricial deformity after burn in the jaw and neck region, were divided into packaging group and concentrated suture group according to the random number table. Each group had 57 patients including 48 cases with deep burn and 9 cases with cicatricial deformity. Traditional suture-package fixation method and concentrated suture fixation method were respectively used in packaging group and concentrated suture group to fix the autologous medium split-thickness skin in transplantation on wounds or scars. On post operation day (POD) 14, the skin microcirculatory perfusion flow of skin graft was measured, and the occurrence of ecchymoma, infection, and necrosis of skin in operative region were observed. The elasticity and contracture of grafted skin and scar hyperplasia on wound edge were observed 6 months after operation. Measurement data were processed with u test, while enumeration data with Fisher's exact test or Chi-square test. RESULTS: (1) On POD 14, the skin microcirculatory perfusion flow in concentrated suture group [(2.86 +/- 0.8) V] was significantly higher than that in packaging group [(2.33 +/- 0.15) V, u = 17.776, P < 0.05]. (2) Ecchymoma occurred in 4 patients of packaging group and 3 patients of concentrated suture group, but the difference between two groups was not statistically significant (chi 2 = 0.152, P > 0.05). (3) Infection in operative region was observed in 1 patient of packaging group, while no patient in concentrated suture group showed this symptom. The difference between two groups was not statistically significant (P > 0.05). (4) Grafted skin in 6 patients of packaging group showed foliated necrosis, which was not observed on those of patients in concentrated suture group. The difference between two groups was statistically significant (P < 0.05). (5) Centipede leg-like scar hyperplasia on wound edge occurred in 21 patients in packaging group and 6 patients in concentrated suture group, and the difference between two groups was statistically significant (chi 2 = 10.920, P < 0.05). (6) Poor elasticity of grafted skin was detected in 17 patients of packaging group and 4 patients of concentrated suture group, and the difference between two groups was statistically significant (chi 2 = 9.865, P < 0.05). (7) Obvious contracture of grafted skin was observed in 15 patients of packaging group and 4 patients of concentrated suture group, and the difference between two groups was statistically significant (chi 2 = 11.684, P < 0.05). CONCLUSIONS: Concentrated suture fixation method is suitable for application in transplantation of big sheet skin on wound in the jaw and neck region. It has high survival rate and is convenient for postoperative observation.

PINCH expression and its clinicopathological significance in gastric adenocarcinoma.

OBJECTIVE: Particularly interesting new cysteine-histidine rich protein (PINCH) is an important component of the local adhesion complexes and upregulated in several types of malignancies, and involved in the incidence and development of tumours. PINCH expression is also independently correlated with poorer survival in patients with colorectal cancer. However, there is no study of PINCH in gastric cancer, therefore, the aim of this project was to investigate PINCH expression and its clinicopathological significance in gastric adenocarcinoma. PATIENTS AND METHODS: PINCH expression was immunohistochemically examined in normal gastric mucous (n=30) and gastric adenocarcinoma (n=73), from gastric cancer patients. RESULTS: PINCH expression in the associated-stroma of gastric cancers was heterogeneous, and its positive rate (75%) was higher than that of normal gastric mucosa (43%, X^{2} =9.711, p=0.002). The stronger staining was observed at the invasive edge of tumour when compared to the inner area of tumour. The rate of positive PINCH (88%) in the cases with lymph node metastasis was higher than that (52%) in the cases without metastasis (X^{2}=11.151, p=0.001). PINCH expression was not correlated with patients' gender, age, tumour size, differentiation and invasion depth (p> 0.05). COMCLUSION: PINCH protein might play an important role in the tumourigenesis and metastasis of gastric adenocarcinoma.

Overexpression of MAC30 in the cytoplasm of oral squamous cell carcinoma predicts nodal metastasis and poor differentiation.

BACKGROUND: Expression of the meningioma-associated protein (MAC30) was increased in several types of tumors, including esophageal, gastric and colon tumors, compared to normal tissue. MAC30 expression levels gradually increased from normal colorectal mucosa to primary colorectal cancer and colorectal cancer spreading to the lymph nodes. MAC30 expression was related to survival in patients with colorectal cancer. However, there is no study on MAC30 in oral squamous cell carcinoma (OSCC). METHODS: Therefore, MAC30 expression in OSCC was investigated and possible associations of MAC30 expression with clinicopathological variables in OSCC have been analyzed. MAC30 expression was immunohistochemically examined in 20 normal oral mucosa and 43 OSCC specimens. RESULTS: Expression levels of MAC30 in the cytoplasm markedly increased from normal oral epithelial cells to primary OSCC. Strong cytoplasmic staining was significantly higher in primary OSCC compared to normal oral mucosa samples (51 vs. 20%, p = 0.019). Furthermore, MAC30 expression levels in primary tumors of patients with lymph node metastasis exceeded levels in those without metastasis (65 vs. 35%, p = 0.048), and MAC30 expression in poorly differentiated tumors was higher than in well-differentiated ones (90 vs. 39%, p = 0.005). CONCLUSION: Overexpression of MAC30 in the cytoplasm of OSCC may predict nodal metastasis and poor differentiation.

PINCH protein expression in normal endometrium, atypical endometrial hyperplasia and endometrioid endometrial carcinoma.

BACKGROUND: Particularly interesting new cysteine-histidine rich protein (PINCH), as an adapter protein of the LIM family for signal transduction in the integrin and growth factor pathway, is upregulated in the stroma of several common types of cancers and involved in promoting tumor progression. In the present study, we examined PINCH expression in normal endometrium, atypical endometrial hyperplasia and endometrioid carcinoma, and further studied the relationships of PINCH expression with clinicopathological variables in cancer patients. METHODS: PINCH expression was examined by immunohistochemistry in 23 normal endometrial samples, 18 atypical endometrial hyperplasias and 48 endometrioid endometrial carcinomas. RESULTS: The PINCH expression in the stroma of cancer (71%) was significantly increased compared to either normal endometrium (17%, p < 0.0001) or atypical hyperplasia (39%, p = 0.017), along with 9 cancers that had stronger PINCH expressions at the invasive margin of the cancers compared to the inner cancers. PINCH expression in cancer was higher in the patients with hypertension (p = 0.041) and estrogen exposure time >30 years (p = 0.021). On the other hand, PINCH expression was not related to menopausal status, gravid status, blood sugar/lipid, family background of cancer, histological grade, myometrial invasion, cervical involvement, lymph nodal metastases, growth pattern, estrogen and progestogen receptors (p > 0.05). conclusion: The results suggest that PINCH seems to play a role, presently unknown, in the tumorigenesis and development of endometrial cancer that merits further study.

[Alteration in bulbar conjunctiva microcirculation and interventional effect of Pentoxifylline after high-voltage electrical burn in rabbits].

OBJECTIVE: To study the changes in bulbar conjunctiva microcirculation (BCM) and the therapeutic effect of Pentoxifylline on BCM disturbance after high-voltage electrical burn (HEB) in rabbits. METHODS: Forty-five rabbits were divided into control group (C), electrical burn group (EB), and Pentoxifylline treatment group (PT) according to random number table, with 15 rabbits in each group. Model of HEB was reproduced in rabbits from EB and PT groups with voltage regulator and experimental transformer. Rabbits in C group were sham injured with the same devices without electrification. Changes in BCM were observed with microcirculation microscope at 15 minutes before HEB and 5 minutes, 1, 2, 4, 8 hour(s) post HEB (PHM or PHH), including: (1) morphology of microvessels, such as the discernible, diameters of arterioles, venules, and capillaries, the unevenness in caliber, and ischemic area; (2) dynamic changes in microvascular blood flow, such as blood flow speed in arterioles, venules, and capillaries, erythrocyte aggregation, and microthrombi formation; (3) condition of tissues surrounding microvessel, such as bleeding and exudation. Measurement data were processed with t test; enumeration data were processed with Fisher's exact test. RESULTS: (1) Morphology of microvessel: discernible of microvessels in EB and PT groups was decreased, but that of PT group was better than that of EB group. At PHM 5, diameter of arterioles, venules and capillaries was respectively (7.3+/-2.5), (12.3+/-2.4), (3.5+/-0.7) microm in EB group, all narrower than those of the control group [(14.6+/-3.1), (27.2+/-3.5), (9.0+/-1.4) microm, with t value respectively 5.23, 13.66, 14.04, P values all below 0.05]. Diameters of the microvessels in PT group [(10.2+/-3.8), (21.5+/-3.1), (7.1+/-1.2) microm] were larger than those in EB group (with t value respectively 2.21, 8.99, 10.18, P values all below 0.05). Diameters of arterioles, venules and capillaries in EB and PT groups recovered to the before HEB size at PHH 1. From PHH 2 to 8, arterioles and capillaries decreased gradually in caliber, venules dilated gradually in EB and PT groups, but the changes in PT group were not obvious. Thickness of microvessel was observed uneven in EB group at PHM 5, which lasted until PHH 8. Ischemia of the tissue was observed in EB group at PHM 5, which improved at PHH 2. Situation in PT group was better. (2) Dynamic changes in microvascular blood flow: at PHM 5, blood flow speed in arterioles, venules and capillaries was respectively (202+/-53), (198+/-44), (46+/-12) microm/s in EB group, all slower than those of the control group [(544+/-37), (359+/-32), (220+/-19) microm/s, with t value respectively 20.47, 11.51, 30.02, P values all below 0.05], and those of PT group [(335+/-42), (260+/-35), (119+/-23) microm/s] were faster than those of EB group (with t value respectively 7.55, 4.26, 14.85, P values all below 0.05). Blood flow speed in EB and PT groups recovered to the before HEB level at PHH 1. From PHH 2 to 8, blood flow speed decreased gradually in EB and PT groups, but that of PT group was faster than that of EB group. Erythrocyte aggregation in venules and capillaries was observed in EB group at PHM 5, which eased up at PHH 1, but aggregated at PHH 2, lasting until PHH 8. Obvious microthrombi were observed in EB group at PHH 2, which increased gradually. These changes were less obvious in PT group. (3) Condition of surrounding tissues of microvessel: in EB group, exudation was observed around microvessels at PHH 1, bleeding at PHH 2, with a worsening tendency. Changes in those in PT group were less obvious. CONCLUSIONS: HEB causes disturbance in BCM, but it can be ameliorated by Pentoxifylline.

[Effects of traditional Chinese medicine on the differentiation of mesenchymal stem cells to osteoblasts].

OBJECTIVE: To investigate the effects of traditional Chinese medicine on the differentiation of mesenchymal stem cells (MSC) to osteoblasts. METHODS: Six male Beagle dogs weighed 10-15 kg each were divided into three groups, group A: medicine serum group, group B: non-medicine serum group and group C: bovine serum group. The serum of group A was obtained from the femoral artery of 2 Beagle dogs drinking equivalent dose of traditional Chinese medicine according to body surface area for 7 continuous days. The serum of group B was collected from the femoral artery of 2 Beagle dogs fed with equal volume of normal saline for 7 days. The serum of group C was fetal bovine serum. The tibia marrow was harvested from another 2 Beagle dogs and MSC were isolated and purified by density gradient centrifugation. MSC were cultured in DMEM solution with fetal bovine serum. After MSC were digested by trypsin, MSC were cultured in DMEM solution with the osteogeneic inducer, which contained dexamethasone, antiscorbutic and beta-glycerophosphate. Morphological and histological changes of the MSC were observed under an inverted microscope. Alizarin monosulfonate and nitric acid argentum staining was performed to observe the calcium deposition. MSC were curtured in DMED solution with medicine serum (group A), non-medicine serum (group B) and bovine serum (group C) respectively. The growth curve was detected by the methyl thiazolyl tetrazolium (MTT). The alkaline phosphatase (ALP) activities were detected to observe the differentiation of MSC. RESULTS: The original MSC were observed as fibroblast-like cell shapes. After the osteogeneic inducer was added, MSC were polygon cells with a few polyprocess. Calcium deposition appeared during 10-14 days and alizarin monosulfonate and Von Kossa staining presented positive. MTT results showed that the number of adherent cells of group A was more than that of group B and that of group C significantly after 6 days (P < 0.05). ALP detection proved that ALP activity of group A was more than that of group B and that of group C significantly after 5 days (P < 0.05). CONCLUSIONS: The traditional Chinese medicine promotes the differentiation of MSC to osteoblasts and osteogenesis.

Expression and significance of FXYD-3 protein in gastric adenocarcinoma.

OBJECTIVE: FXYD-3, also known as Mat-8, is a member of the FXYD protein family. It was reported that this protein can associate with and modify the transport properties of Na, K-ATPase, and may play an important role in a variety of physiological and pathological states. This protein is up-regulated in certain types of cancers (such as breast, prostate and pancreatic cancer), but down-regulated in other types of cancers (such as colon and kidney cancer). No study has been performed in gastric cancer; therefore, the aim of this project was to investigate FXYD-3 expression and its clinicopathological significance in gastric adenocarcinoma. PATIENTS AND METHODS: FXYD-3 protein was examined by immunohistochemistry in normal gastric mucous (n= 29) and gastric adenocarcinoma (n=51), obtained from surgical resection of gastric cancer patients. RESULTS: FXYD-3 protein was present in the cytoplasm of normal gastric epithelial cells or gastric cancer cells. The rate of FXYD-3 strong expression was significantly higher in cancer (51% of 51) than in normal mucosa (10% of 29, X;{2}=13.210, p < 0.0001). FXYD-3 expressed strongly in ulcerative/infiltrating types of cancers compared to polypoid/fungating ones (X;{2}=5.765, p=0.016). However, FXYD-3 expression was not correlated with patient's gender, age, tumor size, lymph node status and histological grade (p > 0.05). Conclosion: Up-regulated expression of FXYD-3 protein may be involved in tumourgenesis and invasion of gastric adenocarcinoma.