Objective To study the subchronic toxicity of sodium fluoride on mRNA expression of estrogen receptor(ER) in female mice.Methods Forty female mice were randomly divided into 4 groups according to body mass,10 in each group,and exposed to sodium fluoride solution(0,1,5,25 mg/L) through drinking water in control,low-dose,medium-dose and high-dose groups for 12 weeks.The expression levels of ERα,ER3 mRNA in ovarian tissues were determined by RT-PCR method.Results The relative expression levels of ERα,ERβ mRNA in control,low-,medium-and high-dose groups were 0.7028 + 0.0474,0.7195 ± 0.0552,0.6479 ± 0.0590,0.5684 ± 0.0513 and 0.8418 ± 0.0131,0.7729 ± 0.0974,0.7610 ± 0.0984,0.8026 ± 0.0234,respectively.The difference between high-dose and control groups of the expression level of ERα was statistically significant (P ＜ 0.05).Conclusions The subchronic toxicity of fluoride can decrease the expression of estrogen receptor in ovarian tissue,which may have a certain effect on reproductive development of female mice.

OBJECTIVETo observe the clinical efficacy of artificial skin combined with vacuum sealing drainage (VSD) in treating large-area skin defects.

METHODSTotally 18 patients with skin defects, treated with artificial skin combined with VSD from September 2008 to May 2009 in our hospital, were retrospectively analyzed in this study. There were 15 males and 3 females, aged 7-66 years, 34.3 years on average. Among them, 10 cases had skin laceration caused by traffic accidents (7 with open fractures), 1 mangled injury, 1 blast injury, 1 stump infection combined with skin defects after amputation and 5 heel ulcers.

RESULTSAll skin grafts in 16 cases survived after being controlled by VSD for one time. For the rest 2 patients, one with skin avulsion on the left foot was given median thickness skin grafts after three times of VSD, the other with open fractures in the left tibia and fibula caused by a traffic accident was given free flap transplantation. Skin grafts of both patients survived, with normal color and rich blood supply.

CONCLUSIONSkin grafting in conjunction with artificial skin and VSD is much more effective than traditional dressing treatment and worth wide application in clinic.

Adolescent ; Adult ; Aged ; Child ; Drainage ; methods ; Female ; Humans ; Lacerations ; surgery ; Male ; Middle Aged ; Postoperative Care ; Retrospective Studies ; Skin Transplantation ; Skin, Artificial ; Vacuum Curettage ; Wounds and Injuries ; surgery

OBJECTIVETo study the effects of Si-Wu-Tang on serum protein of blood deficient mice b y proteomicstechnique and study the enriching and regulating blood mechanism of Si-Wu-Tang on mocular level.

METHODThe blood deficient mice was induced by using a single dose of 3.5 Gy radiation from a 60Cogamma source, and high resolution two-dimensional polyacryamide gel electrophoresis (2-DE), computer-assisted image analysis, and mass spectrometry were used to detect regulated protein by Si-Wu-Tang.

RESULT12 lower and 4 higher protein in sera could be recovered by Si-Wu-Tang, 4 protein might be DNA-dependent protein kinase catalytic subunit, Dystrophin, KIF13A, dystonin. They play a part in DNA double-stranded break repair, recombination and modulation of transcription, transportation of mannose-6-phosphate receptor, etc.

CONCLUSIONSi-Wu-Tang can regulate serum protein in blood deficient mice, resulting in improving hematopoiesis and lessening irradiated injury.

Animals ; Autoantigens ; blood ; Carrier Proteins ; Cytoskeletal Proteins ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Dystonin ; Dystrophin ; blood ; Female ; Kinesin ; blood ; Mice ; Mice, Inbred C57BL ; Nerve Tissue Proteins ; Non-Fibrillar Collagens ; blood ; Plants, Medicinal ; chemistry ; Radiation Injuries, Experimental ; blood ; Radiation-Protective Agents ; pharmacology ; Whole-Body Irradiation

OBJECTIVE: To explore the therapeutic efficacy of manual reduction combined with percutaneous vertebroplasty in treating osteoporotic vertebral compression fractures(OVCFs) with intravertebral clefts. METHODS: The clinical data of 94 patients with osteoporotic vertebral compression fractures with intravertebral clefts treated from January 2014 to January 2017 were retrospectively analyzed. The patients were divided into group A and group B according to different operative methods. In group A, 45 patients were treated with unilateral approach PVP, including 17 males and 28 females, aged (75.35±11.82) years old, with a bone density T-value of (-4.28±0.65) g/cm³; in group B, 49 patients treated with manual reduction combined with unilateral approach PVP, including 19 males and 30 females, aged (76.79±9.64) years old, with a bone density T-value of (-4.33±0.72) g/cm³. The operation time, bone cement injection volume and postoperative complications of two groups were recorded. The VAS and ODI scores of two groups were analyzed respectively at 1, 12, 18 months after operation. Vertebral height and kyphosis Cobb angle of two groups were compared immediately after surgery and 12, 18 months after operation. The distribution of bone cement in the vertebral body was observed and its distribution excellent rate was calculated. RESULTS: There was no significant difference in operation time between two groups. The amount of bone cement injection was(8.42±1.24) ml in group A and(9.19±1.09) ml in group B, and the difference between two groups was statistically significant(<0.05). No spinal nerve root injury during operation and no complications including pulmonary embolism, bone cement toxicity and infection were found in two groups. There were 5 cases of bone cement leakage in group A and 4 cases in group B, which did not cause corresponding clinical symptoms and were not treated additionally. The distribution of bone cement in group A was excellent in 25 cases, good in 19 cases, poor in 1 case and in group B was excellent in 45 cases, good in 4 cases. The distribution excellent rate of bone cement was higher in group B than in group A (<0.05). The VAS and ODI scores before operation and 1, 12, 18 months after operation were 8.29±0.74, 2.59±0.14, 3.75±0.38, 3.84±0.88 and 40.04±3.16, 9.24±2.82, 12.27±2.64, 15.83±2.58 in group A, 8.22±0.82, 2.54±0.19, 2.81±0.23, 2.82±0.45 and 39.98±2.05, 9.16±2.10, 9.46±2.41, 9.76±2.46 in group B. There was no significant difference in VAS and ODI scores at 1 month after operation between two groups (>0.05), but group A was higher than group B at 12 and 18 months after operation (<0.05). The vertebral height and Cobb angle before surgery, immediately after surgery, and 12, 18 months after surgery in group A were(59.17±1.42)%, (85.95±2.19)%, (75.27±3.45)%, (68.34±2.24)% and(23.83±3.37)°, (15.26±2.61)°, (17.63±2.16)°, (19.46±2.54)°, and in group B were(59.31±1.87)%, (89.19±2.53)%, (88.62±2.51)%, (88.59±2.62)% and(24.72±3.78)°, (14.91±2.28)°, (15.48±2.55)°, (15.86±2.81)°. Vertebral height Immediately after surgery was greater in group B than in group A and Cobb angle in group B was smaller than in group A (<0.05). During follow-up, there was no significant change in vertebral height in group B, while vertebral body recollapse in group A(<0.05). CONCLUSIONS In the treatment of osteoporotic vertebral compression fractures with intravertebral clefts, the manual reduction combined with PVP is more effective than single PVP, which can effectively prevent vertebral body recollapse and improve the long-term efficacy of patients.
Aged ; Aged, 80 and over ; Bone Cements ; Female ; Fractures, Compression ; Humans ; Male ; Middle Aged ; Osteoporotic Fractures ; Retrospective Studies ; Spinal Fractures ; Treatment Outcome ; Vertebroplasty

OBJECTIVETo investigate the feasibility of chondrogenesis in vitro with bone marrow stromal cells (BMSCs) induced by the co-cultured chondrocytes.

METHODSThe BMSCs and chondrocytes were separated from pig and cultured. The supernatant of chondrocytes was used as the inducing solution for BMSCs from the 2nd generation. 7 days later, samples were taken and underwent immunohistochemistry and RT-PCR for detection of the expression of specific type II cartilage collagen, type II collagen and aggrecan mRNA. The cultured BMSCs and chondrocytes were mixed at a ratio of 8:2 (BMSC: cartilage cell) and were inoculated into a polyglycolic acid/polylactic acid (PGA/PLA) scaffold at the final concentration of 5.0 x 10(7)/ml. The cartilage cells and BMSCs were also inoculated separately at the same concentration as the positive and negative control. Pure cartilage cells at 20% of the above mentioned concentration (1.0 x 10(7)/ml) were used as the low concentration cartilage cell control group. Samples were collected 8 weeks later. General observations, wet weight, glycosaminoglycans (GAGs) determination and histological and immunohistochemistry examinations were performed.

RESULTSThe expression of type II collagen, type II collagen and aggrecan mRNA were positive in induced BMSCs. In the co-cultured group and the positive control group, pure mature cartilage was formed after 8 weeks of culture in vitro, and the size and shape of the scaffold were maintained. The newly formed cartilage in the two groups were almost the same in appearance and histological properties. The immunohistochemistry results indicated that the cartilage cells of the two groups all expressed ample cartilage-specific type II collagen. The average wet weight and GAG content in the co-cultured group reached more than 70% of those in positive control group. Only an extremely small amount of immature cartilage tissues was formed in local regions in pure BMSC group, and the scaffold was obviously shrunk and deformed. Although the wet weight of newly generated cartilage tissue in the low concentration cartilage cell group reached 30% of that in positive control group, the scaffold was obviously shrunken and deformed. Only regional and discontinuous cartilage tissues were formed, and the amount of newly formed cartilage was obviously less than that in the co-culture group and the positive control group.

CONCLUSIONSChondrocytes can provide a micro-environment for the formation of cartilage, and also effectively induce BMSC to differentiate into chondrocytes and form tissue-engineered cartilage in vitro.

Aggrecans ; metabolism ; Animals ; Cell Differentiation ; Cells, Cultured ; Chondrocytes ; cytology ; Coculture Techniques ; Collagen Type II ; metabolism ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Swine ; Tissue Scaffolds

Animals ; Escherichia coli ; genetics ; metabolism ; Female ; Hepatitis B Core Antigens ; biosynthesis ; genetics ; immunology ; Hepatitis B Surface Antigens ; biosynthesis ; genetics ; immunology ; Hepatitis B Vaccines ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Protein Precursors ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Vaccines, Synthetic ; immunology

OBJECTIVETo investigate the protective effects of the natural medicinal monomer isopsoralen (ISR) with estrogenic activity against oxidative damage in human lens epithelial cells B3 (HLE-B3) caused by hydrogen peroxide (H(2)O(2)) and to pursue the possible mitochondrial proteomic regularity of the protective effects.

METHODSHLE-B3 cells were treated with H(2)O(2) (300 μ mol/L), β-estradiol (E(2): 10(-8) mol/L) and H(2)O(2), ISR (10(-5) mol/L) and H(2)O(2), or left untreated. Altered expressions of all mitochondrial proteins were analyzed by protein array and surfaceenhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS). The mass/charge (m/z) ratios of each peak were tested by the Kruskal-Wallis rank sum test, and the protein peak value of the m/z ratio for each treatment by pair comparison was analyzed with the Nemenyi test.

RESULTSH(2)O(2) up-regulated the expressions of two protein spots (with m/z of 6532 and 6809). E(2) mitigated the oxidative damage, and the expression of one protein spot (m/z 6532) was down-regulated. In contrast, ISR down-regulated both of protein spots (m/z 6532 and 6809).

CONCLUSIONSISR could effectively inhibit H(2)O(2)-induced oxidative damage in HLE-B3 cells. The protein spot at m/z of 6532 might be the target spot of ISR against oxidative damage induced by H(2)O(2).

Cell Line ; Epithelial Cells ; drug effects ; metabolism ; pathology ; Estradiol ; pharmacology ; Furocoumarins ; pharmacology ; Humans ; Hydrogen Peroxide ; toxicity ; Lens, Crystalline ; pathology ; Mitochondria ; metabolism ; Oxidation-Reduction ; drug effects ; Oxidative Stress ; drug effects ; Protective Agents ; pharmacology ; Proteome ; metabolism ; Proteomics ; methods

To establish method suitable to assay HCMVpp65 of the blood donors in blood bank and to supply safe blood to the patients, the immunocytochemical techniques were used, (6 - 8) x 10(6)/ml cells were counted, 50 x 10(3) cells were detected by light microscope, The results showed that 10 positive samples in 103 samples were found, positive rate was 9.71%, among 10 positive samples, 2 samples were still positive in the second detecting. In conclusion, this method is simple, quick and effective, suitable to detect HCMVpp65 of the blood donors in the blood bank.
Blood Banks ; Blood Donors ; Female ; Humans ; Immunohistochemistry ; Male ; Phosphoproteins ; blood ; Viral Matrix Proteins ; blood

OBJECTIVETo compare different expression profiles of all known chemokine receptors in human hepatocellular carcinoma (HCC) cell lines with different metastasis potentials.

METHODSEighteen pairs of chemokine receptor primers were designed using Premier software. Expression profiles of the 18 chemokine receptors on four HCC cell lines of lower to higher potentials of metastasis (SMMC-7721, MHCC97-L, MHCC97-H and HCCLM6) were analyzed by RT-PCR. Expression of CXCR4 was detected by RT-PCR.

RESULTSExpression profiles of chemokine receptors on four HCC cell lines with different metastatic potentials had significant differences (P < 0.01), in which CCR10, CXCR4 and CXCR6 expressions decreased gradually as the metastatic potential of the cell lines increased. The expressions of CCR3, CCR4, CCR10, CCR12 and XCR1 on HCCLM6 were significantly reduced compared with SMMC-7721 (P < 0.01), whereas the expressions of CXCR1 (P = 0.006) and CXCR5 (P = 0.003) exceeded that of SMMC-7721. Except for CXCR2, CXCR6 and XCR1, most of chemokine receptors on MHCC97-H were expressed differently compared with MHCC97-L (P < 0.05), in which expressions of CCR1 (P = 0.002), CCR2 (P = 0.004) and CCR5 (P = 0.046) exceeded MHCC97-L. CXCR4 was detected only on the positive controls and SMMC-7721 when the template of total RNA was reduced one-half in RT-PCR.

CONCLUSIONChemokine receptors are expressed very differently at mRNA level on HCC cell lines with different metastatic potentials. The different profiles of chemokine receptors in tumor microenvironment and the function of CXCR4 in HCC should be further studied. Our findings have important implications in understanding the relationship between chemokine receptors and the metastatic potential of HCC.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Humans ; Liver Neoplasms ; metabolism ; pathology ; RNA, Messenger ; genetics ; Receptors, Chemokine ; metabolism

The essence of endogenous turbidity in Chinese medicine (CM) is different from cream, fat, phlegm, retention, damp, toxicity, and stasis. Along with the development of modern scientific technologies and biology, researches on the essence of endogenous turbidity should keep pace with the time. Its material bases should be defined and new connotation endowed at the microscopic level. The essence of turbidity lies in abnormal functions of zang-fu organs. Sugar, fat, protein, and other nutrient substances cannot be properly decomposed, but into semi-finished products or intermediate metabolites. They are inactive and cannot participate in normal material syntheses and decomposition. They cannot be transformed to energy metabolism, but also cannot be synthesized as executive functioning of active proteins. If they cannot be degraded by autophagy-lysosome or ubiquitin-prosome into glucose, fatty acids, amino acids, and other basic nutrients to be used again, they will accumulate inside the human body and become endogenous turbidity. Therefore, endogenous turbidity is different from final metabolites such as urea, carbon dioxide, etc., which can transform vital qi. How to improve the function of zang-fu organs, enhance its degradation by autophagy-lysosome or ubiquitin-prosome is of great significance in normal operating of zang-fu organs and preventing the emergence and progress of related diseases.
Autophagy ; Humans ; Medicine, Chinese Traditional ; Proteasome Endopeptidase Complex