OBJECTIVETo investigate the differential potential of mesenchymal stem cells (MSCs) derived from human umbilical cord blood (hUCB) into insulin-secreting cells and its inducing condition.

METHODSUCB nucleated cells (NCs) were isolated and cultured in Mesencult media. The obtained UCB MSC were purified by adherence method and expanded. Then they were induced with epidermal growth factor (EGF), B-mercaptoethanol and high concentration of glucose. The induced cells were identified by RT-PCR. Intracellular insulin was examined by immunocytochemistry. The quantity of insulin secretion and glucose-simulated insulin release were examined by chemiluminescence immunoassay. The induced cells were also transplanted into renal subcapsular space of STZ-induced hyperglycemic mice to observe the in vivo lowering effect on hyperglycemia.

RESULTSThe induced cells morphologically became round and were gathering into a mass. The expression of some genes related to pancreatic islet was found by RT-PCR. Chemiluminescence immunoassay showed insulin positivity and the cells secreted a low concentration of insulin [(0.37 +/- 0.06) mU/L]. The induced cells responded to high glucose challenge with a stimulation index of 1.76. After those cells grafted into renal sub-capsule there was an in vivo lowering effect on blood glucose level on STZ hyperglycemic mice.

CONCLUSIONMSCs from UCB can differentiated into insulin secreting cells.

Animals ; Cell Differentiation ; drug effects ; Cells, Cultured ; Diabetes Mellitus, Experimental ; surgery ; Fetal Blood ; cytology ; Humans ; Insulin ; metabolism ; Insulin-Secreting Cells ; cytology ; metabolism ; transplantation ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Mice ; Mice, Nude

OBJECTIVETo explore the operative skills and effect of unilateral pedicle screw combined with contralateral percutaneous transfacet screws fixation in treating degenerative low lumbar disease.

METHODSFrom January 2009 to December 2011,22 patients with degenerative low lumbar disease were treated with transforaminal lumbar interbody fusion, during the operations, unilateral pedicle screw and contralateral percutaneous transfacet screw fixation were performed. There were 16 males and 6 females, aged from 32 to 71 years old with an average of (51.1 ± 10.6) years, including single segment in 20 cases and two segments in 2 cases. Clinical effects were evaluated according to visual analogue score (VAS) and Oswestry Disability Index (ODI).

RESULTSAll patients were followed up from 1 to 2.5 years with an average of 18 months. One case complicated with leakage of cerebrospinal fluid after operation and 1 case with lower limb pain of decompression-side on the 3rd day after operation. Twenty-two patients got bony fusion. There were no instability and evidence of instrument failure during follow-up. The VAS and ODI score decreased from preoperative 8.24 ± 0.72, 36.72 ± 6.84 respectively to 3.18 ± 0.66, 4.36 ± 1.12 at the final follow-up (P < 0.05).

CONCLUSIONUnilateral pedicle screw combined with contralateral percutaneous transfacet screw fixation is safe and feasible surgical technique in treating low lumbar degenerative disease. It has advantages of little trauma, rigid fixation, high fusion rate, and less complication. etc.

Adult ; Aged ; Biomechanical Phenomena ; Female ; Humans ; Intervertebral Disc Degeneration ; physiopathology ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Pedicle Screws ; Spinal Fusion ; methods

Background:TLR4 can mediate immune and inflammatory responses,TFF3,MUC2 are the intestinal mucosa protection factor and can maintain the intestinal barrier function. Aims:To investigate the effect of Anchang Yuyang decoction on colon tissue TFF3,MUC2 and TLR4 gene expressions in rats with ulcerative colitis. Methods:TNBS was used to establish ulcerative colitis model in rats. Ninety Wistar rats were randomly divided into normal control group,model group,low,moderate and high dose Anchang Yuyang decoction groups and mesalazine group,and distilled water,different concentrations of Anchang Yuyang decoction and mesalazine were given respectively. All the rats were sacrificed after 21 days. Colonic histopathological score was assessed,and RT-PCR was used to detect gene expressions of colon tissue TFF3, MUC2 and TLR4. Results:Compared with model group,histopathological score and TLR4 expression were significantly decreased in moderate,high dose Anchang Yuyang decoction groups and mesalazine group(P < 0. 05),expressions of TFF3 and MUC2 were significantly increased( P < 0. 05). Compared with moderate dose Anchang Yuyang decoction group,histopathological score in high dose Anchang Yuyang decoction group and mesalazine group was significantly decreased(P < 0. 05),and TFF3 expression was significantly increased( P < 0. 01). Compared with moderate dose Anchang Yuyang decoction group and mesalazine group,MUC2 expression in high dose Anchang Yuyang decoction group was significantly increased(P < 0. 01),and TLR4 expression was significantly decreased( P < 0. 01). Conclusions:Anchang Yuyang decoction can promote the repair of colonic mucosa in rats with ulcerative colitis,and its mechanism may be related to the increase of TFF3 and MUC2 gene expressions and down regulation of TLR4 gene expression.

OBJECTIVETo study human hepatocarcinoma stem cell markers.

METHODSTumor tissue samples were obtained from 8 patients with hepatic cellular cancer undergoing surgical tumor resection and the tumor cells were cultured with primary tissue culture in vitro. The cell subpopulations with each marker were isolated from the tumor cells by immunopanning using oval cell markers (CD34, c-kit, Thy-1, CK7, CK19, CK14). The cells of each phenotype were injected into nude mice to measure their ability of tumor formation and the tumor mass was weighed one month after implantation. The tumorigenic subpopulations of the cells were injected into the nude mice again in 1/4 or 1/10 of their original densities to further analyze their ability of tumor formation.

RESULTSAmong all the cell subpopulations, those positive for CD34, c-kit and CK7, respectively, showed marked difference from the negative cells in their ability of proliferation and tumor formation. The CD34(-), c-kit(+) and CK7(+) subpopulations of the cells exhibited strong capacity of tumor formation even in only 1/4 or 1/10 of their original density.

CONCLUSIONSThere are heterogeneous subpopulations within human hepatocarcinoma with observable difference in tumor formation ability. The strong tumor formation ability of CD34(-), c-kit(+) and CK7(+) subpopulation of the cells suggests that CD34(-), c-kit(+) and CK7(+) represent part of the surface markers of hepatocarcinoma stem cells.

Antigens, CD34 ; analysis ; Biomarkers, Tumor ; analysis ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Humans ; Keratin-7 ; analysis ; Liver Neoplasms ; metabolism ; pathology ; Neoplastic Stem Cells ; metabolism ; Proto-Oncogene Proteins c-kit ; analysis ; Tumor Cells, Cultured

In view of the effective traditional Chinese medicine (TCM) in the treatment of clinical depression, the mechanism is not clear, this study attempts to research the cause of depression in a complex situation to lay the foundation for the next step of TCM curative effect evaluation. Based on the brain wave of 120 depression patients and 40 ordinary person, the change regulation of acetylcholine, dopamine, norepinephrine, depression neurotransmitters and excited neurotransmitters in the whole and various encephalic regions' multi-neurotransmitters of depression patients-serotonin are analysed by search of encephalo-telex (SET) system, which lays the foundation for the diagnosis of depression. The result showed that: contrased with the normal person group, the mean value of the six neurotransmitters in depression patients group are: (1) in the whole encephalic region of depression patients group the dopamine fall (P < 0.05), and in the double centralregions, right temporal region and right parietal region distinct fall (P < 0.01); (2) in the right temporal region of depression patients group the serotonin rise (P < 0.05); (3) in the right central region, left parietal region of depression patients group the acetylcholine fall (P < 0.05), left rear temporal region fall obviously (P < 0.01). The correlation research between antagonizing pairs of neurotransmitters and neurotransmitters: (1) the three antagonizing pairs of neurotransmitters-serotonin and dopamine, acetylcholine and norepinephrine, depression neurotransmitters and excited neurotransmitters, in ordinary person group and depression patients group are characterizeed by middle or strong negative correlation. Serotonin and dopamine, which are characterized by weak negative correlation in the right rear temporal region of ordinary person group, are characterized by strong negative correlation in the other encephalic regions and the whole encephalic (ordinary person group except the right rear temporal region: the range of [r] is [0.82, 0.92], P < 0.01)/(depression patients group:the range of [r] is [0.88, 0.94], P < 0.01); acetylcholine and norepinephrine, in the whole and various encephalic region are characterized by middle negative correlation(ordinary person group:the range of [r] is [0.39, 0.76], P < 0.01 or P < 0.05)/(depression patients group: the range of [Ir] is [0.56, 0.64], P < 0.01); depression neurotransmitters and excited neurotransmitters are characterized by middle strong negative correlation (ordinary person group: the range of [r] is [0.57, 0.80], P < 0.01)/(depression patients group: the range of [r] is [0.68, 0.78], P < 0.01). (2) The two neurotransmitters which are not antagonizing pairs of neurotransmitters, serotonin and excited neurotransmitters, or acetylcholine and depression neurotra-nsmitters, or dopamine and depression neurotransmitters in the various encephalic regions are characterized by weak negative correlation. Serotonin and excited neurotransmitters are characterizeed by weak negative correlation (ordinary person group: in the right central region, left parietal region, double front temporal regions, right rear temporal region, the range of [r] is [0.25, 0.50], P < 0.01 or P < 0.05)/(depression patients group: in the whole encephalic regions, double parietal regions, double occipital regions, right front temporal region, left central region, left frontal region, the range of [r] is [0.18, 0.37], P < 0.01 or P < 0.05); acetylcholine and depression, neurotransmitters are characterized by weak negative correlation (ordinary person group: in the double frontal regions, left parietal region, left front temporal region, right rear temporal region, the range of [r] is [0.31, 0.46], P < 0.01 or P < 0.05)/(depression patients group: in double rear temporal regions, right front temporal region, double occipital regions, left central region, the range of [r] is [0.20, 0.32] , P < 0.01 or P < 0.05); do-pamine and depression neurotransmitters are characterized by weak middle negative correlation (ordinary person group: in left parietal region, right central region, left frontal region, left occipital region, double front temporal regions, the range of [r] is [0.33, 0.68], P < 0.01 or P < 0.05)/(depression patients group: in the whole region and other various regions except the left frontal region, right central region, the range of Irl is [0.21, 0.34], P < 0.01 or P < 0.05). Dopamine and acetylcholine or norepinephrine and serotonin are characterized by weak positive correlation in all encephalic regions. Dopamine and acetylcholine are characterized by weak positive correlation (ordinary person group: in left frontal region, right parietal region, left front temporal region and left rear temporal region, the range of [r] is [0.37, 0.46], P < 0.01)/(depression patients group: in the whole region and the orther various regions except the double central regions, the range of [r] is [0.23, 0.5], P < 0.01 or P < 0.05); norepinephrine and serotonin are characterized by weak positive correlation (ordinary person group: in double front temporal regions, double rear temporal regions, right frontal region and left parietal region, the range of [r] is [0.34, 0.48], P < 0.01 or P < 0.05)/(depression patients group: in the whole and various regions, the range of [r] is [0.18, 0.42], P < 0.01). The main differences between the depression patients group and ordinary person group are: (1) In the whole regin, left frontal region and right central region of depression patients group, the six neurotransmitters all fall normally (P < 0.05). (2) The percent of dopamine falling or including dopamine falling, or including dopamine falling and serotonin rising in depression patients group increases. The percent of dopamine falling or including dopamine falling in the whole region, right frontal region, right central region increases (P < 0.01), such as dopamine decreasing, serotonin increasing dopamine decreasing, serotonin increasing acetylcholine decreasing dopamine decreasing, dopamine decreasing norepinephrine increasing depression neurotransmitters decreasing, serotonin increasing acetylcholine decreasing dopamine decreasing neurotransmitters increasing and so on. (3) The percent of acetylcholine falling, or including acetylcholine falling, or including acetylcholine falling and neurotransmitters (beta)-receptor)rising in depression patients group increases. The percent of acetylcholine falling, or including acetylcholine falling in the right temporal region, double central regions increases (P < 0.05 or P < 0.01), such as acetylcholine decreasing, acetylcholine decreasing neurotransmitters increaseng, acetylcholine decreasing neurotransmitters increasing depression neurotransmitters decreasing, serotonin increasing acetylcholine decreasing dopamine decreasing neurotransmitters increasing and so on. It's showed in research that depression patients' brain are characterized by multi-neurotransmitters abnormal, the synchronous change of multi-neurotransmitters has some certain regularities, which are not the simple linear relation. It's conformed that the three antagonizing pairs, neurotransmitters-serotonin and dopamine, acetylcholine and norepinephrine, depression eurotransmitters and excited neurotransmitters of ordinary person group and depression patients group, are both characterized by strong antagonizing relation, that the two neurotransmitters which are not antagonizing pairs of neurotransmitters are characterized by weak positive correlation or negative correlation, prompt maybe has the indirect causal relationship. And the change of six neurotransmitters in depression patients' various encephalic regions is rather complex. It's conformed preliminarily that the right frontal region and right central region are characterized by dopamine decreasing, acetylcholine decreasing, serotonin increasing dopamine decreasing, serotonin increasing acetylcholine decreasing dopamine decreasing, dopamine decreasing norepinephrine increasing excited neurotransmitters decreasing, serotonin increasing acetylcholine decreasing dopamine decreasing neurotransmitters increasing, acetylchoine decreasing neurotransmitters increasing, acetylcholine decreasing neurotransmitters increasing excited neurotransmitters decreasing and so on. Contrasted with the ordinary person group, the depression patients group have the notable difference.
Acetylcholine ; metabolism ; Adolescent ; Adult ; Aged ; Brain ; metabolism ; Case-Control Studies ; Depression ; metabolism ; Dopamine ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Neurotransmitter Agents ; metabolism ; Norepinephrine ; metabolism ; Serotonin ; metabolism ; Young Adult

OBJECTIVETo investigate the effect of rat mesenchymal stem cells (MSCs) transfected with different oncogenes differentiate into hepatocellular carcinoma (HCC) in vitro.

METHODSMSCs, transfected with different oncogenes c-myc, K-ras, c-myc and K-ras and amplified in vitro, were infused into rats via vena portae. The recipient rats were divided into the hepatic impairment group, which were fed with tetrachloromethane and the healthy control group. At day 7, 14, 21, 28 and 42 following grafting, the complete livers were obtained and examined using fluorescence detection, conventional pathology and immunohistochemistry detection of GFP, c-kit and AFP to study the colonization and distribution of stem cells in rat liver.

RESULTSNo immunological rejection occurred after grafting of allogenic MSCs. The infused MSCs colonized in the recipient rat liver. Liver tumors were present in 6 rats grafted with MSCs that were transfected with K-ras, K-ras and c-myc, and the protein expression of AFP was detected using immunocytochemistry at day 7. Rats grafted with MSCs that were transfected with c-myc gene had no obvious tuberosity or tumor. Small oval cells were found microscopically in the periphery of vena portae, and immunohistochemistry staining of AFP was negative. Immunohistochemical staining of c-kit was positive in all livers of rats that were transfected with MSCs.

CONCLUSIONSHepatocellular carcinoma may derive from genetically mutated MSCs.

Animals ; Bone Marrow Cells ; cytology ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Differentiation ; Cells, Cultured ; Female ; Genes, myc ; genetics ; Genes, ras ; genetics ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Liver ; metabolism ; pathology ; Liver Neoplasms, Experimental ; metabolism ; pathology ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Microscopy, Fluorescence ; Rats ; Rats, Sprague-Dawley ; Transfection

OBJECTIVESTo compare short-term and long-term change of paraspinal muscle between percutaneous and open pedicle screw fixation in the treatment of thoracolumbar fractures.

METHODSThirty-three patients were divided into four groups: short-term percutaneous pedicle screw fixation group, short-term open pedicle screw fixation group, long-term percutaneous pedicle screw fixation group, and long-term open pedicle screw fixation group. Paraspinal muscle were studied by needle electromyography and CT. Cross-sectional area and color grade information of paraspinal muscle were measured using CT image.

RESULTSThe area and color grade of paraspinal muscle changed significantly after surgery. The color grade of paraspinal muscle showed significant change while the muscle area observed no significant change in the two short-term groups; There was significant change in paraspinal muscle area, however no significant change was found in muscle color grade of the two long-term groups. In electromyography study the results showed that there was significant difference in the two short-term groups, however no significant difference existed in the long-term groups. There was no significant difference of patients treated by the two surgical technique in long-term function evaluation.

CONCLUSIONBoth percutaneous and open pedicle screw fixation damage paraspinal muscle, however the muscle showed less injury treated by percutaneous pedicle fixation.

Adult ; Bone Screws ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; instrumentation ; methods ; Humans ; Lumbar Vertebrae ; injuries ; Male ; Middle Aged ; Muscles ; pathology ; physiopathology ; Spinal Fractures ; pathology ; physiopathology ; surgery ; Thoracic Vertebrae ; injuries ; Time Factors

Objective To investigate the change of the basic fibroblast growth factor(bFGF) leve in human detrusor muscle(DM)in bladder outlet obstruction(BOO)due to benign prostatic hyperplasia(BPH)and its implication.Methods Fifty-four patients with BPH were divided into two groups:the obstructive DM stability and instability groups;and 15 men with bladder tumor who underwent operation in the same period were enrolled in the control group.The bFGF mRNA level in DM was measured by reverse transcription and polymerase chain reaction(RT-PCR)and the bFGF protein level was measured by immunohistochemical staining method.Results The bFGF-mRNA expression level of bladder smooth muscle cells was significantly lower in the control group than that in the obstructive DM stability and instability groups(all P＜0.05),but there was no significant difference between the obstructive DM stability and instability groups(P＞0.05). Conclusions The expression level of bFGF mRNA in bladder DM is elevated in BOO due to BPH,but there is little or no correlation between the increased expression of bFGF mRNA and detrusor muscle instability.

Objective To establish the feasibility of performing an endoscopic anterior approach for upper cervical spine in a clinical setting.Methods Application of this method on 13 Chinese cadavers was conducted to verify the practicability of this technique.Anatomic data were obtained by measuring the anterior cervical specimens,and anatomic observation was conducted in the neighboring structure through below the su- perior thyroid artery.Results The superior thyroid vascular-nerve plexus was neighbor to the puncture can- nula.But there has a quite large distance between the hypoglossal,the glossopharyngeal,the lingual artery, the external branch of the superior laryngeal nerve and the puncture cannula.There has enough space between the posterior wall of the pharynx and the prevertebral fascia to put the MDE canuula.Conclusion This ca- daver and clinical study demonstrates that an endoscopic anterior approach to the upper cervical spine is safe and feasible,and can be a valid alternative to the conventional transoral approach.

BACKGROUNDStudies of interleukin (IL)-4 and IL-6 in the exhaled breath condensate (EBC) of asthmatic patients are limited. This study was to determine the effect of inhaled corticosteroid (ICS) treatment on IL-4 and IL-6 in the EBC of asthmatic patients.

METHODSIn a prospective, open-label study, budesonide 200 μg twice daily by dry powder inhaler was administered to 23 adult patients with uncontrolled asthma (mean age 42.7 years) for 12 weeks. Changes in asthma scores, lung function parameters (forced expiratory volume in 1 s [FEV1], peak expiratory flow [PEF], forced expiratory flow at 50% of forced vital capacity [FEF50], forced expiratory flow at 75% of forced vital capacity, maximum mid-expiratory flow rate) and the concentrations of IL-4 and IL-6 in EBC were measured.

RESULTSBoth asthma scores and lung function parameters were significantly improved by ICS treatment. The mean IL-4 concentration in the EBC was decreased gradually, from 1.92 ± 0.56 pmol/L before treatment to 1.60 ± 0.36 pmol/L after 8 weeks of treatment (P < 0.05) and 1.54 ± 0.81 pmol/L after 12 weeks of treatment (P < 0.01). However, the IL-6 concentration was not significantly decreased. The change in the IL-4 concentration was correlated with improvements in mean FEV1, PEF and FEF50 values (correlation coefficients -0.468, -0.478, and -0.426, respectively).

CONCLUSIONSThe concentration of IL-4 in the EBC of asthmatic patients decreased gradually with ICS treatment. Measurement of IL-4 in EBC could be useful to monitor airway inflammation in asthmatics.

Administration, Inhalation ; Adult ; Asthma ; drug therapy ; physiopathology ; Breath Tests ; Budesonide ; administration & dosage ; Female ; Forced Expiratory Volume ; Humans ; Interleukin-4 ; analysis ; Interleukin-6 ; analysis ; Male ; Middle Aged ; Peak Expiratory Flow Rate ; Prospective Studies