Objective To explore a method for isolation and culture of human epidermal stem cells. Methods Epidermis was obtained by digesting human foreskin with Dispase Ⅱ and Trypsin-EDTA.After suspension on the epidermal stem cell medium (ESCM), these single epidermis cells were inoculated onto human collagen Ⅳ-coated flasks and cultured at 37 ℃ in a humidified atmosphere containing 5% CO_2 for 10 min. The nonadherent cells were rinsed off 10 min after inoculation, and the adherent cells continued to be cultured after enriching and abstraction by type Ⅳ collagen. The cell growth was observed through inverted microscope, and the cell cloning efficiency and time of clone sustain were also detected. Immunocytochemistry was used to observe the expression of ?_1-integrin and keratin 19(K19). Keratinocytes were served as controls. Results It was revealed by histological observation that colonies were formed 24 hours after inoculation. The isolated and cultured cell cloning efficiency was higher and the time of clone sustain was longer than that of the control group. Positive expression of ?_1-integrin and K19 of cultured cells was detected by immunocytochemistry. Conclusion Adult epidermal stem cells could be successfully isolated and cultured by adhension with type Ⅳ collagen and culture with ESCM.

OBJECTIVETo evaluate the effect of early application of high-volume hemofiltration treatment (HVHF) on the levels of lactic acid, pro-inflammatory cytokines and C-reactive protein (CRP) in plasma, as well as APACHE II score in patients suffering from severe sepsis.

METHODSThirty patients meeting the diagnosis of severe sepsis were enrolled in the trial within 24 hours of insults. The level of lactic acid, interleukin-6 (IL-6) and CRP in plasma were measured before HVHF and at 24, 48 or 72 h following HVHF treatment.

RESULTThe plasma levels of lactic acid and IL-6 decreased significantly at 24 h, 48 h, 72 h after HVHF (P <0.05), while, IL-10 did not differ significantly following HVHF (P>0.05), when compared with that before HVHF.

CONCLUSIONThe early application of HVHF could clear the plasma lactic acid and pro-inflammatory cytokines, and improve the tissue oxygenation in severe sepsis.

APACHE ; Adult ; C-Reactive Protein ; analysis ; Female ; Hemofiltration ; methods ; Humans ; Interleukin-10 ; blood ; Interleukin-6 ; blood ; Lactic Acid ; blood ; Male ; Middle Aged ; Sepsis ; blood ; therapy ; Treatment Outcome ; Young Adult

OBJECTIVETo compare the early clinical results of thoracoscopic and mini-open thoracotomic anterior correction for idiopathic thoracic scoliosis.

METHODSTwenty-three cases with idiopathic right thoracic scoliosis were divided into 2 groups. Group A includes 8 females with average age of 14.8 years and average Cobb angle of 54 degrees . The Risser sign was +++ approximately ++++. These patients were operated on with thoracoscopic Eclipse instrumentation. Group B covers 2 males and 13 females with average age of 13.8 years and average Cobb angle of 57 degrees . The Risser sign was ++ approximately ++++. These patients were operated on with mini-open thoracotomic anterior instrumentation. The operative time, blood loss, postoperative drainage, instrumented levels, curve correction and early loss of correction of both groups were analyzed.

RESULTSThe patients of group A had average operative time of (360 +/- 72) min, (629 +/- 145) ml of intra-operative blood loss, (7.4 +/- 1.1) of instrumented levels, (500 +/- 150) ml of post operative drainage, 74 +/- 14% of curve correction rate and (8.6 +/- 2.7)% of early loss of correction after 6 approximately 18 m follow-up. The patients of group B had average operative time of (246 +/- 64) min, (300 +/- 110) ml of intra-operative blood loss, (7.8 +/- 0.9) of instrumented levels, (210 +/- 90) ml of post operative drainage, (70 +/- 12)% of curve correction rate and (4.6 +/- 1.9)% of early loss of correction. The curve correction rates of thoracoscopic and mini-open thoracoscopic anterior correction were not significantly different (P >0.05). But the operative time, blood loss, postoperative drainage, and early loss of correction showed significant difference (P <0.05).

CONCLUSIONSThoracoscopic and mini-open thoracotomic anterior correction for idiopathic thoracic scoliosis have their own indications. Both techniques are safe and effective to correct the idiopathic thoracic scoliosis with satisfied early results. But the early loss of correction of mini-open thoracotomic anterior correction is significantly less than that of thoracoscopic anterior correction.

Adolescent ; Adult ; Female ; Follow-Up Studies ; Humans ; Male ; Minimally Invasive Surgical Procedures ; Scoliosis ; surgery ; Spinal Fusion ; methods ; Thoracic Vertebrae ; surgery ; Thoracoscopy ; Thoracotomy ; methods ; Treatment Outcome

OBJECTIVETo observe the effect of Suyu capsule on behavior, injury of hippocampal neurons and Ca2+ ion in hippocampal synaptic in the depression model mice.

METHODSixty male Kunming mice were randomly divided into 5 groups, the control group, the model group and three Suyu capsule groups (the doses were 22.8, 11.4, 5.7 g x kg(-1) respectively). The model was established by separation and chronic unpredictable mild stimulation. The increased weight and crossing score, rearing score were measured by open-field and sweet water consumption of mice. Cone cell and configuration of neuron in CA1, CA3 region of hippocampus were observed by Nissl. The concentration of hippocampal synaptic Ca2+ ion was detected by fluorimetry.

RESULTComparing with the mice of control, the increased weight was slowered ( P < 0.01), the scores of rearing and crossing were decreased (P < 0.01), sweet water consumption were decreased too (P < 0.01), numbers of cone cell in CA3 region of hippocampus were decreased obviously (P < 0.01), and Ca2+ ion in hippocampal synaptic was increased obviously. Comparing with the mice of model, Suyu capsule (22.8 g kg(-1)) could increase the increased weight on the 14th and 21 st day obviously (P < 0.05); Suyu capsule (22.8 g x kg(-1)) could increase the scores of crossing obviously (P < 0.05), Suyu capsule (22.8, 11.4 g x kg(-1)) could increase the scores of rearing obviously (P < 0.01, P < 0.05); Suyu capsule (22.8, 11.4, 5.8 g x kg(-1)) could increase sweet water consumption obviously (P < 0.01, P < 0.05, P < 0.05; Suyu capsule (22.8, 11.4, 5.8 g x kg(-1)) could increase numbers of cone cell in CA3 region of hippocampus obviously (P < 0.01, P < 0.05, P < 0.05); Suyu capsule (22.8, 11.4, 5.8 g x kg(-1)) decreased Ca2+ ion in hippocampal synaptic with dose-effect relationship (P < 0.01, P < 0.01, P < 0.05).

CONCLUSIONSuyu capsule can improve all the symptoms of the depression model mice and protect injury of hippocampal neurons in the depression model mice. The possible mechanism of action is to restrict Ca2+ ion overfreight.

Animals ; Antidepressive Agents ; pharmacology ; Behavior, Animal ; drug effects ; Body Weight ; drug effects ; Calcium ; metabolism ; Capsules ; Depression ; metabolism ; pathology ; physiopathology ; Dose-Response Relationship, Drug ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Hippocampus ; pathology ; Male ; Mice ; Neurons ; metabolism ; Neuroprotective Agents ; pharmacology ; Plants, Medicinal ; chemistry ; Random Allocation ; Synapses ; metabolism

OBJECTIVETo observe the therapeutic efficacy of Shenfu Injection (SFI) on patients with severe sepsis and its effects on serum levels of interleukin-6 (IL-6) and interleukin-10 (IL-10).

METHODSSixty-eight patients with severe sepsis were randomly assigned to the SFI group (36 cases, treated by SFI + routine therapy) and the control group (32 cases, treated by routine therapy). The acute physiology and chronic health evaluation II (APACHE II) score and Marshall score were observed before treatment, 3 and 7days after treatment. The therapeutic efficacy was assessed, and the 28th-day mortality rates were compared. The serum levels of IL-6 and IL-10 were determined by enzyme-labeled immunosorbent assay (ELISA) before and after treatment. C-reactive protein (CRP) was determined by immunoturbidimetric assay.

RESULTSThere was no significant difference in the APACHE II score, Marshall score, IL-6, IL-10, or CRP between the two groups before treatment (P>0.05). APACHE II score and Marshall score of all patients decreased after treatment, with more obvious decrease shown in the SFI group (P<0.05). The mortality rate in the SFI group and the control group was 25.0% (9/36) and 37.5% (12/32) respectively, with no significant difference shown between the two groups (P>0.05). The serum levels of IL-6 and CRP obviously decreased after 7 days of treatment (P<0.05). But more decrement was shown in the SFI group, showing significant difference when compared with the control group (P<0.05). There was no significant difference in the serum IL-10 level between the two groups before and after treatment (P>0.05).

CONCLUSIONSFI could lower the serum IL-6 level, regulate the equilibrium of proinflammatory factors and anti-inflammatory cytokines in severe sepsis patients, thus playing a role in improving the therapeutic efficacy.

APACHE ; Adult ; Aged ; C-Reactive Protein ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Interleukin-10 ; blood ; Interleukin-6 ; blood ; Male ; Middle Aged ; Phytotherapy ; Sepsis ; blood ; drug therapy

OBJECTIVETo detect the prevalence of spinal cord abnormality in "idiopathic" left thoracic scoliosis and analyze its clinical significance.

METHODSFrom October 1997 to October 2003, 59 "idiopathic" left thoracic scoliosis were treated, including 31 males and 28 females. The age was from 7 to 44 years with an average of 15 years. The Cobb angle was from 15 degrees to 108 degrees with average 56 degrees . The holocord MRI was made for all patients.

RESULTSThirty-three patients were found cord abnormality, including 24 Chiari malformation with syringomyelia, 5 syringomyelia, 1 Chiari malformation with syringomyelia and diastematomyelia, 1 Chiari malformation with syringomyelia and tethered cord, 1 syringomyelia with tethered cord, and 1 Dandy-Walker malformation. The prevalence of spinal cord abnormality in left thoracic scoliosis was 56%. Compared to the left thoracic scoliosis without cord abnormality, the left thoracic scoliosis with cord abnormality had been found more in males and the patients with bigger Cobb angle (P < 0.05).

CONCLUSIONSFor left thoracic scoliosis, especially male patient or patient with bigger Cobb angle, associated cord abnormality should be highly suspected. Preoperative holocord MRI is very meaningful for this kind of patients.

Adolescent ; Adult ; Child ; China ; epidemiology ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Prevalence ; Scoliosis ; complications ; surgery ; Spinal Cord Diseases ; diagnosis ; epidemiology ; etiology ; Thoracic Vertebrae

OBJECTIVETo construct a lentiviral expression vector of the PIAS-NY gene, and establish a mouse spermatocyte-derived cell line with a stable overexpression of PIAS-NY.

METHODSPIAS-NY was synthesized, amplified by PCR and cloned into the lentiviral vector expression plasmid pGC-FU. After digestion and sequencing, pGC-FU-PIAS-NY, pHelper 1.0 and pHelper 2.0 were co-transfected into 293T cells. Then the lentiviral particles were used to transfect the mouse spermatocyte-derived cells. The expression of the PIAS-NY protein was detected by Western blot.

RESULTSWe successfully constructed the lentiviral expression vector pGC-FU-PIAS-NY and established a mouse spermatocyte-derived cell line with a stable overexpression of PIAS-NY.

CONCLUSIONThe construction of the lentiviral expression vector pGC-FU-PIAS-NY and the obtainment of stably transfected mouse spermatocyte-derived cells have paved the way for further studies on the roles of the PIAS-NY gene in spermatogenesis.

Animals ; Cell Line ; Genetic Vectors ; Lentivirus ; genetics ; Male ; Mice ; Plasmids ; Protein Inhibitors of Activated STAT ; genetics ; Spermatocytes ; cytology ; Transfection

OBJECTIVETo study the change of melatonin receptor mRNA expression in paravertebral muscles in adolescent idiopathic scoliosis (AIS), and analyze its relationship with the etiology of AIS.

METHODSTwenty cases with average age of (15.1 +/- 2.2) years and average Cobb angle of 56 degrees +/- 16 degrees , including 10 cases with Cobb angle > 50 degrees and 10 cases with Cobb angle

RESULTSThe MT2 mRNA expression of paravertebral muscles on concave side was higher than that on convex side in AIS and CS groups (P < 0.05), but the MT1 mRNA expression showed no significant difference (P > 0.05). In AIS group, the ratio of MT2 mRNA expression on concave side compared with convex side in cases with Cobb angle > 50 degrees and cases with Cobb angle 0.05). The MT1 and MT2 mRNA expression showed no significant difference in control group (P > 0.05).

CONCLUSIONSThe melatonin receptor expression in bilateral paravertebral muscles in AIS is asymmetric, which may be a secondary change and has no important role in the pathogenesis of AIS.

Adolescent ; Adult ; Case-Control Studies ; Child ; Female ; Humans ; Male ; Muscle, Skeletal ; metabolism ; RNA, Messenger ; genetics ; Receptors, Melatonin ; biosynthesis ; genetics ; Scoliosis ; metabolism

OBJECTIVETo study the histochemical changes of muscle fibers and motor end-plates of paravertebral muscles, and analyze their relationship with the etiology of scoliosis associated with syringomyelia as compared with adolescent idiopathic scoliosis (AIS) and non-scoliotic patients.

METHODSAll the enrolled patients were divided into three groups: Group I consisted of 20 patients with scoliosis associated with syringomyelia, Group II included 16 patients with AIS, and Group III included 10 patients without scoliosis. Bilateral biopsy of paravertebral muscles was performed during scheduled spinal surgery. HE staining, nicotin-lateral biopsy of paravertebral muscles was performed during scheduled spinal surgery. HE staining, nicotinamide adenine dinucleotide hydrogen-tetrazolium reductase ( NADH-TR), and alpha-naphthyl acetate esterase staining techniques were used for histological evaluation. Neurogenic and myogenic pathological changes and changes of motor end-plates of paravertebral muscles were compared among these three groups.

RESULTSNeurogenic pathological changes of muscle fibers were found in 12 (60% ) patients in Group I but was not found in Group II and III. The numbers of both T0 type motor end-plates and pathological end-plates on the convex side were significantly larger than those on the concave side in Group I ( P < O. 05 ). In Group II , however, the numbers of both T0 type motor end-plates and pathological end-plates on the concave side were significantly larger than those on the convex side (P < 0.05). No significant difference was found between two sides in Group III.

CONCLUSIONThe histochemical changes of paravertebral muscles in patients with scoliosis and syringomyelia are different from those in AIS patients. It is suggested that a primary denervation of paravertebral muscles exist in scoliosis associated with syringomyelia, which may play a role in the pathogenesis of scoliosis.

Adolescent ; Adult ; Case-Control Studies ; Child ; Female ; Humans ; Male ; Motor Endplate ; pathology ; Muscle Fibers, Skeletal ; pathology ; Muscle, Skeletal ; pathology ; Scoliosis ; complications ; pathology ; Staining and Labeling ; Syringomyelia ; complications ; pathology

BACKGROUNDBenign prostate hyperplasia is one of the most common diseases affecting the health of the aging males. Watchful waiting is an acceptable management strategy for benign prostate hyperplasia in which the patient is monitored by the physician but receives no active intervention. The epidemiological data on this are lacking in China. Our study was designed to evaluate the changes of signs and symptoms of patients with benign prostate hyperplasia during management by watchful waiting in China.

METHODSOne hundred and forty-five patients with benign prostate hyperplasia aged > 50 years were enrolled in management by watchful waiting. All the patients were visited every 6 months and were given an International Prostate Symptom Score and Quality of Life questionnaire to complete. They also had uroflowmetry and were assessed using ultrasonography to get the volume of prostate, transition zone and amount of residual urine. The Student's t test, the Chi-square test, and variance analysis were used in the statistical analysis.

RESULTSAll patients were visited after 6 months, the mean volume of transitional zone was found to have increased by 1.6 ml (P < 0.01), International Prostate Symptom Score was increased by 0.8 (P < 0.01) and Quality of Life was increased by 0.2 (P < 0.01), and there was no statistical change in other data. Among these patients, 17.9% (26/145) visited again after 12 months when the data failed to show a statistically significant difference among the three groups (0, 6, and 12 months).

CONCLUSIONSAfter one year's follow-up, the progression of benign prostate hyperplasia was slow and the clinical data did not undergo much change.

Aged ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Prostatic Hyperplasia ; psychology ; therapy ; Quality of Life