Objective The aim of this study was to observe the growth difference and expression of cytochrome C of skin hair follicles in neonatal mice .Methods The morphology of different skin hair follicles of neonatal mice ( postnatal day 1-9)were observed by HE staining histology and cytochrome C was detected by immunohistochemistry .Results The skin hair follicles in different parts of neonatal mice showed differences not only in morphology but also in developmental pe -riods.Hair follicle growth in the back and tail skin had a nonlinear and growing period .After the nonlinear and growing pe-riod they began to grow rapidly .The tail development was slightly slower than that on the back .The hair follicles of vibris-sae were very special , and started to develop without a stable period .Conclusions The results of morphological observa-tion and cytochrome C immunohistochemistry demonstrate that differences exist in the hair follicle morphology and develop -mental times in the skin of different parts of the body in neonatal mice .

Objective To research the relationship between the expression of PAI-1 and the clinical characteristics of the endometrial carcinoma.Methods We detected the level of the serum PAI-1 by ELISA in the patients with endometrial carcinoma,the patients with endometrial hyperplasia and the patients with normal endometrium.The expression of PAI-1 in endometrial carcinoma and normal endometrial tissues was observed by immunohistochemistry.Results The concentration of serum PAI-1 in patients with endometrial carcinoma was 18.64 ± 6.22 μtg/L,significantly higher than those of patients with endometrial hyperplasia (6.94 ± 2.87) μg/L and patients with normal endometrium (6.68 ± 2.13)μg/L (P=0.00).The expression rate of PAI-1 was 68.2％ (15/22) in endometrial carcinoma tissue,and 8.3％ (2/24) in normal endometrial tissue (P=0.00).Compared with the early endometrial carcinoma,the expression rate of PAI-1 in the advanced endometrial carcinoma was significantly higher (6/6 w 9/16,P=0.03).Conclusions The expression level of PAI-1 may be related to the stage of the endometrial carcinoma.

Objective To investigate the multi-resistance to antibiotics of clinical isolated klebsiella pneumoniae.Methods The resistance to antibiotics of clinical isolated klebsiella pneumoniae were monitored.The discconfirmatory test was used to detect extended-spectrum β-lactamases(ESBLs) and cefoxitin three-dimension was used to detect AmpC β-lactamases.Results Among the isolates there were 53 strains of ESBLs-producing bacteria (49.5% ), 30 strains of AmpC-producing bacteria(28.0%), 24 strains of ESBLs + AmpC-producing bacteria (22.46%).They were high resistance to aminoglycosides,quinolones and cephalosporins.Conclusion The multi-resistance to antibiotics of clinical isolated klebsiella pneumoniae were widespread.It is important to control nosocomial infection to strengthen the detection of the epidemiology of ESBLs and AmpC β-lactamases in clinical isolates.

Objective To introduce the treatment efficacy of using allograft muscle ligament anatomical to rebuild anterior cruciate ligament (ACL) of knee joint under the arthroscopy.Methods Sixty-two cases patients with ACL rupture in anterior cruciate ligament reconstruction under arthroscopy.Allograft ligaments were used as graft,a bone tunnel was established in the proximal tibia and distal femur,and the graft was fixed by the extrusion screw.After the operation,the knee joint was fixed for 12 weeks,and the subjective evaluation was carried out according to the Lysholm and Larson knee score standards;in order to assess the stability of the ligament and the functional recovery of the knee joint,objective evaluation was carried out according to Lachman test in patients.Results The preoperative average Lysholm scale was (43.1±2.1) points,the final average score of 2 years after the reconstruction of the ligament was (91.0+2.3) points,there was significant difference (t=3.460,P=0.001).The preoperative average Larson scale was (41.0±2.9) points,the final average score of 2 years after the reconstruction of the ligament was (90.1±3.5) points,there was significant difference (t=3.232,P=0.001).Lachman test results were negative in 62 patients at the end of the review.No serious postoperative complications occurred,no knee infection,deep vein thrombosis and stiffness.All the patients can be completely straight 1 year after operation,knees up to 120 degrees.All patients were satisfied with the function at the end of the follow-up,no joint instability,no re-rupture occurred during the follow-up period.Conclusion Using the allogeneic single beam anatomy of anterior cruciate ligament reconstruction under arthroscopy can obtain satisfactory clinical efficacy.

ObjectiveTo explore the effects of glycomacropeptide (GMP) in human milk and formula milk on proliferation ofbifidobacterium infantis and their dose-response relationship.Methods Casein was isolated from the milk of 30 healthy postpartum women from Guangzhou Women and Children's Medical Center in September 2014, and hydrolyzed by rennet to obtain GMP, which was then purified by ultrafiltration and ion exchange chromatography. Human milk GMP and cow milk GMP (0, 250, 500, 1 000, 1 500, 2 000 and 3 000 mg/L) were added tobifidobacterium infantis liquid medium, and cultured under anaerobic conditions. Concentration of bacteria was measured by turbidimetric microplate assay (detection of OD600 nmvalue of medium). Difference of proliferative activities ofbiifdobacterium infantis in human milk GMP and cow milk GMP was compared with independent samplest-test.ResultsPurified human milk GMP concentration was 1 712.20 mg/L, with a purity of 80.3%. Increasing the cow milk GMP initial concentration in the culture medium at 250-2 000 mg/L could increase the concentration and proliferation rate ofbiifdobacteria infantis. When cultured at 36 h with GMP of various concentrations, the proliferation ofbiifdobacteria infantis maintained at a logarithmic phase. Therefore, 36 h was chosen as the test time point to compare the proliferation ofbifidobacterium infantis. At 36 h, when GMP in the medium was 1 000, 1 500, 2 000 and 3 000 mg/L, concentrations ofbiifdobacteria infantis in human milk GMP were 2.255±0.036, 2.583±0.088, 2.877±0.080 and 3.219±0.081, respectively, which were significantly higher than those in cow milk GMP (2.115±0.053, 2.312±0.064, 2.542±0.090 and 2.894±0.076;t=4.867, 5.569, 6.192 and 6.516; allP<0.01).Conclusions Both human milk GMP and cow milk GMP can promote the proliferation ofbiifdobacterium infantisin vitro, and the proliferative activity in human milk is greater than in cow milk at the same concentration of GMP.

OBJECTIVE:To determine the retative bioavailability of Diclofenac sodium eye-ointment in rabbit eyes and to evaluate its pharmacokinetics following topical application METHODS:48 rabbits were divided into two groups 0 1% Diclofenac sodium eye-ointment and Diclofenac sodium eyedrops were applied topically to rabbit eyes and aqueous humor and iridic tissue were collected at given times The concentration of Diclofenac sodium in tissue were measured by HPLC RESULTS:Diclofenac sodium eye-ointment group had higher concentration and longer half-time than Diclofenac sodium eyedrops group,however,the peak time were the same in two groups The relative bioavailabilities of Diclofenac sodium ointment in aqueous humor and iridic tissue were 183 33% and 205 68% respectively,compared with those of Diclofenac sodium eyedrops CONCLUSION:Diclofenac sodium eye-ointment can be absorbed well and released slowly,which can reduce the frequency of application of drug It is especially suitable for operation patient or application at night

Objective To investigate the effect of cyanidin-3-glucopyranoside extracted from Chinese bayberry on proliferation and apoptosis of human gastric cell line SGC7901.Methods After cocultured with C3G on different concentrations,cell proliferation was determined by MTT assay; morphology of apoptosis were observed by laser confocal microscopy; TUNEL assay was applied to measure the apoptoic rate; The expression of Bcl-2,Bax,Caspase-3,ICAD protein were observed by Western blot assay.Results C3G significantly inhibited the proliferation of SGC7901 cells in a concentration-and timedependent manner as measured by MTT method(P ＜ 0.01).After cells were treated with C3G,the presence of typical morphological changes of apoptosis was confirmed with laser confocal microscopy after Hoechst 33258 fluorescence staining.TUNEL assay indicated that the number of apoptotic cells in C3G-treated group was greater than that in the gastric cancer cells group(P ＜ 0.01).The expression level of Bcl-2 was down-regulated while the expression level of Bax was up-regulated by C3G,the ratio of Bcl-2 protein and Bax protein decreased.C3G may accelerate the activation of procaspase-3 and down-regulate the expression of ICAD(P ＜ 0.01).Conclusions C3G inhibits SGC7901 cell growth and induces apoptosis in a concentraion-and time-dependent manner.This action may be mediated by down-regulating Bcl-2/Bax,resulting in Caspase-3 activition and decreased ICAD protein expression.

BACKGROUND: Human transforming growth factor-β1 gene can be used for gene therapy of the degeneration of intervertebral discs, but the key to the experiment is to construct its effective vector.OBJECTIVE: To determine whether or not adult degenerated intervertebrai disc cells cultured in vitro after transfected by eukaryotic expression vector can express the product of human transforming factor-βl, and to provide the experimental basis of gene therapy for intervertebral disc degeneration.DESIGN: Single sample experiment. SETTING: Traumatic Orthopedic Institute of Shandong Province and the Orthopedic Department of Weihai Municipal Hospital.MATERIALS: The experiment was conducted at the laboratory of Traumatic Orthopedic Institute of Shandong Province between October 1999and January 2001. Intervertebral disc samples were from the operated patients with protrusion of irtervertebral disc after the patients were informed.Sample 1 was intervertebral disc at L4/5 from a 30-year-old woman; sample 2 was intervertebral disc at L5/S1from a 30-year-old woman.METHODS: ① Culture of adult degenerated intervertebral disc cells:Samples ex vivo were taken back to the laboratory within 30 minutes; fibrous ring cells and myelin nucleus cells cultured primarily were collected.② Transfection: Cells were put in the 24-well culture plate with 5.5×105cells in each well. Constructed PCI-hTGF-β1 eukaryotic expression vector was used to perform transfection, then transfected PCI group and nontransfected group were set. ③ The expression product of cells transfected for 48 hours was determined with immunohistochemical staining method.MAIN OUTCOME MEASURES: Comparison of absorbance of the positive cell product of eukaryotic expression vector PCI-hTGF-β1 in the primary fibrous ring cells and myelin nucleus cells in each group.RESULTS: ① Sample 1: The absorbance of positive cell product of eukary otic expression vector PCI-hTGF-β1 in the primary fibrous ring cells and myelin nucleus cells was 3.49 and 3.69 times that in PCI group, and 3.55times that in non-transfected group. ② Sample 2: The absorbance of positive cell product of eukaryotic expression vector PCI-hTGF-31 in the primary fibrous ring cells and myelin nucleus cells was 3.56 and 3.46 times that in PCI group, and 3.43 times and 3.33 times that in non-transfected group.CONCLUSION: PCI-hTGF-31, as the effective eukaryotic expression vector in the transfection of transforming growth factor-31 gene to culture degenerated intervertebral disc cells in vitro, can transfect adult degenerated intervertebral disc cells cultured in vitro and obtain the high expression of human transforming growth factorβ1 gene.

Objective To investigate the application value of identification of the scalp surface locations of cerebral ischemia lesions before direct revascularization for moyamoya disease and to design surgical approaches according to this by using the fusion of single photon emission computed tomography ( SPECT) cerebral perfusion imaging with CT imaging. Methods The clinical data of 13 adult patients with ischemic-type moyamoya disease underwent superficial temporal artery-middle cerebral artery bypass surgery were analyzed retrospectively. SPECT cerebral perfusion imaging was fused with CT imaging of the same machine before procedure. The lesions of ischemia were located on the cortical surface. The surgical approaches were designed at the center of the ischemic lesions. The patients were followed up for 6 to 12 months after procedure. The improvement of clinical symptoms and cerebral perfusion of the patients were observed after operation. Results One patient had perioperative cerebral hyperperfusion syndrome,and the others did not have any perioperative complications. At one-month follow-up, the improvement of symptoms in 4 patients were excellent,in 5 were good,in 4 were fair,and none was poor. At 6 to 12 month follow-up,the improvement of symptoms in 9 patients were excellent,in 4 were good,and none was poor. The postoperative visual SPECT cerebral perfusion imaging analysis suggested that the cerebral perfusion was improved significantly as compared with before procedure in all patients. Quantitative analysis:There was significant difference in target ischemic lesions between preoperative Fb and postoperative Fb ([2. 13±1. 06]% vs. [4. 13±2. 09]%;P<0. 05). There was significant difference between preoperative Fb and Fe ([2. 46±1. 97]% vs. [2. 13±1. 06]%;P<0. 05). The postoperative BFCR was [67. 57±3. 78]%( >50%) , which indicated that the efficacy of the procedure was remarkable. The superficial temporal arteries fed to brain of the patients were observed after procedure by using the head CT angiography. The postoperative head MRI reexamination showed no new infarcts occurred at 6 months. Conclusion Combine SPECT cerebral perfusion imaging with CT imaging to design surgical approach for superficial temporal artery-middle cerebral artery bypass surgery may improve the efficacy and reduce the risks of operation.

Objective To investigate the effects of lipopolysaccharides (LPS) in different concentrations on the proliferation and interleukin(IL)-6,IL-1 β and tumor necrosis factor-α(TNF-o) secretion of intestinal epithelial cells (IEC-6) of rats in vitro.Methods IEC-6 of rats were divided into normal group (0 mg/L,group A),0.1 mg/L group (group B),0.5 mg/L group (group C),1.0 mg/L group (group D),5.0 mg/L group (group E) and 10.0 mg/L group(group F).Different groups cells were exposed to LPS with different concentrations for 3 h,5 h and 7 h.Thiazolyl blue(MTT) was performed to investigate the proliferation of IEC-6.The concentrations of IL-6,IL-1 β and TNF-α in culture supernatant were detected by enzyme linked immunosorbent assay(ELISA).Results The proliferation rate of IEC-6 was gradually lower while the concentration of LPS increased.After co-culture with LPS 3h and 5 h,the proliferation rates of group B,group C,group D,group E and group F had no significant difference with those of group A (all P ＞ 0.05);after co-culture with LPS 7 h,the proliferation rates of group B,group C,and group D had no significant difference with those of group A (all P ＞ 0.05);the proliferation rates of group E and group F had significant difference with those of group A(t =4.216,P =0.014;t =14.991,P =0.000).The proliferation rates of group E and group F were lower after co-culture with LPS 5 h than 7 h,and there were significant differences (t =2.576,P =0.033;t =2.975,P =0.018);but there was no significant differences between group E and group A after co-culture with LPS 5 h (P ＞ 0.05).Group B,group C,group D,group E and group F all had a significant higher level of IL-6 than group A after co-culture with LPS 3 h,5 h and 7 h(all P ＜0.01).In addition,group E had the highest level of IL-6 at all time points.And the peak level of IL-6 rose after co-culture with LPS 5 h.The TNF-α level and IL-1 β level of group B,group C,group D,group E and group F all had no significant differences than that of group A after co-culture with LPS 3 h,5 h and 7 h (all P ＞ 0.05).Conclusions In a certain concentration,incubation time range,the proliferation rates of IEC-6 cells were gradually lower while the concentration of LPS increased.Co-cultured IEC-6 cells with LPS(0-10.0 mg/L) can stimulate them secrete to IL-6.The highest level of IL-6 was of group E after 5 h co-culture.LPS had no effects on TNF-α and IL-1 β level of IEC-6 cells cultural supernatant.So 5.0 mg/L concentration of LPS stimulating IEC-6 cells for 5 h can be chosen to build the IEC-6 inflammatory models.