58 embryos at 1, 2, 3 and 4 cell and morula stage were recovered from the oviduct and the uterine horn of the Ha-bai, Du-ha and Du-chang-ha pigs, 24 hours, 2 or 3 days of pregnancy, and were observed with SEM and TEM. The zona pellucida of 1 cell embryo was removed by treatment with 2.5％ pronase for visualization of the cell surface structure.The findings are as follows:1. The surface of the 1 cell stage embryo was densely populated with microvilli and sperm can be found on the surface with its head and tail surrounded by the microvilli and its plasma membrane of the equatorial segment of acrosome intact.2. Many spermatozoa were observed both on the surface and inside the zona pellucida of the 2 celled embryo.3. By the 4 cell stage, some of the spermatozoa had penetrated through the zona and got into the perivitelline space. 4. There were still some spermatozoa on the zona surface of the morula.5. The TEM observation revealed that the heads of supernumerary sperm were located within the cytoplasm of the blastomeres of the embryos from 1 to 4 cell stages and still not decondensed. Some of these sperm heads were actually found within the phagocytic vacuoles of the 4 celled embryos. The ultrastructure of these polyspermic embryos was normal.These results showed that polyspermy in the pig did not intefere with the cleavage of the embryo and the embryo possessed the ability to dispose the excessive sperms.

0.05).The effective rates of the two treatment schemes were 90%,87%.The cost effectiveness ratio were 108.61 and 76.35.Conclusion Homemade octreotide is better in the treatment of severe acute pancreatitis.

On 10 days of pregnancy the blastocysts of 3 mm and 9 mm in diameters wereflushed out from the uterine horns of the Du-Chang-Ha and Dong-Bei-Min pigs.The blastocysts were divided into four parts,that is the embryonic disc,the animalpole,the lateral part and the vegetal pole.The outer and inner surface of the fourparts were observed with SEM and the findings are as follows:1,The 3 mm blastocystsThe outer surface of the embryonic disc was still covered with trophoblasticcells.Three kinds of cells can be discerned:the cell with round prominences andlong microvilli(MV);the cell with thin and long MV,the cell with smooth sur-face.The trophoblast cells in other areas of the blastocyst were similar but slightlyflat.The inner surface of the blactocyst consists of endoderm cells which weredifferent in the four parts.From the embryonic disc to the vegetal pole,the surfacestructure of the endoderm cells changed regularly from small to large from dome-shaped to flat and the distance between nuclei increased gradually.The MVs on thecell surface:became less.These findings suggested that endoderm cells weremoving from the center of the embryonic disc to the vegetal pole.There weremany holes on the marginal area of the cells.The edge of the cell on the innersurface of the vegetal pole forms a network.Many round granules can be seen inthe holes of the network.Their function may be for the transport of nutrients.2,The 9 mm blastocystsThe outer surface of the embryonic disc was not covered with trophobla-stic cells but covered with ectoderm cells.The ectoderm cells in the primitivestreak had long MV on their surface but short ones on other areas of the disc.The inner surface of the embryonic disc was endoderm cells.The MVs on theendoderm cells was much more numerous than the MVs on the ectoderm cells.Thedifferentiation of the cells in the pig blastocyst was also discussed.

Objective To study the effects of electromyography-triggered neuromuscular stimulation on upper extremity functional recovery of acute stroke patients. Methods 60 stroke patients who lost wrist extension even in the electromyography (EMG) within 4 weeks were randomly assigned to experimental group and control group. The experimental group received electromyography-triggered neuromuscular stimulation, and the control group received neuromuscular stimulation. They were assessed with EMG, Fugl-Meyer assessment (FMA), Barthel index (BI) before and 4 weeks after treatment. Results The EMG, scores of FMA and BI improved in all the patients after treatment (P=0.000), and improved more in the experimental group (P<0.05). Conclusion Electromyography-triggered neuromuscular stimulation can improve the motor function of hemiplegia, even in the patients without change of EMG.

Objective To investigate the inhibitory effect of ginkgolide B (BN52021) on severe acute pancreatitis (SAP) via detecting the antagonistic effect of BN52021 on platelet-activating factor (PAF). Methods One hundred and eighty Wistar rats were randomly divided into control group (n = 60), SAP group (n = 60) and BN52021 group (n =60) according to the random number table. The 3 groups were divided into 6 subgroups at different time points after operation (1 h, 2 h, 3 h, 6 h, 12 h, and 24 h). The changes of serum amylase in each group were monitored. The expression of platelet-activating factor receptor (PAFR) mRNA and protein was detected by RT-PCR and Western blot, and the pathological changes of pancreatic tissues were observed. All the data were analyzed by one-way ANOVA. Results Serum amylase level and pathological results showed that it was successful in preparing SAP model. The serum amylase levels at postoperative hour 3, 6 and 24 were (4185 ±148) U/L, (3785 ± 124) U/L and (1360 ± 161) U/L in BN52021 group, which were significantly lower than those in SAP group [(4799 ± 107) U/L, (4920 ± 140) U/L, (2283 ± 127) U/L)]. The pathological scores at postoperative hour 3, 6, 12 were 5.95±0. 19, 5.55±0.36, 6.72±0. 30 in BN52021 group, which were significantly lower than those in SAP group (8.85 ± 0.39, 9.15 ± 0.55, 10.10 ±0. 65). The mRNA and protein expression of PAFR were gradually increased at the early stage (0.49 ± 0.09-0.71 ± 0.14 vs 0. 43 ～ O. 06-1.69 ± 0.06), and reached peak at postoperative hour 3. The expression levels of PAFR mRNA and protein in BN52021 group and SAP group at postoperative hour 3 had statistical difference among the 3 groups (F = 4.58, 6.24, P < 0.05). Conclusions The expression of PAFR mRNA and protein in the pancreatic tissue of SAP rats is dynamically changing. PAFR plays an important role in the occurrence and progression of SAP. BN52021 can reduce the expression of serum amylase and improve the pancreatic pathological changes, but it has no effect on the expression of PAFR in pancreatic tissue.

Objective To explore the pathogenic bacteria and immunologic mechanism of recurrent respiratory tract infection(RRI) in children.Methods The observation group included 50 children with RRI,26 cases were boys and 24 cases were girls, in Department of Respiratory Medicine of Wuhan Children′s Hospital were enrolled from Apr.2007 to Apr.2008.These children were divided into 3 subset groups:28 cases in 6 months-2 years old group,15 cases in 3-5 years old group,and 7 cases in 6-12 years old group.The healthy control group included 50 healthy children aged 6 months -12 years.The specimens were gathered on the next morning after the children entered hospital.The secretions of noses and pharynxes were gathered with clean and aseptic tampon from children with upper respiratory tract infection and placed in aseptic vessel,and were immediately detected with the pathogenic bacteria.The secretions of lower respiratory tract were gathered with suction method from children with lower respiratory tract infection and placed in aseptic vessel, and were immediately detected with the pathogenic bacteria,the number of superinfection with some kind of pathogenic bacteria was calculated.The children in observation group and healthy control group were exsanguinated of vein when the children were hungry to detect the cellular immunity(CD3+,CD4+,CD8+,CD4+/CD8+) and humoral immunity(IgA,IgM,IgG,C3,C4).Results Three hundred and two specimens in 6 months-2 years old group were detected,and 135 pathogenic bacteria were separated,the ratio of positive was 42.2%,and the number of superinfection was 25.One hundred and thirty-seven specimens in 3-5 years old group were detected,and 47 pathogenic bacteria were separated,the ratio of positive was 34.3%,and the number of superinfection was 7.Fifty-four specimens in 6-12 years old group were detected,and 16 pathogenic bacteria were separated,the ratio of positive was 29.6%,and the number of superinfection was 2.The top 5 kinds of bacteria that those children with RRI were easily infected were Ps.aeruginosa,K.pneumoniae,S.aurens,E.coli and streptococcus pneumoniae.The CD3+,CD8+ and CD4+/CD8+ of cellular immunity in the observation group were obviously lower than those in the healthy control group(Pa

Objective Some scholars considered that dry eye is associated to serum sex level in elder female population.But,how the estrogen and/or androgen play role in dry eye is still in controversy.The goal of the present study was to investigate the effect of estrogen and androgen on lacrimal secretion and expression of apoptosis genes in the lachrymal gland in ovariectomized rat.MethodsSixty-four female Wistar rats were divided into normal contol group,sham operation group and experiment group randomly.Ovariectomy (OVX) was performed in the rats of the experiment group and only partial fat tissue in abdominal cavity was cut in the sham operation group.Lacrimal secretion (Schirmer Ⅰ test),tear film breakup time (BUT) and corneal fluorescence staining examinations were measured in all rats before and 1,2,3,4 and 5 months after the operation.Corn oil,estrogen and androgen were systemically and topically applied 5 months after the operation for six weeks in the OVX experiment group.The experimental rats were sacrificed and the lachrymal glands were obtained for pathohistological examination.The serum estrogen and androgen levels were detected before and 5 months after the operation and before death.The expressions of bax and bcl-2 were detected by immunohistochemistry in the different groups.ResultsThe serum estrogen and androgen levels were significantly decreased after OVX in comaprison with before OVX (P＜0.05).The BUT was obviously shorter in the 1 month after OVX group (P＜0.05).The result of the Schirmer Ⅰ test decreased to 50% in 3 months after OVX (P＜0.01).Corneal fluorescence staining showed positive staining 4 months after OVX and stronger staining 5 months after OVX.In the sixth week after use of androgen,the results of BUT and Schirmer Ⅰ test were considerably decreased but stronger corneal fluorescence straining was seen.However,a complete contrary outcome was found in systemic androgen treatment rats.The expression of bax in lacrimal epithelium cells was increased after estrogen treatment and declinded after androgen treatment.The expression of bcl-2 in lacrimal epithelium cells was declinded after estrogen treatment and increased after androgen treatment.ConclusionIt is supposed that decrease of lacrimal secretion in OVX rats is associated with the decrease of serum androgen.The treatment with androgen can improve lacrimal secretion and decrease the expression of apoptosis gene in lachrymal gland in ovariectomized rat.The apoptosis of lachrymal gland epithelium is one of mechanisms of dry eye.

Objective To study whether solasodine hydrochloride (SBHL) could enhance the effect of arsenic trioxide in inducing apoptosis and affecting telomerase activity in cervical cancer HeLa cells. Methods Using cell culture methods, cervical cancer HeLa cells were cultured in vitro. The optimal concentration of SBHL was determined by MTT method from 0, 10, 20, 40, 80, 160, to 320 μmol/L. HeLa cells were grown in improved RPMI1640 supplemented respectively with arsenic trioxide(5 μmol/L As2O3), As2O3(5 μmol/L)+ SBHL( 40 μmol/L) and none (control group). The growth morphology of HeLa cells was observed under phase contrast microscopy after culture for 24, 48, and 72 h. Apoptosis of HeLa cells was determined under transmission electronic microscopy. The method of MTT was used to study the cell survival percentage. The technique of flow cytometry was used to measure cell cycle and cell apoptosis percentage. The method of tartrate-resistant acid phosphatase-enzyme linked immunosorbent assay (TRAP-ELISA) was used to determine telomerase activity of HeLa cells. Results Under phase contrast microscopy, in control group HeLa cells were round, densely packed; in As2O3 group the numbers of the cells were less, cell spacing increased; in As2O3 + SBHL group the cells shrinked significantly, nuclear fragmented as a petal-like, gap became larger. Under transmission electronic microscopy, there were rich microvillus on the cell surface in control group, cell intervals clear, immature connections, and the intervals did not close. The structure of the mitochondria in the cytoplasm was integrated. Most of the chromatin in the nucleus were, euchromatin and characteristics of apoptosis with heterochromatin increased and the chromatin condensed into masses, on the boundary of nuclear membrane. The microvillud on the cell surface were ruptured and decreased in As2O3 + SBHL group. The chromatin condensed into masses. The formation of apoptotic bodies was observed. The difference was statistically significant between groups in cell survival percentage at 24, 48, 72h(x2 = 10.39 , 13.88 , 17.21,respectively, all P < 0.05). Cell survival percentage in SBHL + As2O3 group (52.80%) was significantly less than that of As2O3 group(77.51%, x2 = 9.29, P < 0.05) at 72 h. In cell cycles, the difference was statistically significant between groups in C1 phase and S phase(F = 7.46,22.14, all P < 0.05), respectively. Compared with , control group[ (41.57 ± 1.56)%, (50.45 ± 2.37)%], cell percentages in S phase in As2O3 + SBHL group[(20.06 ± 4.98)%] and As2O3 group[(27.10 ± 5.32)%] were decreased(P< 0.05 or < 0.01), while cell percentage in C1 phase was increased[(58.70 ± 5.18)%, (69.67 ± 4.17)%, P< 0.05 or < 0.01]. The difference was statistically significant between groups in apoptotic percentage of HeLa cells (F = 4.01, P < 0.05). Compared with control group[ (1.18 ± 1.40)%], apoptosis percentage was significantly increased in As2O3 + SBHL group and As2O3 group [(21.08± 1.22)%, (6.04±2.53)%, P< 0.05 or < 0.01], respectively, and As2O3 + SBHL group was higher than As2O3 group(P < 0.01). The difference was statistically significant between groups in telomerase activity (F = 21.28, P< 0.05). Telomerase activity was inhibited in As2O3 group(1.214 ± 0.621) and As2O3A + SBHL group(0.865 ± 0.284) compared to control group (2.107 ± 0.057, all P < 0.05), and telomerase activity in As2O3 + SBHL group was lower than that of As2O3 group (P < 0.05). Conclusions SBHL enhances the effect of As2O3 in inducing apoptosis in HeLa cells, which is related to its inhibiting telomerase activity in HeLa cells.

Background The pathogenesis of dry eye is complicated,hormone level is thought to be one of impact factors in the development of dry eye.The regulation of the synthesis process of metalloproteinases(MMPs) in tissue has been reported.However,the effects of hormone on expression of MMP-2 in lachrymal gland is not clear.Objective This study was to investigate the effects of estrogen and androgen on the expression of MMP-2 in lachrymal gland in ovariectomized rats,and explore the role of MMP-2 in dry eye.Methods Sixty-four 3-monthold clean female Wistar rats were randomized into control group(8 rats),sham operation group(8 rats)and experiment group(48 rats).Ovariectomy(OVX) was performed on the rats of experiment group,and only fat tissue of abdominal cavity was cut off in the rats of the sham operation group.After 5 months of OVX,the experimental rats were subdivided into model control group,vehicle group,estrogen and androgen systemic or topical utilization groups and 8 rats for each group.Six weeks after administration of the drugs,the lachrymal gland was obtained.The expression of MMP-2 mRNA in the lachrymal gland was detected by reverse transcription PCR(RT-PCR),β-actin mRNA was used as an internal control,and the expression of MMP-2 protein was detected by Western blot,GAPDH was used as protein loading control.The use and care of the rats complied with the ARVO Statement.Results The expression of MMP-2 mRNA was strongest in the systemic estrogen group and was weakest in the systemic androgen utilization group.A significant difference in the MMP-2 mRNA expression was found among the 8 groups(F=18.60,P＜0.01),and the MMP-2 mRNA was significantly higher in the model group than that of the normal control group(0.66±0.10vs.0.47±0.10)(q=3.01,P＜0.05).In addition,the MMP-2 mRNA was significantly higher in the systemic estrogen group compared with the model group (0.83 ±0.10 vs.0.66-0.10) (q =2.79,P＜0.05) ; while the expression of MMP-2 mRNA was significantly declined in the systemic androgen group in comparison with the model group(0.12±0.04 vs.0.66±0.10)(q=11.41,P＜0.01).The MMP-2 protein presented with a strongest expression in the systemic estrogen utilization group and a weakest expression in the systemic androgen utilization groups.The expression level of the MMP-2 protein in the lachrymal gland was significantly different among the 8 groups(F =7.28,P＜0.01).The MMP-2 in the model group was significantly higher than that of the normal group(0.55±0.13 vs.0.38±0.08) (q =2.39,P＜0.05),and that in the systemic estrogen group was increased in comparison with the model group(0.69±0.12 vs.0.55±0.13) (q =1.85,P＜0.05).However,the MMP-2 in the systemic androgen group was significantly lowed in comparison with the model group(0.27±0.07 vs.0.55±0.13) (q =4.32,P＜0.01).Conclusions Estrogen may up-regulate the expression of MMP-2 in lachrymal gland,but the effect of androgen is opposite.Hormone level may play an important role in the regulation of the function of lachrymal gland.

Objective To investigate the changes of serum interleukin 8(IL-8) level in children with thyroid disfunction and its clinical significance in children with autoimmune thyroid disease(AITD).Method Serum IL-8 of 30 children with hyperthyroidism,16 children with hypotyroidism and 20 health children were detected by double antibody enzyme linked immunossorbent assay(ELISA).Results Serum IL-8 levels in hyperthyroidism and hypothyroidism children were significantly higher than that in controls(P0.05).With the remission of the diseases, IL-8 levels decreased; there was a significant difference between serum IL-8 levels in patients before and after therapy (P