Aim Fuzhisan ( FZS ) , a Chinese herbal complex prescription that has been used for the treat-ment of AD for over 15 years, is known to enhance the cognitive ability in AD patients. In this study, to in-vestigate whether FZS reduces Aβ25-35-induced Tau protein hyperphosphorylation in neonatal rat cortical neurons by suppressing the cyclin-dependent kinase 5 ( CDK5 ) pathway. Methods Neonatal Wistar rats born within 24 h were selected to separate and purify their cortical neurons for culture in vitro. After 7-day culture of cortical neurons in vitro, 20 μmol · L-1 Aβ25-35 was used to act on them for 24 h. Medication groups were pretreated with FZS ( 20 mg · L-1 ) , CDK5 inhibitor roscovitine ( 15 μmol · L-1 ) and cal-pain preparation calpeptin (20 μmol·L-1 ) for 24 h, followed by reaction with 20 μmol·L-1 Aβ25-35 for 24 h. Tau protein phosphorylation levels at Ser396, Ser202 and Thr231 and the protein level of CDK5 acti-vator proteins p25/p35 were assayed by Western blot. Fluorescence intensity was measured with a fluores-cence microplate reader to reflect calpain activity. CDK5 kinase activity was assayed by immunoprecipita-tion. Results After 20 μmol·L-1 Aβ25-35 acting on cortical neurons for 24h, there were increments in the following: Tau protein phosphorylation levels at Ser396, Ser202 and Thr231, CDK5 kinase activity, CDK5 activator protein p25 level, and calpain activity. In the 20 mg·L-1 FZS treatment group, Aβ25-35 was suppressed markedly, resulting in increments in Tau protein phosphorylation levels at Ser396 , Ser202 and Thr231 , suppression of CDK5 kinase activity and p25 protein level, and elevation in calpain activity. Both CDK5 inhibitor roscovitine and calpain preparation cal-peptin, as positive control drugs, also played a role in suppressing Tau protein hyperphosphorylation. Con-clusion FZS can suppress Aβ25-35-induced Tau pro-tein hyperphosphorylation in cortical neurons through the calpain/CDK5 pathway.

AIM: To observe the effects of folic acid (FA) on antioxidant enzyme, nitric oxide synthase (NOS) and nitric oxide (NO) in ovariectomized (OVX) rats.METHODS: Forty three-month-old female SD rats were randomly divided into 5 groups:sham group, OVX group, diethylstilbestrol group (0.03 mg· kg-1· d-1), low-dose FA group (5 mg· kg-1· d-1) and high-dose FA group (20 mg· kg-1· d-1).Gastric gavage started 1 week after operation and lasted for 10 weeks.The rats in sham group and OVX group were given distilled water instead of FA as controls.At the end of the 10th week, the L5 vertebra and right femur were removed for determination of bone mineral density (BMD).The bone homogenates were made using the L3 and L4 vertebrae.The levels of the total antioxidant capacity ( TAC) , glutathione peroxidase ( GSH-Px) , malondialdehyde ( MDA ) , NOS and NO were detected in plasma and bone homogenates.RE-SULTS:Compared with sham group, the BMD levels in L5 vertebra and right femur and the levels of GSH-Px and NO in the plasma were all decreased.The levels of TAC, GSH-Px, NOS and NO in the bone homogenates were also decreased, while the MDA concentration was increased in OVX group (all P<0.01).Compared with OVX group, the levels of TAC, GSH-Px, NOS, NO and BMD of the L5 vertebra and right femur were all increased, while the MDA concentration was de-creased in high-dose FA group (all P<0.01).CONCLUSION:In female SD rats, ovariectomy leads to a significant re-duction of antioxidant enzyme, NOS and NO levels.Oxidative stress is possibly involved in the development of osteoporo-sis.Protection against osteoporosis by high-dose FA may be linked to improvement of antioxidant enzyme activity, the levels of NOS and NO as well as a reduction of oxidative stress in ovariectomized rats.

Objective To discuss parallel fibular osteoseptocutaneous flap apply in heel defect reconstruction Methods From February,2006 to December,2011,parallel fibular osteoseptocutaneous flap were used to repair calcaneus and skin defects in 4 cases.The causes included road accident in 2 cases and strangulation in 2 cases.The defect locations were at the back of the calcaneus (1/2 of calcaneus in 2 cases and 2/3 in 2 cases,respectively).Results Followed up 24-72 months,all cases achieved bone union in allograft and had no complications of absorption,infection and repulsion.The flaps survived completely in 4 cases ; the distal flaps necrosed partly in 1 case and healed by dressing.According to USA foot-malleolus scores of AOFAS,1 case was excellent,2 good,and 1 fair.Conclusion Parallel fibular osteoseptocutaneous flap reconstruction heel defect can recover patients visage and heel function,improve their life quality.

BACKGROUND:Hydrophobic acrylic foldable intraocular lens should be evaluated biologicaly with New Zealand rabbits as implant objects prior to clinical trial. OBJECTIVE:To observe the biological safety of hydrophobic acrylic foldable intraocular lens. METHODS: Twelve New Zealand rabbits were enroled. The right eyes were implanted with self-developed hydrophobic acrylic foldable intraocular lens (Shenyang Baiao Medical Device Co., Ltd., China) as experimental group, and the left eyes were implanted with Acrysof IQ SN60WF (Alcon, USA) as control group. RESULTS AND CONCLUSION: After implantation, there were no significant differences in the cornea, anterior chamber, implant position, posterior segment of eyebal between two groups. Tissue proliferation had no obviously difference between the two groups, and there were cornea and lens pouch inflammations. No macrophages and other inflammatory cels were visualized on the surface of intraocular lens, and fibrous tissues were found on the intraocular lens surface and in the haptics root. These findings suggest that the self-developed hydrophobic acrylic foldable intraocular lens has no difference from Acrysof IQ SN60WF widely used in clinic, and it has the biological safety.

Objective To investigate the cytotoxic activities of γδT cells against methotrexate (MTX)-resistant osteosarcoma (OS) cells.Methods The MTX-resistant U2OS cell line (U2OS/MTX) was established by in vitro exposing the parental cells to MTX at stepwise-increasing concentrations.Periph-eral blood mononuclear cells ( PBMCs) were isolated from healthy subjects and stimulated with zoledronate ( ZOL) in combination with IL-2 to induce the proliferation ofγδT cells.The cytotoxicity ofγδT cells against U2OS/MTX cells was analyzed by using a standard lactate dehydrogenase release assay (LDH).Flow cy-tometry analysis and ELISA were performed to detect the expression of CD69 and IFN-γby γδT cells, re-spectively.Results The γδT cells derived from healthy subjects showed cytotoxicity against the U2OS/MTX cells.Moreover, stronger cytotoxic activities of γδT cells were detected after pretreatment of U2OS/MTX cells with ZOL (1 μmol/L) for 24 hours (P<0.01).Stimulation with U2OS or U2OS/MTX cells in-creased the expression of CD69 onγδT cells and enhanced the secretion of IFN-γbyγδT cells (P<0.05). Increased expression of CD69 and enhanced secretion of IFN-γwere induced in γδT cells in response to stimulation with ZOL-pretreated U2OS/MTX cells (P<0.01).Conclusion TheγδT cells showed cytotox-icity against U2OS/MTX cells.Pretreatment of U2OS/MTX cells with ZOL could enhance the cytotoxic ac-tivity of γδT cells.

Objective To investigate the role of epithelial cell apoptosis in the stress-related changes of intestinal mucosa barrier following traumatic brain injury. Methods Sixty-four health male Wistar rats were divided randomly into two groups: (1) traumatic brain injury model group (n =32) ,in which rats suffered from traumatic brain injury by Feeney's method; (2) control group ( n = 32) , rats suffered from sham operation. Each group were divided into four subgroups according 6 h, 12 h,24 h, and 48 h after operation ( n = 8, for each subgroup ). Ileum tissue were taken to observed the damage of the intestinal mucosa under microscope and electronmicroscope. The early apoptosis rate of intestinal mucosal cells were analyzed with Annexin Ⅴ-PI double stained and detected by flow cytometry. Results The intestinal mucosa were damaged and the intercellular space of intestinal mucosal were found increased in traumatic brain injury group. The early apoptosis rate of intestinal mucosal epithelial cells was significant increased in traumatic brain injury group than that in control group [6 h: (13.5 ±3.7)% vs (6.1 ± 1.7)%, P＜0.05; 12 h:(66.1±6.0)% vs (5.2±1.1)%, P＜0.05; 24 h:(39.8±4.8)% vs (8.4±2.6)%, P＜0.05;48 h: (7.5 ±1.3)% vs (6.6 ±0.5)%]. Conclusion The increased early apoptosis rate of intestinal mucosal epithelial cells might contribute to the stress-damage of the intestinal mucosa barrier in early stage of traumatic brain injury.

Objective To study the role of bile reflux in the pathogenesis of stress ulcer after acute brain trauma in rat. Methods The animal model was reproduced by Allen's method with modification. The bile acid concentration in the stomach and blood, the pH value of the gastsic content, and ulcer index (UI) were assayed 1, 3, 6, 24 hour after the stress, and they were compared with that of sham injury group. Results In stress group, compared with sham injury group, the amount of bile acid in the stomach was notably increased( P

Objective:To investigate the effect of typeⅠIFN on the cytotoxic activity ofγδT cells from peripheral blood mono-nuclear cells ( PBMCs) of healthy donors and osteosarcoma patients stimulated by zoledronate ( Zol) and IL-2 against OS.Methods:PBMCs from healthy donors and osteosarcoma patients were stimulated with Zol+IL-2 or Zol+IL-2+typeⅠIFN,respectively.After 14 days of culture,ex vivo expandedγδT cells were assessed by flow cytometry.γδT cell cytotoxicity against target cells was analyzed using a standard lactate dehydrogenase release assay.IFN-γsecreted fromγδT cells was determined by ELISA kits.Results:γδT cells from PBMCs of healthy donors and patients with OS were selectively expanded by Zol+IL-2 or Zol+IL-2+typeⅠIFN in vitro,respectively, and showed cytotoxicity against OS.In addition,γδT cells pre-treated by TypeⅠIFN showed significantly higher cytotoxicity against OS cells.Conclusion:Type I IFN can enhance humanγδT cells’ cytotoxic activity against OS.

Objective To investigate the feasibility of high resolution melting(HRM)curve analysis technique for detecting β-thalassemia gene mutations.Methods The HRM method was used to detect five kinds of common β-thalassemia mutations (-28, CD17,CD41-42,CD71-72 and IVS-2-654)in Guangdong province.Sixty specimens of the patients with suspectedβ-thalassemia were performed the HRM analysis.The results of HRM analysis were confirmed by the direct DNA sequencing.Results Among 60 specimens of the patients with suspectedβ-thalassemia,12 cases of heterozygosis mutant gene were detected,including 3 cases of-28,2 cases of CD17,5 cases of CD41-42,2 cases of CD71-72 and 2 cases of IVS-2-654 gene mutation;2 cases of homozygosis mu-tant gene were detected,including 1 case of-28 and 1 case of CD41-42 homozygous mutation (both were amniotic fluid specimens). The results of HRM analysis were consistent with the DNA sequencing results.Conclusion The HRM method can accurately de-tect five kinds of common β-thalassemia mutations in Guangdong province,has the advantages of simpleness,rapidness and high sensitivity and is expected to be a new method for screening β-thalassemia mutation in clinic.

BACKGROUND:At present, the copper-bearing intrauterine device, a kind of class III medical devices, is
commonly used in China. However, there is no clear conclusion about whether it has impact on the embryo or
fetus in some cases, such as unexpected pregnancy during long-term implantation and pregnancy in a short time after removing it.
OBJECTIVE: To evaluate the safety of copper-bearing intrauterine device by observing the influence of
copper-bearing intrauterine device extracts on pregnant rats and rat fetuses by tail vein injection in the sensitive period of teratogenesis.
METHODS: A total of 60 fertilized rats were divided into control group, high dosage group, middle dosage group, and low dosage group. The copper-bearing intrauterine device extracts were prepared by the continuous
extraction method. Different concentrations (0.2, 0.1, 0.05 g/mL) of copper-bearing intrauterine device extracts were injected by the tail vein at the 1st day of pregnancy in the latter three groups at a dosage of 0.01 mL/g per day. The control group was given the same amount of normal saline. The injection lasted for 20 days. Then, the pregnant rats were sacrificed to measure body mass, check both sides of the uterus and internal organs, isolate fetal rats, as wel as record the quality of uterus and fetal rats, corpus luteum, implantation numbers, the number of
stilbirths, then number of live births and the number of fetal absorption. The fetal rats were determined in the folowing aspects: body mass, body height, tail length, the ossification degree and appearance of the occipital bone, bone and visceral anomalies.
RESULTS AND CONCLUSION: The number of births, implantation numbers, the number of live births, the number of
corpus luteum, the percentages of live births and stilbirths, the number of resorbed fetuses, and the weight of uterus and fetal rats in the control group showed no difference from those in the other three groups (P > 0.05). No malformation in the internal organs occurred. Compared with the control group, the high, middle and low dosage groups showed no
difference in the height, tail length, body mass, and ossification degree of the occipital bone of fetal rats (P > 0.05). No malformation in the appearance, skeleton and internal organs occurred in the fetal rats. These findings indicate that there were no maternal toxicity, abnormal embryonic growth or rat fetus anomalies after injecting copper-bearing intrauterine device extracts into pregnant rats in sensitive period of teratogenesis.