Objective To study the implication of phosphoinositide-specific phospholipase C?2(PLC-?2) in gastric cancer MGC80-3 cells treated by TPA.Methods The effect of 12-O-tetradecanoylphorbol-1,3-acetate(TPA) on gastric cancer MGC80-3 cells was detected and analyzed by DAPI staining under fluorescence microscope.The effect of TPA on the expression level of PLC-?2 protein was detected by Western blotting,when nuclear and cytoplasmic protein fractions were prepared through lysis of cell and centrifugation.Localization and translocation of PLC-?2 were observed under laser scanning confocal microscope with immune-fluorescence technique.After MGC80-3 cells were pretreated by U73122(PLC-?2 inhibitor),the effect of TPA on PLC-?2 was detected by Western blotting and laser scanning confocal microscope with immunefluorescence technique,and the effect of U73122 on MGC80-3 cells followed by TPA was observed and analyzed by DAPI staining under fluorescence microscope. Results TPA induced the apoptosis of gastric cancer MGC80-3 cells.Meanwhile,TPA increased the expression level of PLC-?2 protein and induced its translocation from cytoplasm to nucleus.U73122 inhibited the effect of TPA on the expression level of PLC-?2 protein,but it did not affect on the cell apoptosis induced by TPA.However,the translocation of PLC-?2 protein induced by TPA was not inhibited by its inhibitor(U73122).Conclusion Although TPA could increase the expression level of PLC-?2 protein,it is not directly related to the cell apoptosis induced by TPA.However,its translocation might be related with the apoptosis of gastric cancer.

Objective To study the expression of protein kinase B (Akt/PKB) isoforms in the gastrocnemius muscle of rats at different ages. Methods The expression levels of mRNA and protein of three Akt isoforms in the gastrocnemius muscle of 30-month-old and 6-month-old rats were detected respectively by RT-PCR and Western blotting. Data were analyzed statistically. Results (1) The levels of Akt1 and Akt2 mRNA in the 30-month-old rats was significantly higher than those in the 6-month-old rats (t=7.124, P=0.000; t=2.598, P=0.021), however, no significant difference was found in the level of Akt3 mRNA between the two groups (t=0.460, P=0.653) . (2) The level of Akt-Thr308 phosphorylation in the 30-month-old rats was significantly lower than that in the 6-month-old rats (t=-9.861, P=0.000), while the level of Akt2 protein in the 30-month group was significantly higher than that in the younger rats (t=7.522, P=0.000). No significant differences were detected in the levels of Akt1 and Akt3 proteins between the two groups (t=0.469, P=0.646; t=0.058, P=0.955). Conclusion The expression levels of three Akt isoforms in the gastrocnemius muscle of rats change with age, suggesting that the three isoforms of Akt have different functions in the gastrocnemius muscle metabolism of rats at different ages.

Objective To investigate the expression and role of Caspase-3 during the development of non-traumatic avascular/aseptic necrosis of femoral head(ANFH).Methods Tissue samples were obtained from 22 femoral heads of 21 patients with ANFH.The samples were divided into three groups:early stage(6 cases),middle stage(8 cases) and late stage(8 cases) groups according to the progress of the disease.Meanwhile,19 tissue samples of 19 femoral heads were obtained from 19 cases of acute femoral neck fracture and set as a control group.The level of apoptosis of each group was compared in terms of the percentages of TUNEL positive osteocytes and empty osteocyte lacuna,and then the activity of Caspase-3 was detected by colorimetric method.Results Both the pecentages of empty osteocyte lacunae(F=45.43,P=0.000) and the TUNEL positive osteocytes(F=120.86,P=0.000) increased significantly with the progress of ANFH.The two indexes in the early and middle stage groups were significantly higher than those in the control(q=18.899,P0.05).Caspase-3 activity was not significantly related to the capacity of osteogenesis(r=0.126,P=0.425).Conclusions Caspase-3 is not the major executor of apoptosis in ANFH.It is very likely that it does not play a nonapoptotic role in the progress of the diseases.

Objective To probe into the remodeling and maturity process along with extension of time in anterior cruciate ligament (ACL) reconstruction with hamstring autograft in order to confirm the date of maturity postoperatively. Methods Thirty-three patients after arthroscopic ACL reconstruction with hamstring autograft, were undergone second-look arthroscopical surgery. Meantime, biopsy specimens were obtained from the mid-zone of the hamstring graft. As a control group, specimens of normal ACL were obtained from total knee replacement of four cases, and specimens of semitendinous tendons obtained from ACL reconstruction of the four cases. The interval from initial reconstruction to second-look arthroscopy ranged from 2 to 36 months(mean 11.9 months). Patients were divided into different groups by postoperative time such as 1-, 4-, 7-, 10-, 13-, 18- and 25- month group. Thirty-three specimens were ordinarily sliced up into longitudinal sections. The sections were stained with hematoxylin and eosin so that cell and collagen fibrils were observed under light microscope, then compared the results with those of control group. Results Under microscopical observation, it was showed that tissue of hamstring graft was remodeled from more blood vessels and cellular amount to normal one, and from an irregular orientated crimp pattern of collagen to regular orientated crimp pattern of collagen in a time-dependent manner. Decreasing cellular amount, changing cell type, and a regular orientated crimp pattern of collagen of the hamstring graft were observed in 7- month group primarily, comparing with the original ACL. However, there existed a slower process in the other groups that are older than that of 7- month. Conclusion There are well histological features in the process of remodeling and maturation of the hamstring autograft after operation following extension of time, such as better survival of early period, and remodel of vessels and collagen. The process of remodeling and maturation in the hamstring autograft is similar to BPB autograft. The maturation period of the hamstring autograft after ACL reconstruction in human patients appears between 7 month and 9 month postoperatively.

AIM: To discuss the safety of hands chopping phacoemulsification in patients with microcoria cataract.METHODS:Hands chopping phacoemulsification with intraocular lens implantation was used for the microcoria cataract of 30 patients ( 32 eyes ) . Their visual acuity, pupil, and complication were observed in postoperative 1d,1wk and 1mo.
RESULTS: Postoperative naked vision be or more than 0. 3 were in 27 eyes (84. 4%) at one day, be or more than 0. 3 were in 30 eyes (93. 8%) at one week, be or more than 0. 5 were in 28 eyes (87. 5%) at one month. All pupil returned to round or oval. No synechia happened in postoperative 1mo.
CONCLUSION:Hands chopping nucleus operation is safe and effective for uveitis combined with microcoria phacoemulsification.

Objective To evaluate the safety and efficacy of intra-arterial thrombolysis in patients with acute cerebral ischemic stroke.Methods Twenty-one patients with acute internal carotid circulation infraction(internal carotid 3,MCA 12,ACA 5,lenticulostriate in 1) were treated with intra-arterial thrombolysis of recombinant tissue plasminogen activator(rt-PA) which was performed within 2-6 hours of symptom onset.Recanalization was observed during the operation.Intracerebral hemorrhage(ICH) was monitored immediately and 24 h after the treatment by CT or MRI scanning.Chinese stroke scale was used to evaluate the recovery of neurological functions pre-operatively and 30 d after the treatment.Results All the 21 patients were 100% success in receiving intra-arterial thrombolysis technique and revealed 16 having the degree of recanalization of 2 to 3 grade as regards to TMI,16 patients' degree of recanalization reached 2 to 3 grade according to TMI;5 patients showed 1 to 2 grade.Symptomatic ICH was observed in 3 patients,with two dead.Arterial dissection was found in one patient.Thirty days after the operation,17 patients' cerebral function reduced over 50 percent;2 less than 50 percent;and 2 died.The patients achieved 2 to 3 grade of recanalization were obviously getting better than those achieving 0 to 1 grade.Conclusions It is adapt to have intra-arterial thrombolysis with six hours from onset;but still have the danger of severe ICH.The treatment should be started as early as possible.

ObjectiveTo study the correlation of urinary iodine and thyroid function in elderly men in Harbin,and to provide the basis for formulation of health measures for the elderly.MethodsSeventy five cases of clinically healthy elderly men were enrolled for check-up of urinary iodine,thyroid function and B-ultrasound in Geriatric Ward the Forth Affiliated Hospital of Harbin Medical University in 2010.The subjects of investigation were divided into iodine appropriate and iodine sufficient groups and thyroid function parameters and B-ultrasound results were compared.ResultsThe average age of the 75 cases of healthy elderly men was (79.07 ± 4.78) years old and the median of urinary iodine was 198.4 μg/L.There were 62.67％ (47/75) elderly males whose iodine nutritional status was appropriate,but there were still some individuals(6.67％,5/75) in the iodine excess state.The level of TSH of the iodine appropriate group [(1.91 ± 0.82)mU/L] was lower than that iodine sufficient group [(4.98 ±0.60)mU/L,t =12.58,P ＜ 0.05],while the level of FT3 of the iodine appropriate group[(4.71 ± 0.56)pmol/L]was higher than that iodine sufficient group[(3.31 ± 0.43)pmol/L,t =12.18,P ＜ 0.05].But the difference of FT4between the two groups [(14.91 ± 3.12),(14.06 ± 2.79)pmol/L] was not statistically significant (t =1.40,P ＞0.05].The thyroid volume of iodine sufficient group[(20.9 ± 6.1 )cm3] was higher than that iodine appropriate group [(17.9 ± 5.6)cm3,t =2.11,P ＜ 0.05].ConclusionsSufficient quantities of iodine intake may affect the thyroid of elderly people.Whether the quantity of iodine intake of the elderly population should be decreased or not need to be further studied.

OBJECTIVETo discuss the clinical effects and superiority of applying external fixation and bone transport to treat osteomyelitis and bone defect of femoral bone.

METHODSFrom August 2008 to December 2013,16 patients with osteomyelitis and bone defect of femoral bone were treated including 11 males and 5 females with an average age of 42 years old ranging from 13 to 62 years old. The average course of disease was 18 months ranging from 2 months to 4.5 years, and the average length of bone defect was 7.8 cm ranging from 4.5 to 15 cm. The bone defect of all cases were treated by external fixation and bone transport, the bone transport began at 1 week after operation, 1 mm per day and 4 times per day.

RESULTSAll patients were followed up for 10 to 36 months (means 22.5 months). One patient did not cooperate with treatment leads to the failure, then took the amputation. The remaining 15 cases of osteomyelitis were under control, including 12 cases of bone transport achieved one stage bone union, 3 cases achieved bone union via bone graft from iliac bone. The bone union time was 5 to 13 months(means 7.9 months). Thirteen patients almost obtained the same length of two lower extremities,2 patients had shortening of 1.5 to 2 cm. The time of moving the external fixation was from 6 to 16 months (means 9.3 months).

CONCLUSIONApplication of external fixation and bone transport is an effective method in treating the osteomyelitis and bone defect that can control the infection, eradicate wounds, and be the equalization of limb length.

Adolescent ; Adult ; Bone Transplantation ; External Fixators ; Female ; Femur ; surgery ; Humans ; Male ; Middle Aged ; Osteomyelitis ; surgery

BACKGROUND: Sinomenine is an alkaloid monomer extracted from a Chinese medicinal herb sinomenium acutum stem. It is used in the therapy of the rheumatoid arthritis and has clear and definite therapeutic effects, but the therapeutic mechanism is unclear.OBJECTIVE: To observe the effect of sinomenine at different doses in vitro on the activity of nuclear factor-κB(NF-κβ) and mRNA expressions of the tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β) andinterleukin-10 (IL-10) in the synovial cells of the rats with adjuvant arthritis(AA) to explore the probable mechanism of sinomenine in the treatment of rheumatoid arthritis(RA).DESIGN: A controlled repeated measuring study based on the cells.SETTING: Department of traditional chinese medicine and the institute of burn research of a military medical university.MATERIALS: This study was finished at the Laboratory of the Institute of Burn Research of Chinese PLA. The experimental animals were 25 healthy male Wistar rats of clean grade. The AA model rats were made and the synovial cells were collected and grouped as follows: normal control group, AA group,AA + sinomenine 30 mg/L group, AA + sinomenine 60 mg/L group, AA + sinomenine 120 mg/L group. The activity of the NF-κB was measured by the electrophoresis mobility shift assay(EMSA) . The mRNA expressions of the TNF-α, IL-1β and IL-10 were measured by reverse transcription-PCR assay.MAIN OUTCOME MEASURES: The results of the changes of the activity of the NF-κB and the mRNA expressions of the TNF-α, IL-1β and IL-10 in the synovial cells of the rats with adjuvant arthritis after the treatment with sinomenine at different doses.RESULTS: Compared with the normal control group, the activity of the NF-κB and the mRNA expressions of the TNF-α, IL-1β and IL-10 in the synovial cells in the AA group all increased significantly and the outcomes were 17±6, 0.570±0.047, 0.730±0.093, 0. 683 ±0.081 (t= 2.71 -4.07, P < 0.05). After the administration of sinomenine, the activity of NF-κB showed a good correlation with mRNA expressions of the TNF-αandIL-13(r=0.810, P <0.001; r=0.562, P <0.05), but no statistical relevance with mRNA expression of IL-10 was established. Sinomenine showed a dose-dependent inhibition on the activity of the NF-κB and the mRNA expressions of the TNF-α and IL-1β in a certain range of concentrations(30-120 mg/L), but no dose-dependent inhibition on mRNA expression of the IL-10 was observed.CONCLUSION: Through the inhibition of the activity of the NF-κB,sinomenine decreased the mRNA expressions of the TNF-α and the IL-1β in the synovial membrane cells.

Phage display technology has been developed as a powerful tool for selecting polypeptides or proteins with desired biological and physicochemical properties from huge random peptide libraries.Fragments of foreign peptides or proteins that are expressed as fused proteins displayed on the phage surface can keep their relatively independent spatial structure and biological activity,so that they can interact with their ligands to mimic selection of specific molecular epitopes,thus providing an efficient high-throughput screening system.Phage display has been used to allow rapid identification of peptide ligands for a variety of target molecules by an in vitro selection process called "panning".Phage display techniques can be widely exploited to construct tumor-associated antibody libraries,select polypeptides tumor-associated antigen,investigate antigen epitope and design vaccines and medicine,and are used especially as a tool for the diagnosis and treatment of tumor,gene therapy and tumor cell signal transduction research.Recent applications and advanced developments of phage display in cancer research are discussed in this article for the further reference to investigators.