Pichia pastoris is a new eukaryotic expression system with great potentials, has unexampled advantages in expressing the heterologous proteins and has gotten more and more wide application. The following aspects were described in detail: the advantages of Pichia pastoris in the expression of heterologous genes, the mechanism of the heterologous gene integrations, the glycosylation of the expressed protein and the modification of the post-translated protein.

·AIM: To evaluate the function of primary human corneal endothelial cells(HCEC) serving as immunological cells. ·METHODS: Expression of HLA-DP, -DQ, -DR, CD40, CD80, and CD86 was determined by immunohistochemical methods. Meanwhile, purified peripheral blood mononuclear cells(PBMC) were cocultured with primary HCEC which were pre-treated with and without γ-IFN respectively; activation of lymphocytes was determined by FACS analysis.·RESULTS: In coculture system, T lymphocyte was activated by primary HCEC, HLA-DP, -DQ, -DR and CD40 expression were increased by γ-IFN induction. Costimulatory molecular CD80 was shown on the endothelial cells. ·CONCLUSION: Primary HCEC were assumed to be involved in the corneal transplantation rejection process as potential antigen presenting cells(APC).

Objective To analyze re-treatments of recurrence after the pelvic floor repair surgery.Methods The protocol and the effect of re-treatments were investigated by reviewing and analyzing the clinical data of 81 recurrent patients (grade Ⅱ and above),who had received the pelvic floor repair surgery from January 2011 to January 2016.Pelvic organ prolapse quantitation system (POP-Q) and two questionnaires about quality of life [pelvic floor distress inventory-short form 20 (PFDI-20) and pelvic floor impact questionnaire short form (PFIQ-7)] were used to evaluate objective and subjective efficacy,respectively.Results Among 81 recurrent patients who were followed up for a median of 35 months (10-69 months),78 cases (with prolapse up to grade Ⅲ or Ⅳ) were treated by surgical operation with both objective cure rate and subjective satisfaction being 100％ (78/78);3 cases (with grade Ⅱ prolapse) were treated by pelvic floor electrical stimulation biofeedback,and 1 case among the three cases had the vaginal foreign body sensation,the subjective satisfaction was 2/3.The methods of surgical operation for the 78 recurrent patients included:total pelvic floor reconstructive surgery (55 cases;3 of which involve trachelectomy),anterior pelvic reconstructive surgery (2 cases),posterior pelvic reconstructive surgery (3 cases),Y-mesh sacral colpopexy (2 cases),colpocleisis (11 cases),vaginal hysterectomy combined posterior fornix forming (3 cases),and vaginal hysterectomy combined posterior pelvic reconstructive surgery (2 cases).Conclusion The extent of recurrence,the recurrent site and complications must be carefully considered and evaluated for re-treatments of recurrence after pelvic floor repair surgery,and then an appropriately individualized re-treatment protocol could be designed for each of the patients.

Objective To investigate the role of nuclear factor-kappa B(NF-?B) and monocyte chemoattractant protein-1(MCP-1) in the pathogenesis of uric acid nephropathy.Methods Twenty male SD rats were divided into control group(group C) and model group(group M).The model rats with uric acid nephropathy were made using adenine.Immunohistochemistry was used to detect the expressions of NF-?B and MCP-1 in 18 days and the serum uric acid and renal fuction were measured respectively.Results In comparison with group C,the expressions of NF-?B and MCP-1 in the rats of group M were significantly increased(all P

Background Chemical crosslinking agent can be used to strengthen the intensity of sclera tissue,but the intensity of the sclera may be influenced by different crosslinking methods.Objective The aim of this study was to compare the effectiveness of collagen crosslinking on porcine sclera between whole-eye crosslinking method and scleral strip crosslinking method.Methods Whole-eye crosslinking or sclera strip crosslinking was performed on 70 fresh porcine eyeballs in five groups using 1％ genipin,1％ glutaraldehyde or PBS respectively for 40 minutes.After crosslinking,10 sclera strips with l0 mm×4 mm from the temporal lateral were prepared in every group for the stress-strain measurement using a Instron5848 microtester,and the other 4 scleral strips in each group were extracted for the thermal shrinkage temperature test.Results Biomechanical property test reveled that the elastic modulus value of sclera strips reduced by 70.0％-82.8％ in the whole-eye crosslinking method group compared with scleral stip crosslinking method group after treated with 1％ genipin ((8.98 ± 1.81) MPa vs.(10.85 ± 1.83) MPa,t =3.375,P =0.003)) and 1％ glutaraldehyde((12.78 ±2.91) MPa vs.(18.25 ±5.16) MPa,t =4.007,P =0.001)) ;The tensile stress of whole-eye crosslinking method group was 54.9％-90.1％ of scleral stips method group,showing significant decline after corsslinked of whole-eye in 5％,10％,15％ and 20％ strain conditions (all P ＜ 0.05).Thermomechanics test showed that the thermal shrinkage temperature was lower in the whole-eye crosslinking group compared with scleral stip crosslinking group after treated with both 1％ genipin ((68.8 ±0.9)℃ vs.(74.8± 1.3)℃,t=11.129,P=0.000)) and 1％ glutaraldehyde((73.3±0.9)℃ vs.(79.3±1.3)℃,t=11.112,P=0.000)).Conclusions Different crosslinking methods have an influence on the efficacy of collagen crosslinking on porcine sclera.Sclera strip crosslinking offers a better crosslinking intensity for selera.

Objective：To evaluate bioequivalence and relative bioavailability of domestic and imported repaglinide tablets in healthy volunteers. Methods： Twenty two healthy male volunteers were randomized into A and B groups. A single dose (4 mg) of domestic and imported repaglinide tablets were given respectively according to an open 2-way crossover study design. The washout period was 1 week. Plasma concentrations of repaglinide were determined by HPLC method. Results：The pharmacokinetic parameters of domestic and imported drugs were as follows: t1/2 were(0.86±0.24) and (0.83±0.31) h；tmax were( 0.79±0.37) and (0.75±0.41) h；cmax were (52.43±20.92) and (53.32±24.94) μg/L. AUC0-t were (79.87±36.48) and (74.95±30.57) μg*h*L-1，respectively. The relative bioavailability of domestic formulation was (106.55±16.15)%. Conclusion： The results of variance analysis and two one-side t test show that 2 formulations are of bioequivalence.

Objective To review the surgical managements of patients with traumatic false aneu- rysms in the extremities.Methods From January 1990 to April 2006,17 patients with traumatic false aneurysms in the extremities were admitted into our hospital.Fourteen patients were treated by vascular repair including vascular repair in seven cases,end to end anastomosis in one,synthetic grafting in one, autogenous vein grafting in one,and direct ligation in four.Three patients were treated nonoperatively, but with local compressive dressing.Results There were no deaths or gangrenes in all cases.The clinical manifestations vanished after the treatment.The mean follow-up period was 13.2 months.The function of the injured extremities recovered satisfactorily.Conclusion Different types of traumatic false aneurysms should be managed by different therapeutic procedures after the diagnoses is made.

Objectives To detect the cord blood vitamin D level in neonates and to determine the association between the cord blood vitamin D level and neonatal outcomes. Methods A total of 223 eligible mother-and-singleton-offspring pairs were recruited. The information of mothers' pregnancy was collected by questionnaires. The weight, length, and head circumference of neonates were measured. The levels of 25(OH)D in cord blood of neonates and in blood of late pregnancy mothers were determined by chemiluminescence immunoassay. Results The median concentration of 25(OH)D in cord blood was 20.7 nmol/L, and 82.1% of neonate had vitamin D deficiency, and 12.1% had severe vitamin D deficiency (<10 nmol/L). The concentration of 25(OH)D in cord blood was consistent with that in blood of late pregnancy mother. The distribution of concentration of 25(OH)D in cord blood was significantly different in neonates in different seasons of birth (P<0.05). There were more cases <10 nmol/L in winter and spring. The concentration of 25(OH)D in cord blood had no significant associations with the incidences of low birth weight (LBW) and small for gestational age (SGA) (P>0.05). After the variables of sex, gestational age and birth season are controlled, the birth weight and head circumference were significantly different in neonates with different concentrations of 25(OH)D in cord blood (P<0.05). Conclusions The concentration of 25(OH)D in cord blood in term neonates was generally lower. The vitamin D status in neonates was consistet with that in their late pregnancy mothers. Cord blood 25(OH) D levels were associated with neonates' birth weight and head circumference, but it should be confirmed by larger sample size in the future.

Objective:To analyze the result of smooth pursuit test(SPT) in unilateral vestibular peripheral vertigo and investigate its influencing factors.Method:Smooth pursuit test (SPT) and spontaneous nystagmus (SN)were examined in one hundred and eighty-five patients with unilateral peripheral vertigo(case group) and 51 normal persons(control group) by Video-Nystagmography (Synapsys, France), and the gain of SPT and SN were selected as the observation parameters in order to analyze the waveform and gain of SPT and the relativity between SN and the gain of SPT.Result:Of the 185 patients, 105(56.8%),72(38.9%) and 8(4.3%) cases producedⅠ,Ⅱ and Ⅲ waveforms respectively. Of these patients, 58(31.4%) demonstrated SN and none had Ⅳ waveform. While of 51 normal persons, 38(74.5%), 13(25.5%) persons producedⅠand Ⅱwaveforms repectively and there were no Ⅲ, Ⅳ waveforms or SN. There was statistical significance between the stong and weak gain of SPT in these two groups. Weak gain was significantly different between two groups. The stong and weak gain of SPT in case group were 0.86±0.06,0.80±0.06; 0.78±0.09, 0.65±0.1; 0.68±0.13, 0.45±0.12. The relativity between SN and the gain of SPT was positive when they had same direction(r_s=-0.63,P＜0.05)and negative when opposite (r_s=0.34,P＜0.05).Conclusion:Ⅰ,Ⅱ,Ⅲ three waveforms of SPT could appear in unilateral vestibular peripheral vertigo and the corresponding gains are gradually decreasing.SN is the influencing factor of SPT.

Objective To investigate the feasibility and safety of transvaginal endoscopic cholecystectomy.Methods The clinical data of 88 female patients who underwent cholecystectomy at the Qilu Hospital of Shandong University from May to November, 2009 were retrospectively analysed. Among all the patients, 32 received transvaginal endoscopic cholecystectomy ( NOTES group) and the remaining 56 patients received laparoscopic cholecystectomy (LC). Thirty-two patients who received LC at the same period were selected (LC group)acccording to age, body mass index, type and severity of disease to conduct a matched case-control study. The differences in time span of postoperative pain, anodyne dose, enterokinesia recovery time, operation time, out-ofbed activity time, average hospital stay and hospitalization expenses between the two groups were compared using the paired t test. Results Cholecystectomies were successfully carried out for all the patients. The intraoperative blood loss, operation time, degree of pain, anodyne doses, enterokinesia recovery time, out-of-bed activity time,average hospital stay and hospitalization expenses were (5.7 ± 1.5 ) ml, ( 76 ± 27 ) minutes, 2.2 ± 0.6, ( 10 ±6) mg, (25±5) hours, (9±3) hours, (2.1 ±1.2) days and (1.12±0.34) ×104 yuan in NOTES group, and they were ( 13.9 ± 3.1 ) ml, (38 ± 16) minutes, 6.7 ± 1.5, (28 ± 8) mg, (45 ± 8) hours, (26 ± 6) hours,(4.3 ± 2.1 ) days and ( 1.54 ± 0.18 ) × 104 yuan in the LC group. There were significant differences between the two groups (t = 5.098, - 4.712, 2.417, 3.203, 3.089, 4.136, 4.786, 3.917, P ＜ 0.05 ). Conclusion Transvaginal endoscopic cholecystectomy is safe and feasible, and it is superior to tranditional LC.