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.

OBJECTIVE: To investigate the relationship between the expression of vascular endothelial growth factor C (VEGF-C) and angiogenesis and lymphangiogenesis in papillary thyroid carcinoma (PTC). METHODS: Seventy-two PTC cases were divided into 3 groups according to the level of invasion: papillary microcarcinoma group (PMC group), intrathyroid carcinoma group (IPC group), and extrathyroid carcinoma group (EPC group). They were again divided into 2 groups according to lymph node metastasis: lymph node metastasis group and lymph node no-metastasis group. The expressions of VEGF-C, CD105 and vascular endothelial growth factor receptor-3 (VEGFR-3) were detected by SP method of immunohistochemical staining. The expression of VEGF-C was analyzed quantitatively by image analysis system, and the PI of VEGF-C (VEGF-C-PI), the number of MVD (microvessel density), and LVD (lymphaticvessel density) were obtained. RESULTS: The VEGF-C-PI of lymph node metastasis group (23.15 +/- 3.75) was higher than that of lymph node non-metastasis group (14.54 +/- 2.93) (P <0.01). MVD was 35.25 +/- 2.06 in the PMC group, 41.75 +/- 5.46 in the IPC group, and 52.58 +/- 4.16 in the EPC group, which showed the elevatory tendency with the increase of invasion (P < 00.5). LVD was 6.00 +/- 0.81 in the PMC group, 13.80 +/- 1.81 in the IPC group, and 19.17 +/- 2.96 in the EPC group, which again showed the elevatory tendency with the increase of invasion (P <0.05). The LVD of lymph node metastasis group (19.56 +/- 2.45) was significantly higher than that of lymph node non-metastasis group (12.48 +/- 2.84) (P < 0.05). VEGF-C was positively correlated with MVD and LVD (r = 0.743, 0.90, P <0.01). CONCLUSION The expressions of VEGF-C and LVD are related to lymph node metastasis of PTC. MVD and LVD are related to the invasion of PTC. VEGF-C may play an important role in the angiogenesis and lymphangiogenesis.
Adenocarcinoma, Papillary ; blood supply ; metabolism ; pathology ; Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Lymphangiogenesis ; Male ; Middle Aged ; Neovascularization, Pathologic ; metabolism ; Thyroid Neoplasms ; blood supply ; metabolism ; pathology ; Vascular Endothelial Growth Factor C ; biosynthesis ; genetics

Calcinosis ; complications ; Decalcification Technique ; Humans ; Meningeal Neoplasms ; complications ; Meningioma ; complications ; Specimen Handling ; methods ; Ultrasonics

OBJECTIVETo investigate and clarify the key surgical techniques and evaluate clinical effectiveness of modular cementless femoral stems(LINK MP modular stem) in complex revision hip arthroplasty.

METHODSFrom January 2002 to December 2013, 470 patients in Chinese PLA General Hospital received revision hip arthroplasties using MP stems. Among them, 246 patients were followed for an average of 5.7 years (ranged, 0.5 to 11 years). There were 148 males and 98 females, with an average age of 62.5 years old (ranged from 26 to 83 years old). The most common reason for revision was aseptic loosening of femoral stems (107 cases, 43.5%), followed by prosthetic joint infection (104 cases, 42.3%). Other reasons included unexplained hip or thigh pain, periprosthetic femoral fracture, prosthesis wear and dislocation. According to Paprosky classification of femoral deficiency, there were 171 cases of type IIIA (69.5%), 14 cases of type I, 39 cases of type II, and 22 cases of type IIIB. The average preoperative Harris hip score (HHS) was 37.4 ± 7.5. In addition, radiographic presentation, degree of patients' subjective satisfaction, and clinical effectiveness were assessed as well.

RESULTSThe average Harris score was im- proved to 87.8 ± 4.2 after operation,with significant statistical difference when compared to preoperative score (t = 92.13,P = 0.00). Analysis of patients' subjective satisfaction for leg length discrepancy, stability and overall degree of satisfaction showed that the number of patients with great satisfaction were 27, 60, 61 respectively. The most common complication was the periprosthetic fracture (intraoperatively 21 cases and postoperatively 8 cases). Other complications included infection (7 cases), dislocation (5 cases), numbness possibly caused by incomplete sciatic nerve injury (6 cases) and thigh swelling (3 cases). The X-ray taken at the final follow-up revealed that prosthesis subsidence occurred in 4 cases, 2 of which needed a second revision.

CONCLUSIONThe application of modular cementless femoral stems (LINK MP modular stem) in complex revision hip arthroplasty (especially in cases with the serious defect of proximal femoral bone) shows good results at both the short-term and long-term follow-up, especially in the revision for prosthetic infection with the high success rate. In order to avoid complications and improve clinical outcomes,the surgeons should carefully select proper surgery candidates,perform the procedure with care, and master the key surgical techniques.

Adult ; Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; adverse effects ; methods ; Female ; Hip Prosthesis ; Humans ; Male ; Middle Aged ; Patient Satisfaction ; Postoperative Complications ; therapy

The purity of 4,4′-dimethoxy-5,6,5′,6′-bis (methylenedioxy)-2′-morpholine methylenebiphenyl-2-methyl formate methanesulfonate (IMH), a new drug for fatty liver treatment, was determined through differential scanning calorimetry (DSC). Analysis of two-factor non repeatability method was performed in the investigation the effects of two factors (heating rate and sample weight) on purity determination. The DSC experimental parameters were optimized as follows: heating rate was 10 ℃·min^-1, temperature range was 150-300 ℃, sample weight was 2.0-4.1 mg, and N₂ flow rate was 80 mL·min^-1. The linear correlation coefficient (r) of this DSC method was 0.999 8. The RSD value (n = 6) of precision was 0.03%. The standard value and uncertainty of the purity results of the multiple batches of IMH drugs were (99.74 ± 0.29)%, (99.91 ± 0.28)%, (99.90 ± 0.28)%, and (99.81 ± 0.28)% with inclusion factor (K) of 2 and confidence probability (P) of 0.95. The results were basically consistent with the results of the mass balance method. The DSC mehod is a simple, rapid and accurate method, and provides a new reference method for determining the purity of IMH drugs, improves the accuracy and reliability of purity determination.

Objective To observe the distribution and metabolism of arsenic speciation in urine of rats exposed to different concentrations of dimethylaraenic acid (DMA) through drinking water.Methods Thrity six weaning Wistar rats were randomly divided into normal control,low-dose group and high-dose group,12 rats in each group(6 female and 6 male); average body weight of female rats was (60 ± 5)g,and male rats was (50 ± 5)g.All rats of the 3 groups were given DMA at concentrations of 0,100,200 mg/L,respectively,corresponding to their specific groups through drinking water for 10 weeks.Inorganic arsenic(iAs),monomethylarsenic acid(MMA),DMA and trimethylarsenic compound (TMA) in urine were measured by hydride generation trapping and ultrahypothermia coupled with atomic absorption spectrometry.Results After feeding for 10 weeks,the differences of rat urinary concentrations of iAs,MMA,DMA and TMA between normal control,low-dose group and high-dose group were statistically significant(x2 =25.441,25.942,25.751,17.767,all P＜ 0.01).Urinary concentrations of iAs,MMA and DMA(2.541,4.383,24.447 mg/L) of low-dose group were significant higher than those of normal control (0.784,0.000,0.743 mg/L,all P ＜ 0.05) ; iAs,MMA,DMA and TMA(3.978,7.186,35.112,4.518 mg/L) of high-dose group were significantly higher than those of normal control(0.784,0.000,0.743,0.000 mg/L,all P ＜ 0.05).The concentrations increased along with increasing doses of DMA concentrations in drinking water(all P ＜ 0.05).Conclusions After rats are exposed to DMA,most of the DMA are excreted in unchanged form in urine and a small portion of DMA is metabolized into TMA.

AlM:To discuss the protective effect ofα-mangostin on retinal light damage in mice.METHODS:Totally 30 Balb/c mice, aged 6~8wk, were randomly divided into the control group, light-exposure group and α-mangostin group. Every group contained 10 mice. Mice of α-mangostin group were treated with alpha-mangostin at the dose of 30mg/( kg · d ) body weight by intragastric administration daily for 7d, and then exposed to white light at the 5th d. The light-exposure group and α-mangostin group were exposed to 5 000 ± 200lx white light-emmiting diodes (LEDs) for continuously 1h to establish the mice model of retinal light damage. Flash -electroretinograme was recorded 72h after light exposure. The changes in retinal morphology of mice were observed by light microscopy. Retinas were extracted to detect the malondialdhyde ( MDA ) content change of the retinal homogenate.RESULTS: Flash-electroretinogram ( F-ERG ) showed that retinal dysfunction was less severe in α-mangostin group than in light-exposure group ( P<0. 05 ). Light microscopy test showed that retina structural damage was less severe in α-mangostin group than in light-exposure group (P<0. 05). The level of MDA in retinal tissue of α-mangostin group was significantly lower when compared with light-exposure group (P<0. 05).CONCLUSlON: α-mangostin inhibits lipid peroxidation induced by light damage and protect retina against light damage.

Twin pregnancy with mosaic partial hydatidiform mole (PHM) and survival of two healthy fetuses following in vitro fertilization and embryos transfer (IVF-ET) is a rare situation and is considered a challenge for management. A 32-year-old Chinese woman conceived twin pregnancy following IVF-ET. At 22 weeks' gestation, an additional intrauterine echogenic mass with features of PHM were shown by successive ultrasound examinations. At 35 weeks' gestation, two live male infants and two placentas were delivered by caesarean section (CS). Histologic examination of the abnormal placenta confirmed mosaic PHM. Genetic study showed the abnormal placental mosaicism (expressed in molar-69XXY and normal vili-46XY), co-existing with a hypospadia new-born (46XY) in one amniotic sac. However, the other one was normal. Serial serum β-hCG levels showed a declining trend and serum β-human chorionic gonadotropin (hCG) were undetectable at 6 months after delivery. The case demonstrated that it is possible to prolonged gestation by PHM under close surveillance during the entire pregnancy.
Adult ; Female ; Fertilization in Vitro ; Humans ; Hydatidiform Mole ; genetics ; Infant, Newborn ; Male ; Mosaicism ; Placenta ; metabolism ; pathology ; Pregnancy ; Pregnancy, Twin ; genetics

Three tricyclic [6,5,7] and six tetracyclic [6,5,5,5] novel indole alkaloids were synthesized and evaluated on triglyceride inhibitory activities for the first time. Among them, compound 4c showed the most potent activity with IC₅₀ value of 6.35 μmol·L^-1. Meanwhile, compound 4c also exhibited a good safety profile at the cellular level. Preliminary mechanism study indicated that 4c might increase intracellular lipid metabolism by activating AMPK. These results provide a novel family of lead compounds for the discovery of anti-NAFLD candidates.

OBJECTIVETo study the effect of transforming growth factor-β activated kinase-1 (TAK1) gene silencing on the proliferation and apoptosis of Kasumi-1 cells induced by arsenic trioxide (As₂O₃).

METHODSAcute myeloid leukemia with t(8;21) cell line Kasumi-1 cells were treated with As₂O₃ or in combination with TAK1 siRNA interference technology. The experiment was divided into four groups: Kasumi-1 cells without any treatment, TAK1 specific siRNA transfection alone, Kasumi-1 cells treated with different concentration of As₂O₃, TAK1siRNA transfection combined with As₂O₃. CCK-8 was used to detect the cell viability. The expression of phosphorylated c-Jun N-terminal kinase (P-JNK) was determined by Western Blot. Cell apoptosis and growth were examined by morphological and colony formation assay.

RESULTSAfter Kasumi-1 cells were treated with As₂O₃, the rate of cell inhibition was concentration-dependent, and the 50% inhibitory concentration was 3.5 μmol/L. The highest expression level of P-JNK appeared in 30 minutes after cells were treated with As₂O₃. The apoptosis rates of Kasumi-1 cells without any treatment, TAK1 siRNA interference alone group, As₂O₃ alone group and the combined group were (5.02 ± 1.13)%, (6.18 ± 0.28)%, (48.33 ± 2.70)% and (86.07 ± 2.21)%; colony formation rates were (73.83 ± 2.78)%, (76.03 ± 1.46)%, (55.07 ± 1.50)% and (22.20 ± 1.15)%; apoptosis rate of TAK1 siRNA group and the untreated group has no significant difference (P = 0.052); colony formation rate between TAk1 siRNA group and the untreated group has no significant difference (P = 0.179), but the difference in other groups was significant (P = 0.000).

CONCLUSIONSilencing the expression of TAK1 can enhance the anti-proliferative and pro-apoptotic effect of As₂O₃ on Kasumi-1 cells, and its mechanism may be through the TAK1 downstream JNK signal pathway.

Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Cell Line, Tumor ; Humans ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Leukemia, Myeloid, Acute ; enzymology ; pathology ; MAP Kinase Kinase Kinases ; genetics ; metabolism ; Oxides ; pharmacology ; RNA Interference ; RNA, Small Interfering ; genetics ; Signal Transduction