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 examine the serum homocysteine,folic acid and Vitamin B12 levels in macrocytic anemia patients and observe their dynamic changes following therapy. Methods Homocysteine,folic acid and Vita-min B12 were analyzed by electro-chemiluminescence immunoassay. Complete blood cell count was analyzed by opti-cal method and resistance method. Results The homocysteine is significantly higher in nutritional megaloblastic a-nemia[ (71.26±27.84)μmoL/L ] than in drug-induced megaloblastic anemia[(11.44±5.06)μmol/L],in myel-odysplastic syndrome[ (9.51±4.13)μmol/L] and in the normal control group[(8.74±5.42)μmoL/L] (P<0.01). After treated with low-dosage folic acid and Vitamin B12,the patients with megaloblastic anemia presented slow declining but eventually normal homocysteine levels,compared with those received high-dosages. Conclusion Homocysteine can be used for differential diagnosis of macrocytic anemia. The duration of remaining of abnormal ho-mecysteine levels is related to the dosage of folic acid and Vitamin B12.

Air ; analysis ; Air Pollutants, Occupational ; analysis ; Chromatography, Gas ; methods ; Octanes ; analysis ; Workplace

OBJECTIVETo observe the effects of electroacupuncture (EA) with branch-foundation acupoint combination on the indices regarding pituitary-target gland axis in aging rats with yang deficiency, so as to explore its regulating mechanism on aging rats with yang deficiency.

METHODSForty healthy Sprague-Dawley female rats were randomly divided into a normal control group, a model group, an EA group and an EA control group, 10 rats in each group. Except the normal control group, the rats in the rest 3 groups were all treated with subcutaneous injection of D-galactose for 40 d, followed by intramuscular injection of hydrocortisone for 7 d to establish aging model with yang deficiency. After the successful establishment of model, the EA group was treated with EA at "Guanyuan" (CV 4), "Housanli" (ST 36) and "Baihui "(GV 20) while the EA control group was treated with EA at "Zhongji" (CV 3) "Yinlingquan" (SP 9) and "Yintang" (GV 29); the rats in the normal control group and model group were immobilized and fixed during the same time period. The treatments were given 6 times per week totally for 4 weeks. With radiation immunity analysis method, the 8 biological indices of pituitary-target gland axis, including thyroid-stimulating hormone (TSH), triiodothyronine (T3), tetraiodothyronine-4 (T4), adrenocorticotropic hormone (ACTH), corticosterone (CORT), estradiol (E2), luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were detected to observe the changes of their content.

RESULTSCompared with the normal control group, the serum level of TSH, T3, T4 and E2 were reduced in the model group (P<0.05, P< 0.01) while those of ACTH, CORT, FSH and LH were increased (P<0.05, P<0.01). Compared with the model group, the serum level of TSH, T3, T4 and E2 were increased in the EA group (P<0.05, P<0.01) while those of ACTH, CORT, FSH and LH were significantly reduced (P<0.05, P<0.01). Compared with the EA control group, the content of TSH was increased in the EA group without statistical significance (P>0.05), that of T3, T4 and E2 was increased (all P<0.05) and that of ACTH, CORT, FSH and LH was significantly reduced (all P<0.05).

CONCLUSIONThe electroacupuncture with branch-foundation acupoint combination has benign regulating effects on the key hormones of pituitary-target gland axis, which could effectively improve the dysfunction of pituitary-target gland axis that is caused by aging with yang deficiency; the efficacy of electroacupuncture with branch-foundation acupoint combination is superior to that of adjacent control acupoint combination.

Acupuncture Points ; Adrenocorticotropic Hormone ; blood ; secretion ; Aging ; blood ; metabolism ; Animals ; Disease Models, Animal ; Electroacupuncture ; Estradiol ; blood ; secretion ; Female ; Follicle Stimulating Hormone ; blood ; secretion ; Humans ; Pituitary Gland ; secretion ; Rats ; Rats, Sprague-Dawley ; Thyroid Hormones ; blood ; secretion ; Yang Deficiency ; blood ; metabolism ; therapy

BACKGROUND: The source of bone marrow mesenchymal stem cells (BMSCs) is limited, and the cellular morphology,proliferation and multi-directional differentiation capacities can vary during serial passages in BMSCs in vitro.OBJECTIVE: To study the effects of fibroblast growth factor (FGF) on cellular morphology, proliferation and differentiation of serially passaged BMSCs.METHODS: (1) BMSCs were isolated from Sprague-Dawley rats and cultured. These cells were passaged six times in vitro, and the cellular morphology was observed and photographed. (2) BMSCs at passage 6 were seeded into 96-well plates and randomly divided into control group and FGF treatment group. The proliferation of cells in both groups was detected with cell counting kit-8 kit at days 1, 2, 3, 4, 5, 6, 7 after culture. (3) BMSCs at passage 6 were seeded into 6-well plates and randomly divided into control group and FGF treatment group. After 7 days treatment with growth medium or growth medium containing FGF, the cellular morphology was observed and photographed. And then the cells of both groups were treated with osteogenic induction medium, adipogenic induction medium and chondrogenic induction medium for the next 7 days. The osteogenic, adipogenic and chondrogenic related genes (RUNX2, ALP, OCN;PPARγ2, AP2, ADIPOQ; SOX9, collagen II, aggrecan) were detected with real-time PCR. The protein expressions of RUNX2, PPARγ2, SOX9 were detected with western blot assay. (4) BMSCs at passage 6 were seeded into 6-well plates and randomly divided into control group and FGF treatment group. After 7 days treatment with growth medium or growth medium containing FGF, the cells were cultured with osteogenic induction medium, adipogenic induction medium and chondrogenic induction medium for the next 14 days. Then, alizarin red S staining, oil red O staining and alcian blue staining were performed.RESULTS AND CONCLUSION: (1) After in vitro passage for six times, the cellular morphology changed obviously, and FGF treatment recovered the characteristics of primary cells. (2) Compared with the control group, the cell proliferation in the FGF treatment group was significantly increased (P < 0.05). (3) Compared with the control group, the expression of osteogenic, adipogenic and chondrogenic related genes (RUNX2, ALP, OCN; PPARγ2, AP2, ADIPOQ; SOX9, collagen II, aggrecan) was increased significantly in the FGF treatment group (P < 0.05). The protein expressions of RUNX2,PPARγ2, SOX9 were also higher in the FGF treatment group than the control group (P < 0.05). (4) Compared with the control group, the number of extracellular calcium nodules, the number of intracellular lipid droplets, and the expression of acid acidic mucopolysaccharide were significantly increased after FGF pretreatment. To conclude, FGF pretreatment can preserve the stemness of BMSCs serially passaged in vitro.

The amount of acupuncture treatment refers to the sum of acupuncture effects during the treatment. From the factors that constitutes the amount of acupuncture treatment, including the numbers of needling, the size of acupuncture instruments, the depth of acupuncture, the manipulation, the retained time of needle, etc., and combined with teaching and clinical experience, it is believed that the amount of acupuncture treatment is closely related with clinical efficacy, besides, moderate amount of acupuncture treatment is a key factor to improve acupuncture efficacy and guarantee the safety of acupuncture. The basic principles of regular treatment amount are arrival of qi and being moderate, indicating the retained time of needle, treatment frequency and course should be based on individual condition, time, location and disease.
Acupuncture Points ; Acupuncture Therapy ; instrumentation ; methods ; Humans ; Meridians ; Needles ; Qi

Evidence has indicated that low doses of anti-tumor regimens can induce cell apoptosis in vitro, although different regimens induce apoptosis by different mechanism and pathway. In recent years, new tumor treatment strategy has been mainly focused on inducing tumor cell apoptosis. The present review discusses the advantages and disadvantages of inducing tumor cell apoptosis. The benefit of inducing apoptosis is not to cause inflammatory reaction, but as its disadvantage, it inhibits immune responses, and the phagocytosis of apopotic bodies may result in horizontal transfer of genes (including oncogenes and other oncogenic materials), which can be one of the causes of tumor relapse. This paper proposes that the tumor treatment strategy should be turn into promoting tumor cell necrosis and inducing anti-tumor immune responses.
Antineoplastic Agents ; therapeutic use ; Apoptosis ; drug effects ; Humans ; Necrosis ; chemically induced ; Neoplasms ; drug therapy ; immunology ; pathology

Objective To explore the correlative factors and management of secondary high intraocular pressure(IOP) after vitreoretinal surgery for proliferative diabetic retinopathy(PDR). Methods The data of 51 patients(54 eyes) with PDR after vitreoretinal surgery were collected.The incidence of secondary high IOP was obtained,and the correlative factors leading to high IOP were analysed.Besides,the therapeutic effects of management were observed. Results Twenty-six patients(26 eyes) experienced postoperative high IOP,with the incidence of 48.15%(26/54).It was revealed that the preoperative retinal detachment,the combined performance of crystal phacoemulsification,the practice of intraocular tamponade and the postoperative posterioruveitis were related to the occurrence of high IOP(P

Objective To analyze the clinical efficacy,adverse events and corresponding preventive measures of high dose methotrexate in the treatment of acute lymphoblastic leukemia(ALL).Methods Ninety-two patients with ALL who hospitalized in Heamotology Department of Jinshan Hospital Affiliated to Fudan University were randomly divided into observation group and control group,and each group had 46 cases.Patients in observation group were given high dose melhotrexate + vincristine + cytoxan + pirarubicin chemotherapy treatment,and in control group were given soft red enzyme + vincristine + metacortan dracin + L-asparaginase treatment.The clinical efficacy,adverse events and the required course of remission of two groups were compared.Results The total effective rate in the observation group was 84.78％,while it was 80.43％ in the control group,and there was no statistically significant difference (x2 =0.45,P ＞ 0.05).The average remission was (1.26 ± 0.28) in the observation group,while (1.31 ± 0.31) in the control group,and no significant difference was observed (t =2.13,P ＞ 0.05).The complete remission rate of initial treatment,retreatment and refractory were 75.00％ (24/32),50.00％ (4/8) and 33.33％ (2/6) respectively in observation group,while 76.67％ (23/30),50.00％ (4/8) and 25.00％ (2/8) respectively in control group,and there was no statislically significant difference between two groups (x2 =0.98,P ＞0.05).The rate of bone marrow suppression in the observation group(23.91％) was lower than that in the control group(43.48％),while the rate of liver and kidney injury(54.35％) was higher than that inthe control group(17.39％),which showed significant difference (P ＜ 0.05).The survival rate of 5 years in observation group was 67.39％,while 45.65％ in control group,and there was statistically significant difference between two groups (P ＜ 0.05).Conclusion The high dose methotrexate in the treatment of acute lymphoblastic leukemia was proved to be effective and the adverse reactions could be tolerated.

Objective To evaluate the effect of off-pump coronary endarterectomy (CE) plus off-pump coronary artery bypass grafting (off-pump CABG) on patients with chronic total occlusion (CTO) combined with diffuse distal atherosclerosis. Methods From October 2006 to August 2009,65 CTO patients with 176 angiographically confirmed vascular stenosis or occlusive lesions, 70 of which were complete occlusion, underwent off-pump CABG. During the operation, diffuse intimal thickening distal to occlusive lesion was found, and blood flow of the bridges was unfavorable.Results Therefore endarterectomy was performed, followed by CABG. The blood flow in the bridges were 2-10 ml/min versus 14-37 ml/min before versus after endarterectomy. Pulsatility index (PI) was 5.1-15.6 versus less than 5 before versus after endarterectomy. Left ventricular ejection fraction was also improved significantly [before operation: (0.47±0.12)%, after operation: (0. 52±0.15)%, t=2.17, P＜0.05]. Peri-operative myocardial infarction occurred in 2 cases, but without significant cardiac homodynamic changes. And 23 patients underwent coronary angiography to evaluate graft patency 3-18 months after operation, all of them had favorable blood flow. Conclusions It is feasible to perform off-pump CABG plus coronary endarterectomy for patients of chronic coronary total occlusion combined with diffuse distal atherosclerosis. This treatment is safe and effective.