The recombinaut porcine insulin precursor(PIP)produced by Pichia pastoris in shake-flask and 501.fermenter was investigated respectively.The results indicated that 60h induction time length and 2.0%～2.5% methanol addition every day was optimum in shake- flask.The process in 50L fermenter was consisted of batch,feed-batch and induction phases.The relationship between dry cell weight(y) and culture time (t) in growth phase(batch and feed-batch phase)could be described by model y=0.6525e~(0.1907t).Glycerol and ammonia were almost used for cell growth and maintain,and no by-product was observed in batch and fed-batch phase Only 80% ammonia and 70% methanol were used by cell in induction phase.By comparison the results of shake-flask and 50L fermenter,it was concluded that the limit- ing factor in the fermentation of shake-flask and 50L fermenter was dissolved oxygen(DO)and.carbon source,respectively.When scaling the result of shake-flask to 501.fermenter,the control strategy was adapted for 50L fermenter by increasing the feed rate of methanol and the maximum PIP concentration reached 1.72 g/L.

13C metabolic flux analysis(13CMFA)have been the research hotspots of metabolic engineering internationally due to its accuracy and applicability.It is vital that the measurement of 13C labelling pattern of proteinogenic amino acids for 13C metabolic flux analysis.To acquire 13C-labelling proteinogenic amino acids,Pseudomonas denitrifican which products Vitamin B12 was firstly fed with mimimal culture medium contained 20% U-13C and 80% natural glucose,after the culture reached a steady state,then about 20 mg biomass was hydrolyzed by 1 ml of 6 mol/L hydrochloric acid for 24h at 110℃.Then amino acids was separated,concentrated,evaporated in a vacuum,and derivatized with MBDSTFA,TBDMS-derivatized amino acids can be observed by GC-MS last we get 13C labelling pattern of fifteen aminio acids through mass spectrum.The experimental methods and sample preparation offers referential value for the development of 13C metabolic flux analysis in our courtry.

Adult ; Carcinoma, Small Cell ; pathology ; Ethmoid Sinus ; pathology ; Frontal Lobe ; pathology ; Humans ; Male ; Maxillary Sinus Neoplasms ; pathology ; Paranasal Sinus Neoplasms ; pathology ; Skull Base ; pathology

Chromosomes, Human, Y ; Deleted in Azoospermia 1 Protein ; Gene Deletion ; Humans ; Infertility, Male ; genetics ; Male ; RNA-Binding Proteins ; genetics

OBJECTIVETo compare the therapeutic differences among scalp acupuncture combined with auricular point sticking, body acupuncture and western medication for treatment of vascular dementia (VD).

METHODSNinety cases were randomly divided into a combined therapy group (31 cases), a body acupuncture group (29 cases) and a western medication group (30 cases). The combined therapy group was treated with scalp acupuncture at forehead middle line, parieral middle line, temporal front line and temporal rear line as well as auricular point sticking at naogan (AT3,41), shen (CO10), shenmen (TF4), zhen (AT3), once a day; the body acupuncture group was treated with acupuncture at Baihui (GV 20), Fengchi (GB 20), Zusanli (ST 36) and so on, once a day; the western medication group was treated with oral administration of aniracetam tablets, 0.2 g per time, twice a day. Fourteen days were considered as a treatment course, and totally 3 courses were required. The mini-mental state examination (MMSE) and activities of daily living (ADL) were applied to assess the changes of cognitive behavior ability before and after treatment among three groups. Also the efficacy among three groups were compared.

RESULTSOne case dropped out in the body acupuncture group and western medication group, respectively. The total effective rate was 90.334 (28/31) in the combined therapy group, which was superior to 85.734 (24/28) in the body acupuncture group and 79.3% (23/29) in the western medication group (both P < 0.05). After the treatment, the scores of MMSE and ADL were all improved among three groups, which was the most significant in the combined therapy group (MMSE: 23.32 +/- 4.45 vs 21.23 +/- 4.13, P < 0.05; 23.32 +/- 4.45 vs 20.41 +/- 4. 01, P < 0.01; ADL: 53.18 +/- 21.55 vs 51.92 +/- 20.42, P < 0.05; 53.18 +/- 21.55 vs 49.42 +/- 19.43, P < 0.01).

CONCLUSIONThe scalp acupuncture combined with auricular point sticking could improve the clinical symptoms and cognitive behavior ability in patients with vascular dementia, which has superior total efficacy to body acupuncture and western medication aniracetam tablets.

Activities of Daily Living ; Acupuncture Points ; Acupuncture Therapy ; Aged ; Cognition ; Dementia, Vascular ; psychology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Scalp

OBJECTIVETo investigate the safety and efficacy of the transabdominal-hiatal approach of radical gastrectomy for the advanced proximal gastric cancer.

METHODSThe clinical data of 40 advanced proximal gastric cancer patients with involvement of distal esophagus admitted in Tongcheng People's Hospital from June 2003 to September 2006 were analyzed retrospectively. Patients included 27 men and 13 women with a mean age of 56.1 years (range: 37 to 76 years). The data were compared with those of 40 proximal gastric cancer patients undergoing standard thoracoabdominal approach (control group) during the same period.

RESULTSAs compared with the control group, the transabdominal-hiatal approach showed less blood loss [(181.3 ± 63.7) ml vs.(248.8 ± 79.7) ml], shorter operating time [(4.1 ± 0.6) h vs. (5.3 ± 0.7) h], shorter ICU stay [(6.5 ± 5.7) d vs. (19.4 ± 18.0) d] and shorter postoperative hospital stay [(14.0 ± 2.3) d vs.(18.7 ± 3.0) d] (all P<0.05). The complication rates of transabdominal-hiatal approach group and thoracoabdominal approach group were 7.5% (3/40) and 10.0% (4/40), and the 5-year survival rates were 51.3% and 60.0%, respectively (both P>0.05).

CONCLUSIONThe transabdominal-hiatal approach of radical gastrectomy for proximal gastric cancer is safe and effective, which may substitute the standard thoracoabdominal technique.

Adult ; Aged ; Female ; Follow-Up Studies ; Gastrectomy ; methods ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Stomach Neoplasms ; surgery ; Treatment Outcome

OBJECTIVETo investigate the correlation of male infertility with abnormality of chromosomal quantity and construction and with the deletion of DAZ gene copy in the AZFc region of Y chromosome.

METHODSIncluded in the study were 247 azoospermic and 206 severe oligozoospermic patients, as well as 210 fertile men as controls. Multi-PCR and PCR-RFLP were used to analyze the deletion of DAZ gene copies in the AZFc region of Y chromosome. Chromosomal quantity and construction were detected by G-band in the 453 patients.

RESULTSIn the azoospermic and severe oligozoospermic patients, the incidences of chromosomal abnormality were 12.6% and 8.3%; the rates of complete DAZ deletion were 7.7% and 11.2%, and the rates of DAZ1/DAZ2 deletion were 7.3% and 4.9% respectively, but no deletion was detected in the controls.

CONCLUSIONThere is a high frequency of chromosomal abnormality and DAZ gene copy deletion in patients with azoospermia and oligospermia, which suggests that chromosomal abnormality and partial and complete deletion of DAZ gene copy might be important genetic causes of Chinese male infertility.

China ; epidemiology ; Chromosome Deletion ; Chromosomes, Human, Y ; genetics ; Deleted in Azoospermia 1 Protein ; Gene Dosage ; Humans ; Infertility, Male ; epidemiology ; genetics ; Male ; Oligospermia ; epidemiology ; genetics ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; RNA-Binding Proteins ; genetics ; Sex Chromosome Aberrations

Objective To assess the feasibility,the risks and the advantages of endoscope-assisted submandibular gland resection using a retroauricular hairline incision ( RAHI) by comparing it with the conventional submandibular gland resection.Methods Twenty eight patients with benign lesions of the submandibular gland were included in the prospective clinically controlled study.Thirteen patients had endoscope-assisted resection using the RAHI approach and 15 cases had conventional transcervical approach resection.The size,location and adjacency of all lesions were evaluated by CT or MRI before surgery.The pathologic diagnoses of all cases were identified as benign diseases using fine needle aspiration biopsy.The two groups were compared for incision length,operation time,bleeding,incision cosmetic result,and complications.Results All 28 operations were successfully performed.Incision length in the endoscopic group was significantly longer than that in the trancervical group (Z =-4.516,P＜0.01),and the surgical time was longer in the endoscopic group( Z =-3.263 ,P ＜0.01) .After three months the mean subjective satisfaction score for the incision scar in the endoscopic group was significantly higher than that in the trancervical group(Z=-4.472,P＜0.01).In the endoscopic group,2 cases (15.4%) with temporary numbness of the earlobe and 1 case (7.7% ) with a temporary marginal mandibular nerve paralysis were found postoperatively.However,they recovered within 1 month.All 28 patients were disease free with a follow-up of 10 to 24 months (median of 18 months).Conclusions Endoscope-assisted submandibular gland resection via RAHI is feasible and safe for the treatment of benign submandibular gland lesions.In comparison with the transcervical approach,this method can provide better cosmetic results without significant complications.

OBJECTIVETo explore the safety and short-term efficacy of laparoscopic transhiatal proximal gastrectomy in patients with adenocarcinoma of the esophagogastric junction.

METHODSFrom Aug 2008 to May 2011, 98 patients with adenocarcinoma of the esophagogastric junction underwent laparoscopic transhiatal proximal gastrectomy. Clinical data were analyzed retrospectively including operative time, estimated bleeding, length of resection, lymph node dissection, and short-term postoperative complications.

RESULTSNinety-six patients underwent laparoscopic transhiatal proximal gastrectomy successfully and 2 were converted to open operation (one for combined splenectomy and the other combined splenectomy and resection of the tail of the pancreas). The mean operative time was (224.1±33.7) min and the mean blood loss was (69.4±26.1) ml. The mean length of esophageal resection was (4.0±0.6) cm and the resection margin was negative. The number of lymph node removed was 16.4±5.7. Pleural laceration occurred in 14 cases and spleen injury occurred in 3 case during operation. There was one anastomotic leakage. There were no postoperative mortalities, bleeding, anastomotic stenosis and wound infection. After follow-up ranging from 3 to 30 months, the value of reflux diagnostic questionnaire (RDQ) was 9.9±4.4 at 1 month and 9.3±4.3 at 3 months postoperatively. No incision metastasis was found and 5 patients died.

CONCLUSIONLaparoscopic transhiatal proximal gastrectomy is safe for patients with adenocarcinoma of the esophagogastric junction and the short-term clinical outcomes are favorable.

Adenocarcinoma ; surgery ; Adult ; Aged ; Aged, 80 and over ; Esophagogastric Junction ; pathology ; Female ; Follow-Up Studies ; Gastrectomy ; methods ; Humans ; Laparoscopy ; methods ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

Autistic traits including social reciprocal deficits, communication deficits and stereotyped behaviors, are manifested not only in patients with autism spectrum disorders (ASD) and their families, but also in general population. In recent years, there has much research related to autistic traits. This article summarizes research advance of autistic traits in ASD relations and general population.
Child Development Disorders, Pervasive ; diagnosis ; Humans ; Magnetic Resonance Imaging ; Phenotype ; Severity of Illness Index ; Surveys and Questionnaires