In this article,the authors reported a modified method of determining Oxygen consump- tion for the mouse hypoxia experiment in order that the students could observe the dynamic change of oxygen consumption rate(OCR)and respiratory rate(RR)in the whole process of acute progressive hypoxic hypoxia on mice.The results showed that to reach the maximal OCR and RR,especially the latter,required more time as the survival time was prolonged. In a closed set of the same size,the heavier the mouse weighed,the shorter it survived,but there was an individual difference.

Based on the work we have previously done, the experiments presented here indicate that ?aS1, a mutant of the P19 line of routine embryonal carcinoma (EC) cells, can be induced to differentiate by retinoic acid (RA), but is not sensitive to DMSO. Also there is a dynamic equilibrium achieved by a balance between the processes of ?aS1 cell proliferation and differentiation when exposed to certain RA concentrations. The ?aS1 cell has a lower ability for forming gap junctions as compared with the other mutants, showing that EC cell mutants could be defective in the development of a certain gap junction type. In the RA-treated mixed aggregates of P19 and RAJH6, only P19 cells differentiated continuously but the surviving fraction of RAJH6 cells in the presence of 6-TG kept unchanging. The result suggests that the undifferentiated and differentiating EC cells can be metabolically coupled by gap junctions. Our work may further the understanding of cell differentiation, congenital abnormality and tumor reversion.

Objective: To construct the packaging plasmid pSNAV-hEndostatin-CMV-EGFP of adeno-associated virus vector (AAV) encoding human Endostatin (hEndostatin) cDNA. Methods: The hEndostatin eDNA obtained from plasmid pCD-sEndostatin was subcloned into the packaging plasmid pSNAV of AAV by molecular clone ways. The recombinant plasmid pSNAV-hEndostatin-CMV-EGFP was identified by PCR analysis, restriction enzymes analysis and sequencing analysis.Results: The recombinant pSNAV-hEndostatin-CMV-EGFP was correctly constructed and confirmed by PCR analysis, restriction enzymes analysis and sequencing analysis. Conclusion: The constructed AAV-hEndostatin packaging plasmid pSNAV-hEndostatin-CMV-EGFP could be the packaging plasmid of rAAV-hEndostatin-EGFP.

Objective To study the expression of pericyte and the association between pericyte and angiogenesis in the endometriosis. Methods the endometriosis group is consisted of 20 ectopic endometrium, 20 eutopic endometrium dur-ing proliferation phase, 20 eutopic endometrium during secretory phase of patient with endometriosis. The control group (non-endometriosis ) include 20 normal endometrium during proliferation phase and 20 normal endometrium during prolifer-ation phase of women without entometriosis. The histological morphology parameters such as mean density of microvessel (MVD), pericytes number(PN)as well as the average ratio of pericyte to endothelial cells in microvessels were measured by immunohistochemical and morphological assesses. Results The expression of MVD was lower in the normal endometrium and eutopic endometrium compared with it in ectopic endometrium tissue. The difference of the MVD among the different groups was statistical significant (P＜0.05). The expression of MVD in secretory phase endometrium was higher than it in proliferative phase endometrium (P＜0.05), and it shows no significant difference between eutopic endometrium and normal endometrium. The expression of PN and PN/MVD in ectopic and eutopic endometrium in EMs group were lower than those in control group. However, we also found that the difference of PN and PN/MVD was not statistically significant. Conclusion Pericytes play an important role in angiogenesis of the endometriosis ,and pericyte coverage contribute to maturing of func-tional vasculature of endometriosis.

Objective To investigate the relationship between the amplification of human telomerase reverse tran-scriptase (hTERT) gene and high-risk human papillomavirus (HR-HPV) infections in cervical intraepithelial neoplasia (CIN) and cervical carcinoma. Methods The cervical epithelial cells were collected from 34 samples of normal cervical epithelium, 31 samples of CIN (gradeⅠ), 33 samples of CIN (gradeⅡ), 34 samples of CIN (gradeⅢ) and 20 samples of cer-vical carcinoma. HPV DNA was detected by polymerase chain reaction-reverse dot blot hybridization (PCR-RDB) and the amplification of hTERT gene was detected by fluorescence in situ hybridization (FISH). Results Twenty subtypes of HR-HPV were detected including HPV16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 67, 68, 69, 73 and 82. The inci-dence of HR-HPV infection was higher in CINⅡgroup (72.73%), CINⅢgroup (85.29%) and cervical carcinoma group (90.00%) than that of normal cervical epithelium group (20.59%). There was no significant difference in the positive rate of HR-HPV DNA between CINⅠ group (54.84%) and normal cervical epithelium group (P < 0.005). The positive rate of hTERT gene amplification was higher in cervical carcinoma group (80.00%) than that of normal cervical epithelium group (0). There were no significant differences in the positive rates of hTERT gene amplification between CINⅠgroup ( 3.22%), CIN Ⅱ group (18.18%), cervical carcinoma group and CIN Ⅲ group (41.18%). There was positive correlation between hTERT gene amplification and HR-HPV infection (r=0.238, P<0.05). Conclusion The incidence of HR-HPV infection was positively correlated with hTERT gene amplification in cervical lesions. HR-HPV infection may be an early event of ab-normal amplification of hTERT gene. The detection of HPV-DNA and hTERT gene can be used in the clinical diagnosis of early cervical lesions.

Objective:To investigate the effect of human interferon-α-2b(IFN-α-2b) in the treatment of endometriosis by detecting the levels of CA125 and endometrial antibody(EMAb). Methods: Forty-five cases with endometriosis were divided into three groups. Fifteen cases in test group (operation+IFN-α-2b), 20 cases in control group A (operation+Diphereline), 10 cases in control group B(only operation). The blood serum level of CA125 was detected by microparticle enzyme immunochemiluminescent at pre-operation, the first month, second month and third month of the post-operation; and the EMAb level was measured by enzyme linked immunosorbent assay (ELISA). The follow-up of patients was carried out after operation to evaluate the clinical appearances and record the adverse effects of the human IFN-α-2b. Results: Compared with pre-operation, the levels of CA125 and EMAb were obviously degraded in the first to third months after surgery in test group and control group A(P < 0.05). There were no significant difference in the levels of CA125 and EMAb were at pre-operation,the first month,second month and third month of the post-operation between the test group and control group A(P > 0.05). There were no significant difference in the levels of CA125 and EMAb at pre-operation between the test group and control group B(P > 0.05). However, the level of EMAb was lower the first month after operation in the test group than that of control group B. In the follow-up, the adverse effects of the human interferon-α-2b were observed including low-grade fever, articular muscle soreness and gastrointestinal tract complaints. These appearances completely disappeared after drug withdrawal. It had less impact on the menstruation. Conclusion: The levels of CA125 and EMAb were similar in treatment of endometriosis with human IFN-α-2b and with Diphereline. Both of the treatments are better than the pure surgery. It has less impact on the menstruation and can avoid menostasia.

The ketoreductase (KR) domain in the first extending module of the polyketide synthase (PKS) catalyzes the reductions of both an α-keto group and a β-keto group in the biosynthesis of bacillaene, suggesting the intrinsic substrate promiscuity. In order to further investigate the substrate specificity, the KR domain (BacKR1) was heterologously overexpressed in Escherichia coli. In vitro enzymatic analysis showed that only one of the four diastereomers was formed in the reduction of the racemic (±)-2-methyl-3-oxopentanoyl-N-acetylcysteamine thioester catalyzed by BacKR1. In addition, BacKR1 was revealed to catalyze the reductions of cyclohexanone and p-chloroacetophenone, indicating the potential of KR domians of PKSs as biocatalysts.
Bacterial Proteins ; genetics ; metabolism ; Catalysis ; Cyclohexanones ; metabolism ; Escherichia coli ; enzymology ; Polyketide Synthases ; genetics ; metabolism ; Protein Structure, Tertiary ; Substrate Specificity ; omega-Chloroacetophenone ; metabolism

Objective To evaluate the efficacy and safety of CT-guided 125I radioactive seed implantation combined with gemcitabine and Gio (gemcitabine and S-1, GS scheme) chemotherapy in treating advanced pancreatic carcinoma. Methods Sixty-eight patients with inoperable advanced pancreatic carcinoma were randomly divided into two groups. Patients in group A(n=38) were treated with CT-guided 125I radioactive seed implantation combined with GS chemotherapy scheme, while patients in group B (n=30) received GS chemotherapy scheme only. The short-term effect, the median progression-free survival time, the median survival time and adverse reactions of the two groups were determined , and the results were compared between the two groups. Results The objective response rate (ORR), disease control rate (DCR) and clinical benefit rate (CBR) of the group A were 57.9%, 73.7%and 84.2%respectively, while those of group B were 26.7%, 46.7% and 60.0% respectively. The differences between the two groups were statistically significant (P<0.05). In group A the median progression-free survival time and the median survival time were 8.00 months and 11.84 months respectively, which were strikingly higher than those in group B (5.63 months and 10.40 months respectively), the differences between the two groups were statistically significantly (P<0.05). No significant differences in gastrointestinal reactions, blood toxicity, liver toxicity and other adverse reactions existed between the two groups (P>0.05). Conclusion For advanced pancreatic carcinoma, CT-guided 125I radioactive seed implantation combined with GS program is a safe and effective treatment.

Objective To investigate the value of CT-guided 125I particles implantation combined with gemcitabine plus S-1 (GS) regimen in the treatment of locally advanced pancreatic cancer.Methods 42 patients with unresectable local advanced pancreatic cancer were given with CT-guided 125I seed implantation.3-4 cycles of GS regimen was given based on the tolerance of patient s body within 3-7 d after implantation of particles.Review of blood,CA199,chest X-ray,CT scan + enhanced or MRI were performed at 2nd,4th,6th,12th month after surgery.Results 2nd,4th,6th month after surgery,tumor lesions were significantly reduced,ORRs were 59.5 ％ (25/42),66.7 ％ (28/42) and 73.8 ％ (31/42),respectively,DCRs were 83.3 ％ (35/42),78.6 ％ (33/42) and 76.2 ％ (32/42),respectively.No serious adverse reactions were observed,patient could tolerate these reactions.The 6th,12th,24th month survival rates were 100 ％ (42/42),47.6 ％ (20/42) and 11.9 ％ (5/42),respectively,mPFS was 8.27 months and mOS was 12.00 months.Conclusion CT-guided 125I particles implantation combined with GS regimen is convenient,safe,high efficacy in the treatment of locally advanced pancreatic cancer.

Objective To analyze the sensitized factors in urinemia patients who waiting for renal transplantation.Methods 2429 patients with urinemia from April 2002 to December 2008 were subjected to the detection of panel reactive antibody, and classified into 5 groups according to their clinical data:(A) no history disease group (n = 1097) who never experienced transfusion, pregnancy and transplantation; (B) Transfusion group (n = 361) who received transfusion more than 200 ml; (C) Pregnancy group (n = 481) who experienced pregnancy; (D) Transfusion+ pregnancy group (n= 294) who experienced both pregnancy and transfusion; (E) Re-transplantation group (n = 196) who experienced failed transplantation before, and waited for the second renal transplantation.Results All the males in group A were negative for PRA, and females were weakly positive for HLA Ⅱ antibody.The incidence of PRA production in group B was 15.24 % (55/361).Thirty-nine patients were positive for PRA in group C with the incidence being 8.11 % (39/481).The PRA positive rate in groups D and E was 30.61 % (90/294) and 70.92 % (139/196) respectively.PRA intensity was more than 60 % in 72 patients in group E.Conclusion Transfusion and pregnancy caused lower incidence of PRA positive rate.The incidence was much higher in transfusion + pregnancy patients than that in patients with transfusion or pregnancy alone.Graft caused the higher incidence of PRA than by transfusion and pregnancy.