Objective:To explore antitumor mechanism of a recombinant vaccinia virus expressing human carcinomembryonic antigen(rV CEA).Methods:C57bl/6 mice were immunized three times with rV CEA.Six weeks later,the splenocytes from rV CEA immunized donors were adoptively tranfered in sublethally irradiated CEA + HePa bearing recipients,meanwhile the antiumor effect of the donor splencytes in virtro was detected.Results:Splenocytes from rV CEA immunized donors expressed strong antiumor activity in CEA positive tumorbearing recipients,whereas W VV immunized and nonimmune donor cell did not.The antiumor activity was abolished when immune T cells were depleted before splencytes transfer.Conclusion:It was demonstrated that antiumor acivity induced by rV CEA was essentially mediated by immune T cells which were activated by CEA specific immune response.Adoptive immunotherapy strategies may have implications for both the study of rV CEA and the development of potential clinical applications for tumor immunotherapy.

Objective To discuss the characteristics and management of pregnancy on cesarean section scar. Methods Five women with pregnancy on cesarean section scar were reviewed from Apr. 2000 to Mar. 2003. Results All of the five patients had cesarean section before and suffered from painless,irregular vaginal bleeding after 6～8 weeks of amenorrhea. The pregnancy could be misdiagnosed as invasive mole or cervical pregnancy. All had better outcome by uterine suction under ultrasound monitoring without massive bleeding or hysterectomy. Conclusions Women who had cesarean section before are in the risk of pregnancy on the old scar. Clear diagnosis should be made before any treatment. Ultrasound monitoring,local MTX injection and packing can be utilized to prevent perforation during suction. Patients should be monitored continuously after operation with serum hCG level and ultrasound. The principle prevention of pregnancy on cesarean section scar is to reduce the cesarean section rate and effective contraception.

Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Bloodletting ; Combined Modality Therapy ; Facial Paralysis ; therapy ; Female ; Humans ; Male ; Middle Aged ; Mouth Mucosa ; blood supply ; Young Adult

Objective: To study the psychological status and influencing factors in patients with nephritic syndrome(NS) before and after percutaneous renal biopsy(PRB).Methods: 235 cases of NS were examined by Symptom Check List-90(SCL-90) and STAI 24 hours before and 6 hours after PRB.Results: Before PRB,factorial anxiety and phobic anxiety were higher than norm,while after the procedure,only somatization score was still higher than norm;In men,there was a higher score than that of women after PRB,all indices were seen decreased significantly;Higher scores were seen in under-60 years old than over-60 years old.Conclusions: Before and after PRB,many factors were involved in and responsible for patients' psychological symptoms.

Objective To compare the efficacy of alfentanil and remifentanil in minimizing the propofol injection pain. Methods A total of 175 adult female patients undergoing gynecological procedures with general anesthesia were randomly divided into four groups.Patients received alfentanil 1mg(2 mL,AL group,n=43),remifentanil 0.01 mg(2 mL,REM1 group,n=43),remifentanil 0.02 mg(2 mL,REM2 group,n=45) or normal saline(2 mL,control group,n=44) 30 seconds prior to propofol administration.Visual analogue scale(VAS) was employed to evaluate the subjective feelings of pain due to propofol injection,and adverse effects were recorded. Results One patient in REM2 group and one patient in control group were excluded due to difficulty in venous catheterization.The injection pain in AL group,REM1 group and REM2 group was significantly less severe than that in control group(P

Objective To evaluate the efficacy of Qingfei-Jiedu decoction in the treatment of Acne. Methods A total of 93 female patients with acne were randomly divided into 3 groups according to random number table, 31 in each group. The observation group took the oral Qingfei-Jiedu decoction, and the Chinese medicine control group took the oral tanshinone capsule, and the western medicine control group took the oral acetic acid ring propyl alcohol progesterone. All 3 groups were treated for 4 weeks. The levels of serum dihydro (DHT), IL-6 and IL-8 were measured by ELISA. The changes of the lesions before and after treatment were observed and the adverse events were recorded during the treatment. The clinical effect was evaluated. Results After treatment, the serum DHT (56.94 ± 15.74 pg/ml vs. 74.48 ± 18.53 pg/ml, t=2.089), IL-6 (22.84 ± 11.77 pg/ml vs. 30.37 ± 14.50 pg/ml, t=2.135), IL-8 (22.64 ± 7.38 pg/ml vs. 29.54 ± 9.65 pg/ml, t=2.057) in the observation group were significantly lower than those in the Chinese medicine control group (P<0.05). After treatment, the disappearance rate of skin lesions (65.4% ± 15.0% vs. 44.7% ± 12.9%, 42.7% ± 13.6%, F=6.862) and comprehensive syndrome effective rate (57.2% ± 12.6% vs. 46.3% ± 12.8%, 44.8% ± 11.7%, F=5.322) in the observation group were significantly higher than those in the other two control groups (P<0.05). The observation group and the traditional Chinese medicine control group did not have adverse events, but the western medicine control group showed one case nausea. Conclusions The Qingfei-Jiedu decoction can reduce the level of serum DHT, IL-6 and IL-8 in female patients with acne vulgaris, and improve the skin lesion disappearance rates and comprehensive syndrome rates.

Objective To investigate the relationship between inflammatory factors,including C-reactive protein( CRP),tumor necrosis factor alpha (TNF-α),adiponectin,leptin and gestational diabetes mellitus (GDM) and their changes in puerperiunr Methods From June 2008 to May 2010,40 cases with gestational diabetes mellitus,40 cases of normal pregnancy were enrolled in this study.Fasting venous blood were obtained at early pregnancy (10 - 12 weeks),late pregnancy (36 -38 weeks) and day 3 and 42 of postpartum.Serumal adiponectin,leptin and TNF-α were measured by ELISA,and serumal CRP were measured by particle enhanced immunoturbidimetric method.The levels of those 4 cytokines and homeostasis model assessment insulin resistance (HOMA-IR) index were compared between two groups.And the changes of 4 cytokines were calculated in puerperium.The receiver operating characteristic (ROC) for TNF-α predicting GDM was designed.Results ( 1 ) The relationship between cytokines and HOMA-IR:the levels of adiponectin of (5.7 ± 1.8 ) mg/L in the GDM group were significantly lower than ( 8.1 ± 2.7 ) mg/L in control group in early pregnancy,and the level of adiponectin was negatively correlated with HOMA-IR (r=-0.333,P＜0.05).The levels of (28 ± 10) μg/L of leptin,( 10.0 ±3.4) ng/L of TNF-α and (4.7 ± 1.1 ) mg/L of CRP in GDM group were significantly higher than ( 20 ± 8 ) μ,g/L of leptin,(4.6 ±2.7) ng/L of TNF-α,(2.4 ± 1.2 ) mg/L of CRP in control group,which were positively correlated with HOMA-IR ( r =0.411,0.529,0.308,all P ＜ 0.05 ).In late pregnancy,the level of adiponectin ( 3.9 ±2.2) mg/L in the GDM group was significantly lower than ( 6.6 ± 2.7 ) mg/L in control group,and the level of adiponectin was negatively correlated with HOMA-IR ( r =- 0.344,P ＜ 0.05 ).The levels of ( 37 ±13) μg/L of leptin,( 12.7 ±2.6) ng/L of TNF-α and (6.7 ±3.6) mg/L of CRP in the GDM group were significantly higher than ( 30 ± 13 ) μg/L of leptin,(5.8 ± 2.1 ) ng/L of TNF-α,( 4.4 ± 3.1 ) mg/L of CRP in control group,which were positively correlated with HOMA-IR (r =0.414,0.487,0.285,all P ＜0.05).Multiple linear regression analysis showed that the level of TNF-α at early and late pregnancy was most correlated with HOMA-IR ( r =0.390,0.284,all P ＜ 0.05 ).( 2 ) The level of the cytokines and HOMA-IR in puerperium:at day 3 of postpartum,the level of adiponectin of (3.3 ± 1.1) mg/L in the GDM group was significantly lower than (6.2 ± 1.5 ) mg/L in control group,which was negatively correlated with HOMA-1R (r =- 0.283,P ＜ 0.05 ).The levels of ( 31 ± 13 ) μg/L of leptin,( 10.1 ± 5.7 ) ng/L of TNF-α and (35.1 ±6.5) mg/L of CRP in the GDM group were significantly higher than (21 ± 15) μg/L of leptin,(5.6 ± 3.0) ng/L of TNF-α,(30.5 ± 8.5) mg/L of CRP in control group.And leptin and TNF-α levels were positively correlated with HOMA-IR (r=0.372,0.494,all P ＜0.05).At day 42 of postpartum,the level of adiponectin in GDM group was negatively correlated with HOMA-IR ( r =- 0.299,P ＜ 0.05 ),and the levels of leptin and TNF-α were positively correlated with HOMA-IR ( r =0.401,0.442,all P ＜0.05 ).Multiple linear regression analysis showed that the level of TNF-αt at day 3 and day 42 was most correlated with HOMA-IR ( r =0.363,0.274,all P ＜ 0.05 ).( 3 ) ROC analysis of data from early pregnancy showed that the threshold for TNF-α to predict GDM was 5.45 ug/L.Conclusion TNF-ot might be the index to predict GDM and evaluate prognosis.

Objective To construct the lentivirus vector containing the hsa‐miR‐424 gene ,and identify the expression level of miR‐424 in cells .Research the influence of hsa‐miR‐424 on proliferation of cervical cancer Hela cell line .Methods Using the human genomic DNA as template to design the upper and lower primers for synthesis of miR‐424 ,and amplifying the target fragment by polymerase chain reaction (PCR) .Recover the products and conduct sequencing after connecting it into the pMD18T vector .Ampli‐fy the product by PCR template as pMD18T‐miR424 ,and insert the fragment expressing pMD18T‐miR424 into the vector of pLen‐tis‐CMV‐GFP‐MCS‐PGK‐PURO after enzyme cutting to construct the pLentis‐CMV‐GFP‐miR424‐PGK‐PURO .Package the com‐pound with pMD2 .G and pSPAX2 in 293T cell to produce the lentivirus ,and using the supernatant containing lentivirus to infect the Hela cell line .Results The sequencing result proved the sequence of miR‐424 in plasmid vector was correct ,which proved the construction of lentivirus was successful and the target lentivirus was obtained .The expression of miR‐424 almost rise 60 times af‐ter infected the cervical cancer Hela cell by the carrier .The result of M TT method suggested :the cervical cancer Hela cell lines have slowed proliferation with infection miR‐424 lentivirus .Conclusion The miR‐424 lentivirus vector was constructed successfully and the high efficacy expression miR‐424 cell line was established and stable .The cervical cancer Hela cell were infected with the super‐natant containing lentivirus ,inhibited the proliferation of Hela cell successfully ,and laid a good foundation for subsequent research .

12E7 Antigen ; Adnexa Uteri ; surgery ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD ; metabolism ; Cell Adhesion Molecules ; metabolism ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Granulosa Cell Tumor ; drug therapy ; metabolism ; pathology ; surgery ; Humans ; Hysterectomy ; methods ; Inhibins ; metabolism ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Ovarian Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; Survival Rate ; Vimentin ; metabolism

The objectives of this study were to investigate the effects of the CRISPR/Cas9 system mediated by the HPV pseudotype virus on SiHa cytobiology behavior by cutting the HPV16 E6 gene selectively and to explore the role of this system in the treatment of cervical cancer.After designing specific gRNA sequences targeting HPV 16 E6,generating hCas9-EGFP and E6-gRNA-RFP plasmids,and preparing the pseudovirus of HPV16 carrying E6-gRNA and Cas9 plasmids,we determined the titer of the pseudotype virus using the TCID50 method.We obtained the pseudotype virus of HPV16 carrying E6-gRNA and Cas9 plasmids to transfect cervical cancer SiHa cells.Experimental subjects were divided into control group,empty virus group,E6-gRNA transfected group,Cas9 transfected group and Cas9+E6-gRNA transfected group.The molecular size of the cutting sequence was detected using the T7E1 enzyme digestion method and agarose gel electrophoresis,and the cleavage function of CRISPR/Cas9 on the E6 gene was determined at the same time.RT-PCR and Western blotting were performed to detect the mRNA and protein expression levels of E6 in all the groups;the Transwell cell migration assay was performed to detect the cell migration ability and metastasis in all groups.Heterotopic transplantation tumors were incorporated into mice and were used to investigate the effects of the CRISPR/Cas9 system mediated by the HPV pseudovirus on the tumorigenic ability of SiHa cells by selectively cutting HPV16 E6.The HPV16 pseudotype virus carrying E6-gRNA and Cas9 plasmids could successfully infect SiHa cells,and there were two cutting zones in the Cas9+E6-gRNA transfected group.However,the empty virus group,E6-gRNA transfected group and Cas9 transfected group had no corresponding zone.Compared with those in the control group,the empty virus group,E6-gRNA transfected group and Cas9 transfected group,the mRNA and protein expression levels of E6 in SiHa cells were downregulated in the Cas9+E6-gRNA transfected group (P＜0.01).In addition,the proliferation and migration abilities of SiHa cells were significantly inhibited (P＜0.01).There were no significant differences among the other groups.In contrast to the control group,the HPV pseudotype virus carrying E6-gRNA and Cas9 plasmids could significantly delay the growth of tumor cells of the ectopic tumor transplantation model (P＜0.01).The CRISPR/Cas9 system mediated by the HPV pseudotype virus to knockout E6 gene expression exhibited a clear inhibitory effect on the biological function of SiHa cells,which indicated that knocking out the E6 gene using the CRISPR/Cas9 system mediated by the HPV pseudotype virus had a potential effect of eliminating HPV infection and inhibiting the growth of HPV-related tumors.Taken together,these findings provide insight into a new treatment strategy for the prevention and treatment of hr-HPV infected disease,particularly in HPV-related tumors.