Objective To establish a cell line and a convenient and well-duplicated minimal residual leukemia (MRL) model. Methods We chose the international standard domestic inbred DBA/2 mouse to inoculate the different numbers of L1210 leukemic cells to investigate the relationship between inoculating cell number and mouse survival time. On the third day after 1×106 leukemic cells per mouse were inoculated, different doses of CTX were used for the chemotherapy. Results The inoculating number of L1210 leukemic cells increased while the mouse survival time decreased;The survival time increased following the increase of the doses of the CTX;The survival time of the inoculated mouse treated with CTX at 125 mg/kg was equal to that of the mouse inoculated with 500 L1210 leukemic cells without treatment. Conclusion The observation results are that the dose of 125 mg/kg for creating the model of MRL mouse.

Objective To study the clinical characteristics, diagnosis, and treatment of patients with persistent low levels of β-hCG. Methods To analyze the clinical data of 6 patients with persistent low level of β-hCG retrospectively and review the relevant literature. Results The periods of low levels of β-hCG in the 6 patients were 10-53 months. The range of β-hCG was 3～637 U/L. The last pregnancy were hydatidiform mole in 5 cases and abortion in 1 case. After hydatidiform mole 5 cases were received multi-chemotherapy. Normal level of β-hCG maintained 1-12 months after chemotherapy, then β-hCG titer has gone up again, One patient developed lung metastases after 30 months and then cured by surgery and chemotherapy. There were no clinical evidence of malignancy in the other 4 cases, and 1 case received no therapy after abortion during course of therapy and following up. Conclusion Persistent low level of hCG was a special kind of disease with poor response to chemotherapy, but the majority were with stable status. Therapy should be restrained if there was no evidence of malignancy.

Objective To observe the curative effect of complex melanteritum pill in treating anemia of hypothyroidism.Methods A total of 50 patients were randomly recrutied into a control group(20 patients)and a treatment group(30 patients).The treatment group was treated by Levothyroxine Sodium Tablets,50 μg qd,and complex melanteritum pill,nine pills/time,three times daily;while the control group was treated by levothyroxine sodium tablets,50 μg qd.Both groups were treated for 8 weeks,without blood transfusion,adding iron and folic acid and vitamin B12,etc;Results The basic cure rate and the total effective rate in the treatment group was 50.0%and 80.0%respectively;the basic cure rate and the total effective rate in the control group was 30.0%.and 50.0%respectively.There was a significant difference between the two groups by comparison(P<0.05).Conclusion Complex melanteritum pill was effective in treating hypothyroidism with few side effects.

Objective To explore the clinical effect of flupentixol and melitracen(deanxit)combined with trimebutine maleate(TMB)in the treatment of patients with functional gastrointestinal disorders(FGID).Methods 135 patients with FGID were selected,and were divided into two groups accorded to the random number method,with 68 cases in the observation group and 67 cases in the control group.The control group was treated with oryzanol and vitamin B6 ,while the observation group was treated with deanxit combine with TMB on the basis of the control group. Main symptoms of postprandial discomfort,early satiety,abdominal pain and burning sensation on the abdomen in the two groups before and after treatment were monitored,adverse reactions of two groups were recorded,and the total efficacy was evaluated after the end of treatment.Results The difference of postprandial discomfort,early satiety, abdominal pain and upper abdominal burning sensation integration between the two groups had no statistically significant before treatment(all P >0.05).After treatment,the postprandial discomfort,early satiety,the abdominal pain and epigastric burning sensation were (4.1 ±1.6)points,(3.4 ±1.1)points,(4.2 ±1.2)points and(4.4 ± 0.7)points in the observation group,which were better than (5.9 ±2.4)points,(5.3 ±1.6 )points,(6.0 ± 1.2)points and(6.1 ±0.5)points in the control group(t =3.057,4.791,5.196,9.681,all P <0.05).The total efficacy was 94.1%(64 /68)in the observation group,which was better than 77.6%(52 /67)in the control group(χ2 =7.603,P <0.05).The difference of adverse reaction rates between the two groups had no statistically significant(P >0.05).Conclusion Deanxit combine with TMB can significantly improve symptoms in patients with FGID,and has a significant effect,thus it is a safe and effective therapy.

Objective To investigate the effect of a new cloned full length gene, C2 gene, which encodes human eukaryotic translation initiation factor, on tumorigenesis of human gastric cancer cells in vivo and in vitro. Methods A constructed eukaryotic vector carrying the full length of C2 gene was amplified, purified, and transfected into a gastric cell line SGC7901 cell. The expression of C2 protein was examined with fluorescence activated cell sorting (FACS) and Western Blot. The effect of C2 gene on tumorigenesis of human gastric cancer cells was investigated in vitro and in vivo with methods of clone formation in plate and oncogenesis in nude mouse. Results FACS and Western Blot showed that C2 protein was expressed highly and stably in transfected cells. The average clone formation rate of C2 gene transducted cells is 43.8%, the rate which is lower than that of vector transducted cells (76.9%). In vivo, the time of tumorigenesis of C2 gene transducted cells in nude mouse is prolonged from 1 week to 2 weeks, and the volume of tumor node is smaller than that of vector transducted cells, that is, 1.20 and 5.58 cm 3 respectively ( P

Objective　To investigate the effect of a new cloned full length gene, C2 gene which encodes a translation initiation factor, on cell cycle and the relationship between C2gene and apoptosis. Methods　Construct a eukaryotic vector carrying the full length of C2 gene, then transfected it into a gastric cell line-SGC7901 cells and gained cells expressing C2 protein transiently or stably. Test the expression of C2 protein and the change of cell cycle of the transfected cells using the flurescence activatied cell sorting(FACS) and the changes of their ultra structure electromicroscopically. Results　The C2 gene eukaryotic expression vector was successfully constructed. FACS showed the C2 protein expression ratio of transfected cells was 32.3%, while the empty control group was 0.9%. The test of cell cycle showed that there was appeared apoptosis peak in transiently or stably transfected cells. The apoptosis cells can be seen electromicroscopically among the C2 transfected cells. Conclusions　C2 gene transfection can lead SGC7901 cells to apoptosis, but the mechanism has to be further study.

The gene encoding thermostable lactate dehydrogenase (Tm-LDH) was cloned into the plasmid pHsh from Thermotoga maritima, and expressed in Escherichia coli JM 109. The recombinant protein was purified to homogeneity by a simple step, heat treatment. The recombinant enzyme had a molecular mass of 33 kDa. The optimal temperature and pH of Tm-LDH were observed 95 degrees C and 7.0. The purified enzyme had a half-life of 2 h at 90 degrees C, and exhibited better stability over a pH range from 5.5 to 8.0. The K(m) and V(max) values were 1.7 mmol/L, 3.8 x 10(4) U/mg of protein for pyruvate, and 7.2 mmol/L and 1.1 x 10(5) U/mg for NADH, respectively. The expression of Tm-LDH in T7 system could not obtain high efficiency, but it has been soluble over-expression in pHsh system and reached 340 mg/L. The superior stability and productivity of Tm-LDH will lay the foundation of its industrial-scale fermentation and application in the NAD regeneration.
Cloning, Molecular ; Enzyme Stability ; Escherichia coli ; metabolism ; L-Lactate Dehydrogenase ; biosynthesis ; Molecular Weight ; Recombinant Proteins ; biosynthesis ; Temperature ; Thermotoga maritima ; enzymology

Objective:To observe the effect of combining Chinese medicine and tuina along the meridians on motor function and activities of daily living (ADL) in patient with post-stroke upper limb spasticity. Methods:A total of 220 patients with post-stroke upper limb spasticity were randomly allocated into a treatment group (n=110) and a control group (n=110). Patients in the treatment group received tuina along the meridians combined with spasticity-alleviating and collateral-unblocking Chinese medicine, whereas patients in the control group received routine rehabilitation therapy. Patients in both groups were treated for 3 weeks. Then the patients’ motor function, ADL and muscle tone were evaluated before and after treatment using the Fugl-Meyer assessment scale (FMA), modified Barthel index (MBI) and modified Ashworth scale (MAS). Results:After treatment, the FMA scores, MBI scores, and muscle (shoulder intortor, elbow flexors and wrist flexors) tones were significantly improved (P<0.05), but the improvement was more significant in the treatment group than that in the control group (P<0.05). Conclusion:Tuina along the meridians combined with spasticity-alleviating and collateral-unblocking Chinese medicine can substantially alleviate muscle tone on the affected side and remarkably improve the patients’ motor function and ADL.

Objective To study the expression of insulin-like growth factor-Ⅱ receptor(IGF-Ⅱ R) and its relationship with the prognosis of human epidermalgrowth factor receptor 2 (HER2) positive breast cancer.Methods The expression of IGF-Ⅱ R in 104 specimens of HER2 positive breast cancer was evaluated by immunohistochemistry.The expression was rated on 2 scales:negative and positive.The age,pathological type,tumor size,lymph node involvement,status of estrogen receptor(ER),and progesterone receptor(PR) of all the cases were reviewed.The relationship between the expression of IGF-Ⅱ R and the above pathological parameters were analyzed by statistical methods.The relationship between the expression of IGF-Ⅱ R and the disease-free survival and overall survival was studied.Results The positive expression rate of IGF-Ⅱ R was 43.3％ (45/104).The positive rate of IGF-Ⅱ R was significantly higher in the lymph node positive group than in the lymph node negative group (61.5 ％ vs 25.0％,P =0.000).No significant correlation was observed between the expression of IGF-Ⅱ R and age,pathological type,tumor size,status of ER,or PR.The disease-free survival and overall survival was significantly lower in IGF-Ⅱ R positive group than in IGF-Ⅱ R negative group.(The 5-year disease-free survival:62.6％ vs 84.7％,P =0.022; The 5-year overall survival:71.5％ vs 89.6％,P =0.024).Cox regressive analysis showed that the lymph node status was an independent risk factor of disease-free survival and overall survival,while the IGF-Ⅱ R was not.Conclusion The positive expression of IGF-Ⅱ R in HER2 positive breast cancer indicates poor prognosis.The expression of IGF-Ⅱ R may be regulated by insulin-like growth factor-Ⅱ (IGF-Ⅱ).

Objective To compare the efficacy and safety of icotinib therapy alone versus icotinib combined with thoracic radiotherapy for the treatment of advanced non-small cell lung cancer (NSCLC) patients with an activating epidermal growth factor receptor (EGFR) gene mutation.Methods A total of 83 patients with advanced NSCLC harboring an activating EGFR gene mutation was enrolled in this study.All the patients were randomly divided into 2 groups.Patients in group A (n =41) received thoracic radiotherapy (prescribed at 60-66 Gy) combined with icotinib (three times per day,125 mg once).Patients in group B (n =42) were given icotinib therapy alone (three times per day,125 mg once).Treatment was continued until disease progression or unacceptable toxicity or death.The primary end points were median progression-free survival (mPFS) and 12 month-PFS rate.The secondary end points included objective response rate (ORR),disease control rate (DCR) and adverse events.Results With a median follow-up of 18.2 months,mPFS was 15.2 months (95％ CI:12.2-17.4) in group A and 13.2 months (95％ CI:10.8-14.9) in group B (x2 =4.29,P=0.036).PFS rates of 12 months for group A and group B were 70.3％ and 61.2％,respectively.The ORR were 78.0％ vs.57.1％ (x2 =5.16,P =0.028),and the DCR were 95.1％ vs.92.9％ (P＞0.05) in groups A and group B,respectively.No grade 3-4 adverse events was observed in both groups except the rashes (4 cases in each group).Besides,10 patients had grade 1-2 radiation-related pneumonitis and 15 patients suffered grade 1-2 radiation-related oesophagitis in group A.Conclusions In advanced NSCLC patients with an activating EGFR gene mutation,the combination of thoracic radiotherapy and icotinib had achieved an improvement on ORR and PFS with good tolerance.Clinical trial registration Chinese clinical trial registry,ChiCTRINR-16010262.