AIM:To simultaneously determine jatrorrhizine hydrochloride,palmatine hydrochloride,berberine hydrochloride、 evodiamine and rutaecarpine in Zuojin Pill(Rhizoma Coptidis,Fructus Evodiae).METHODS:A RP-HPLC method was established with gradient elution and an agilent C_ 18 column(250 mm?4.6 mm,5 ?m)was used.The mobile phase A was acetonitrile and the mobile phase B was water with 0.3% phosphoric acid-triethylamine.The detective wavelength was at 225 nm.RESULTS:Jatrorrhizine hydrochloride had a good linearity in the range of 0.013-0.065 ?g.The average recovery was 103.1% with RSD of 1.1%.Palmatine hydrochloride had a good linearity in the range of 0.013-0.102 ?g.The average recovery was 99.8% with RSD of 1.9%.Berberine hydrochloride had a good linearity in the range of 0.063-0.317 ?g.The average recovery was 99.8% with RSD of 1.4%.Evodiamine had a good linearity in the range of 0.001-0.007 ?g.The averoge recovery was 100.5% with RSD of 1.5%.Rutaecarpine had a good linearity in the range of 0.002-0.010 ?g.The average recovery was 98.9% with RSD of 1.9%.CONCLUSION:This method is simple,accurate and reproducible.It can be used to determine jatrorrhizine hydrochloride、 palmatine hydrochloride、 berberine hydrochloride、 evodiamine and rutaecarpine together with the same chromatogram condition.

Objective To analyze prophylactic chemotherapy outcome and clinical characteristics in patients of high-risk hydatidiform mole. Methods Twenty-three patients who were diagnosed as high-risk hydatidiform mole and undergone prophylactic chemotherapy in our hospital were retrospectively analyzed.After prophylactic chemotherapy, 11 patients didn't develop to gestational trophoblastic neoplasia (GTN),while the other 12 patients developed to GTN and needed a regimen change to combination chemotherapy.The clinical characteristics of these patients and outcome of prophylactic chemotherapy were compared between two groups. Results There was no significant difference between the two groups on patients' age,weeks of delayed menses, enlarged uterine size excessive for gestational age, and incidence of theca-lutein cysts of ovaries. However,the median levels of pre-evacuation serum β-hCG in two groups were 469 144 U/L and 768 044 U/L respectively, and median days needed for β-hCG declining to normal(≤2U/L) at the first time were 71 and 120 days respectively, which were both significantly different between two groups.Analyzed with receiver operating charactristic(ROC), the level of serum β-hCG could be a predictor for prognosis. Choosing 750 000 U/L as the cut-off value, we could expect the serum β-hCG to have a specificity of 91% and a sensitivity of 58% to predict whether prophylactic chemotherapy will be successful.Conclusions For those patients who have to receive prophylactic chemotherapy because of risk factors and unavailable hCG assessments for follow-up, it's better to use double-agent or combination chemotherapy if the level of serum β-hCG reached 750 000 U/L so as to reduce therapy duration and prevent relevant chemoresistance.

Objective To analyze the clinical and pathological features of collecting duct carcinoma (CDC) of the kidney. Methods 11 patients with CDC were analyzed,among which 6 were males and 5 were females.Their age ranged from 22 to 67 years old with a mean age of 55.4 cases were found by routine health examination,4 cases were presented with gross hematuria and 3 cases had flank or abdomen pain.The CT scan showed an unclear boundary mass in kidney,with tumor sizes from 2.1 to 8.5 cm ( mean 5.6 cm).Only medullary involvement was present on CT in 3 cases,Medullary and cortical involvement in 5 cases,Medullary and pelvic involvement in 2 cases,and all involvement in 1 case.Infiltrative lesion has expanded kidney but maintains reniform contour.Contrast-enhanced CT scan showed lesion mild to moderate enhancement compared with surrounding parenchyma. Results Radical nephrectomy was performed in 8 cases and palliative nephrectomy in 3 cases.All patients were finally diagnosed by pathology.Grossly,the tumor is often appears gray or white.In HE staining,tumor showed prominent tubular or tubulaopapillary structures with desmoplasia and inflammatory reaction.Occasionally,some tumor cells take on a hobnail appearance.Immunohistochemical examination showed UEA-1 positive in all cases,EMA positive in 9 cases,PNA positive in 8 cases,and HMW-CK positive in 7 cases.Only 2 patients showed positive CD10.7 patients died within 6 to 47 months (mean 12.5 months),2 survived with tumor free for 9 months and 8 years respectively,one lost of follow-up,and one patient with distant metastasis is receiving postoperative chemotherapy. Conclusions Collecting duct carcinoma of the kidney is a rare histological type of renal cell carcinoma,whose final diagnosis depends on histopathological examination.Rapid progression,highly malignant,poor prognosis are the characteristics of this disease.

Background Leber hereditary optic neuropathy (LHON) is mitochondrial DNA (mtDNA) disease and mainly leads to optical nerve degeneration.Its primary mechanism is synthesis disorder of DN4 protein due to variation of mtDNA 11778 locus.So to construct a vector with exogenous normal ND4 and transfect into mitochondria is a key of gene therapy for LHON.Objective This study was to investigate the in vitro transfection of adeno-associated virus (AAV)-ND4 gene into mitochondria.Methods Human renal epithelial cell lines transfected adenovirus E1A (293 cells) were regularly cultured and divided into two groups.Framework plasmids of recombinant AAV-ND4 or simple AAV2 were added to the cell medium respectively.The expression of ND4 in cells were located 12,24,36 and 48 hours after transfected by Y03 dual fluorescent quantum dots staining.The positive response for ND4 showed the green fluorescence.Results Cultured 293 cells grew well with 80％ confluence.Abundant green fluorescence particles were seen in cytoplasm in the AAV-ND4 transfected group,but only red fluorescence from mitochondrial protein was seen in the simple AAV transfected group under the fluorescence microscope.Conclusions Exogenous ND4 protein can been successfully transfected into mitochondria using the ND4 gene constructed AAV.This result provides experimental evidence for the further study on gene therapy of LHON.

Objective To investigate the mechanism and effect of the spinal cord outlet of the skull base on Chiari Ⅰ malformation with syringomyelia.Methods The cervical spinal cord stem angle (Anbc),slope angle of cervical vertebra (Ansc) of Chiari Ⅰ malformation were measured.In foramen magnum (Llf) and anterior vertebral canal level (Laf),spinal canal(Ac),spinal cord (As) and inferior hernia area (Ah) were measured.Angle,area and ratio were compared in Chiari Ⅰ malformation with syringomyelia,Chiari Ⅰ malformation without syringomyelia and normal control group.Results Ansc,Anbc-Ansc had significant differences among control group and Chiari Ⅰ malformation patients (all P＜0.001).In Llf,Laf,As had significant differences among three groups (all P＜0.05),further comparison of the two showed there were significant differences between Chiari Ⅰ malformation with syringomyelia patients and control group,Chiari Ⅰ malformation without syringomyelia patients and control group in Llf(all P＜0.05).In Llf,Laf,Ac in Chiari Ⅰ malformation with syringomyelia was smaller than control group (P＜0.05).Ah in Llf,Lafand Lh in Llf had no statistical significant difference between Chiari Ⅰ malformation with and without syringomyelia patients (all P＞0.05).In Llf,Laf,As/Ac had statistical significant difference among Chiari Ⅰ malformation with and without syringomyelia patients,control group (all P＜0.001),further comparison of the two showed As/Ac in Llf had statistical significance difference between Chiari Ⅰ malformation with syringomyelia patients and control group (P＜0.05),As/Ac in La had statistical significance difference between Chiari Ⅰ malformation with syringomyelia patients and control group,between Chiari Ⅰ malformation without syringomyelia patients and control group (all P＜0.05),Conclusion The cervical spinal cord,Ansc reducing,narrow vertebral proportion increase are important factors to promote Chiari Ⅰ malformation syringomyelia.

BACKGROUND:Microencapsulated cels are commonly used as a tool to overcome immune rejection after subarachnoid transplantation. However, the effect of microencapsulation on the secretion of human pheochromocytoma cels is unclear.
OBJECTIVE:To observe the growth and secretion of primarily microencapsulated cultured human pheochromocytoma cels in artificial cerebrospinal fluid.
METHODS: The human pheochromocytoma tissues were digested successively to isolate human pheochromocytoma cels that were then cultured in artificial cerebrospinal fluid. Primary cels were covered with alginate-polylysine-alginate microcapsules, and then the cel morphology was observed with inverted phase contrast microscope. Levels of met-enkephalin and norepinephrine in cel culture medium were detected by enzyme-labeled immunosorbent assay (ELISA). We used cel counting kit-8 colorimetric assay to obtain the growth curve of human pheochromocytoma cels in artificial cerebrospinal fluid.
RESULTS AND CONCLUSION:Microcapsulated human pheochromocytoma cels were in suspension and the process outgrowth increased slowly. Compared with non-microcapsulated cels, the proliferation rate of microcapsulated cels increased significantly. ELISA results revealed a significant increase in the levels of met-enkephalin and norepinephrine secreted from the microencapsulated cels compared to the non-microcapsule group. There was a wide variation in contents of met-enkephalin and norepinephrine from different tumors. These findings indicate that microencapsulated human pheochromocytoma cels can survive wel and have good secretion function in artificial cerebrospinal fluid, and human pheochromocytoma cels from different tumor tissues have stable secretory function.

Objective To evaluate the relationship between placental expression of Gab1 and neonatal birth weight in mothers with gestational diabetes mellitus (GDM).Methods From the singleton and full-term cesarean delivered women in Shengjing Hospital Affiliated to China Medical University between October 2014 and May 2015,30 macrosomia babies with maternal GDM were selected as GDM macrosomia group,30 cases of GDM with normal neonatal birth weight as GDM normal group,30 cases without GDM but with macrosomia as normal macrosomia group,and 30 cases without GDM and with normal neonatal birth weight as the normal control group.Gab1 protein and mRNA expression in placentas were detected using immunohistochemistry,Western blot and real-time quantitative-polymerase chain reaction.Analysis of variance,LSD,Dunnett's T3,Chi-square test and Pearson's correlation analysis were used for statistical analysis.Results (1) Gab 1 protein location and positive expression rate:Gab 1 protein expression in human placenta tissue was located in the nucleus.The positive epression rate of Gab 1 protein in the GDM macrosomia group was higher than in the GDM normal group and normal macrosomia group [93％(28/30),73％(22/30) vs 73％(22/30)] and those in the normal macrosomia group and GDM normal group were higher than in the normal control group[47％(14/30)](x2=4.320,4.320,4.444 and 4.444,all P＜0.05).(2) The expression levels of Gabl protein and mRNA:The expression level of Gab1 protein in the GDM macrosomia group was higher than in the GDM normal group and normal macrosomia group (1.43 ± 0.58 vs 1.05 ± 0.67 and 0.95± 0.59),and that in the normal macrosomia group and GDM normal group were higher than in the normal control group (0.64±0.38) (LSD test,all P＜0.05).The expression levels of Gab1 mRNA showed the same trend as the expression levels of Gab1 protein in the four groups.(3) Gab 1 protein expression level was positively associated with neonatal birth weight (r=0.320,P=0.320).Conclusions The expression of Gab1 in placenta is involved in the regulation of birth weight in GDM mothers.

This study was aimed to evaluate the feasibility of wind-warm lung-heat disease (non-severe community-acquired pneumonia, CAP) from the clinical curative effect, hospitalization days, medical expenses and other aspects. This study selected 198 wind-warm lung-heat disease (non-severe CAP) inpatients from January, 2012 to December, 2012 with the conventional therapy in the Respiratory Department, Wuhan Integrated Traditional Chinese Medicine (TCM) & Western Medicine Hospital as the control group. And another 179 wind-warm lung-heat disease (non-severe CAP) inpatients were selected from January, 2013 to December, 2013 with the TCM clinical pathway treatment in the Respiratory Department of the same hospital as the treatment group. Retrospective analysis was used to compare the clinical curative effects, hospitalization days, and medical cost differences. The results showed that TCM clinical pathway for wind-warm lung-heat disease (non-severe CAP) can shorten the hospitalization days and reduce patients’ medical costs, and ensure the clinical curative effects. It was concluded that the standardized TCM clinical pathway had positive effect on wind-warm lung-heat disease (non-severe CAP). Therefore, the formulation and implementation of TCM clinical pathway was of great significance in the treatment of wind-warm lung-heat disease (non-severe CAP).

Objective To observe the therapeutic effect of traditional Chinese medicine(TCM)granules made by the formula of Shegan Mahuang decoction for treatment of patients with asthma. Methods A prospective study was conducted,51 cases of light to moderate asthma(cold type)patients were randomly divided into treatment group (25 cases)and control group(26 cases). Conventional symptomatic treatments were of the same in two groups. The patients in control group were given salbutamol aerosol inhalation,100μg each time,4 times per day;based on the treatment of the control group,additionally the above mentioned TCM granules dissolved in water was administered orally 1 dose/day taken twice once in the morning and another once in the evening in treatment group. The therapeutic course was 12 weeks in both groups. Before and after treatment,the asthma control test(ACT)score,forced expiratory volume in 1 second(FEV1)and peak expiratory flow(PEF)were observed. The therapeutic effect,adverse reaction and compliance were evaluated. Results The number of patients with asthma completely controlled in the treatment group was significantly higher than that in the control group(16 cases vs. 9 cases,χ2=4.82,P=0.028). In the patients of two groups,FEV1,PEF and ACT scores were significantly improved after treatment,and in the treatment group,they were obviously higher than those in control group〔FEV1(L):2.67±0.46 vs. 2.32±0.47,PEF(L/min):327.6±49.4 vs. 273.1±42.8,ACT score:22.8±2.0 vs. 21.1±2.6,all P<0.05〕. After treatment,FEV1%predicted value was increased in the treatment group but decreased in control group,there was significantly statistical difference〔(0.68±0.07)% vs.(0.80±0.11)%,P<0.05〕,no significant adverse reaction was found,compliance was good. Conclusion TCM granules made by formula of Shegan Mahuang decoction can improve the clinical symptoms, pulmonary function and control level in patients with asthma,so it is an effective medicine for the control of asthma.

Objective To evaluate the diagnostic value of gastrin?17( G?17) and pepsinogen( PG) for gastric cancer. Methods A multicenter cross?sectional study of patients with continuous stomach discomfort from four centers including Changhai Hospital Affiliated to Second Military Medical University, the First Hospital Affiliated to Anhui Medical University, Qinghai Provincial People′s Hospital and the First Hospital Affiliated to Zhejiang University of Chinese Medicine from May 2014 to September 2015 was conducted. Before gastroscopy, fasting serum gatrin?17 and pepsinogen were analyzed by enzyme?linked immunosorbent assay(ELISA). The efficacy of G?17 and PG were evaluated according to endoscopic and pathological results. Results Based on the results of the pathological diagnosis, 1 122 cases were enrolled and divided into chronic atrophic gastritis group ( 548 cases ) , chronic non?atrophic gastritis group ( 370 cases), and gastric cancer group(204 cases). Serum G?17 and PGⅡ levels significantly increased(P<0?05) and PGR significantly decreased( P<0?05) in gastric cancer group compared with other groups. There was no significant difference in PGⅠlevel among three groups. The cut?off value of G?17 to diagnose gastric cancer was 7 pmol/L. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of G?17 for gastric cancer were 59?31%, 70?59%, 68?54%, 30?95% and 88?65% respectively. The cut?off value of PG Ⅰ/PG Ⅱ( PGR ) to diagnose gastric cancer was 7. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of PGR for gastric cancer were 41?18%, 83?01%, 75?40%, 35?00% and 86?39% respectively. The cut?off value of PGⅡto diagnose gastric cancer was 10 μg/L. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of PGⅡfor gastric cancer were 73?53%, 53?05%, 56?77%, 25?82% and 90?02% respectively. If G?17>7 pmol/L and PGR<7 was regarded as the cut?off value of diagnosis of gastric cancer, the sensitivity, specificity, accuracy, positive predictive value and negative predictive value were 25?00%, 91?29%, 79?23%, 38?93%and 84?56%respectively. If G?17>7 pmol/L and PGⅡ>10μg/L was regarded as the cut?off value, the sensitivity, specificity, accuracy, positive predictive value and negative predictive value were 48?04%, 79?74%, 73?98%, 34?51% and 87?35% respectively. If PGR<7 and PGⅡ>10 μg/L was regarded as the cut?off value, the sensitivity, specificity, accuracy, positive predictive value and negative predictive value were 33?82%, 84?86%, 75?58%, 33?17% and 85?23% respectively. Based on logistic regression analysis of the independent variables of high serum G?17 value(>7 pmol/L), low serum PGR value(<7) and high serum PGⅡvalue(>10 μg/L), their OR value were 2?592, 2?237 and 1?864 respectively, and high serum G?17 value showed the highest risk of gastric cancer. Conclusion High serum G?17 and PGⅡ, low PGR are indicators of gastric cancer. Combination of G?17 and PGR has the best diagnostic value for gastric cacer. Gastric cancer can be screened in large scale by combining G?17 and PGR in order to improve the early diagnostic rate of gastric cancer and reduce the mortality of gastric cancer in our country.