Functional Science Lab.Of Jining Medical College has attempted to reform the management system,share the resource,improve the resource utilization,exceed the limits of science,modify the contents of experimental courses,innovate the teaching methods,pay attention to teacher training.Through longtime research and practice,we have gained the significant achievements.

ObjectiveTo develop a quantitative method for simultaneously determining multi-components in Rhei Radix et Rhizoma using one chemical reference substance.MethodsThe contents of multi-components were calculated by the UV relative correction factors (RCFs) of chrysophanol,physcion,and rhein to emodin.ResultsThe values of RCFs at 274 nm for rhein,chrysophanol,and physcion to emodin were 0.712,0.674,and 1.051.The calibration curves were linear over the ranges of 0.02-4.08,0.02-4.12,0.07- 12.92,and 0.02-3.68 μg/mL for rhein,emodin,chrysophanol,and physcion,respectively.The contents of emodin in 18 samples were determined by the extemal standard method,and the contents of the other three anthraquinone aglycones were calculated according to their RCFs.ConclusionNo significant difference is found in comparison with the classical method,indicating that the RCFs have high reliability within their linear ranges and could be used in quality control of Rhei Radix et Rhizoma.The quantitative analysis of multi-component with a single marker is especially suitable for herbal medicines containing unstable or hard to be purified components as quality control markers.

Objective To investigate the N-terminal-pro-B-type natriuretic peptide (NT-proBNP) concentration in the difference myocardial wall motion after essential hypertension. Methods 326 patients with essential hypertension and normal heart function and 265 control subjects were measured with their NT-proBNP. According to Canau's categories, the ventricle geometries of patients with high left ventricular mass index were divided into 2 groups. Results NT-proBNP was significantly higher in patients with hypertension than in controls. After grouped by different myocardial wall motion, the patients with concentric hypertrophy were highest among all the groups; the groups with eccentric hypertrophy was secondly highest; concentric remodeling and normal geometric pattern were not significantly higher than the controls. Conclusion NT-proBNP is different in patients with different myocardial wall motion before heart failure, which may play an important role in myocardial remodeling.

Objective　To evaluate the therapeutic efficiency of reversed lamellar keratoplasty for bullous keratopathy, Terrien's marginal degeneration and ocular chemical burns.Methods　Twenty eyes of patients with bullous keratopathy, twelve eyes with Terrien's marginal degeneration, ten eyes with chemical burns were recruited in this study.Diseased tissues were removed by means of dissection.Reversed lamellae corneal tissues which had been prepared by cryopreservation were thawed and rinsed. Then the prepared posterior corneal lamellae was turned over, endothelium facing upwards. The diameter of the graft should be 0.5～1.0mm larger than that of the recipient bed. The reversed donor posterior lamellae was sutured into the host keratectomy bed with 10-0 nylon sutures continuously or interruptedly.Results　Allograft reaction episode occurred only one (chemical burn) of the total 42 eyes enrolled.The grafts of the rest 41 eyes maintained clear or semitransparent for 1～5 years. Graft edema could be observed within 2～4 weeks postoperatively. Postoperative interface separation was not found. For all 20 patients with bullous keratopathy, the symptoms of pain disappeared. The visual acuity improved 2～3 lines of Snellen among eleven of twelve patients with Terrien's marginal degeneration. The visual acuity improved 2～3 lines of Snellen with four patients with chemical burns.Conclusion　The technique of reversed lamellar keratoplasty can be applied successfully for tectonic reconstruction of corneal surface such as bullous keratopathy, Terrien' s marginal degeneration and also chemical burns.The effects of therapy was satisfactory.

Objective To explore the differences of the joint function and the patient's satisfaction between two different methods to reconstruct limb length for unilateral Crowe Ⅳ developmental dysplasia of the hip (DDH).Methods The clinical controlled study were used.21 cases with unilateral DDH Crowe Ⅳ treated with total hip arthroplasty were divided into 2 groups randomly.The patients in the first group were reconstructed the limb with the compensatory length and those in the second group were did with the real length.The patients were followed up 10 years.Two independent sample t-test was used to compare these two operations.Harris scores and SF-36 scores were used in the test.Survival analysis with Kaplan-Meier method was involved in the survival of the prosthesis and then test with Log-rank test.X-ray films of different period were compared to confirm the prosthetic loosening.Results Seventeen patients were followed up 8-10 years.There were no significant differences both in Harris score and most of the SF-36 scores at the median follow-up of 10 years.The item mental healthy of SF-36 expressed significant difference.Five patients required revision.There were 10 cases suffered with polyethylene wear,6 cases with severe osteoporosis in greater trochanter,and 3 cases with osteolytic reaction.Conclusion There are no significant differences between the two surgical method in the survival rate of prosthesis and the joint function in the median follow up.The satisfaction of the control groups is lower than the trial groups both at the early stage after the operation and the median follow up.The revision rate between the two groups is similar but the reason is different.

To investigate the relationships between constitutional types of traditional Chinese medicine (TCM) and hypertension so as to provide epidemiological evidence for the theory of correlation between constitution and disease.

Objective To explore the radiosensitivity of berberine on esophageal cancer cells under hypoxia condition.Methods MTT assay and clonogenic survival assay were used to evaluate the effect of berberine on proliferation inhibition and radiosensitivity of esophageal cancer cells,respectively.Immunofluorescence was employed to examine the expression of HIF-1.The change of cell cycle distribution and cell apoptosis was assayed by flow cytometry.The expression of HIF-1 was measured by Western blot.DNA damage was detected by γ-H2AX Foci counting.Results With a clear dose and time effect,berberine inhibited cell proliferation and enhanced cell radiosensitivity(t =3.69,P＜0.05)with a sensitizing enhancement ratio(SER)of 1.42.Berberine caused a dose-dependent decrease in HIF-1 protein expression and also significantly increased the cell apoptosis in ECA-109 population(t=4.74,P＜0.05).Compared with the radiation alone group,berberine enhanced X-ray induced DNA double chain breaks(DSB).Conclusions Berberine can increase the radiosensitivity of esophageal cell line ECA-109,which may be associated with decrease of HIF-1 expression and induction of apoptosis in ECA-109 cells.

Objective To prepare recombinant human adenovirus type 3 expressing Norovirus cap-sid protein gene(Noro-orf2). Methods The cDNA for Noro-orf2 was amplifed by RT-PCR from stool of in-fantile gastroenteritis and cloned into the adenovirus shuttle vector pBSE3CMV-egfp. The vector pBSE3CMV-Nor was linearized with EeoR Ⅴ and Not Ⅰ, and transformed into E. coil BJ5183 with lined edenovirus ge-nomic DNA pLasmid pBRAdv3 by Rsr Ⅱ. The identification of recombinant adenovirus plasmid pBRAdv3E3dNor was performed by PCR, enzyme digestion and DNA sequencing. Then pBRAdv3E3dNor was digested with AsiS Ⅰ and transfeeted into Hep-2 cells with LipofectAMINETM 2000 to package recombi-nant adenovirus particles. Results Noro-orf2 was successfully inserted into the shuttle vector. The recombi-nant adenoviral plasmid pBRAdv3E3dNor was generated by homologous recombination in E. coil BJ5183 and confirmed by PCR and enzyme digestion. The recombinant adenovirus was successfully packaged and puri-fied. Norovirus eapsid protein gene expression was confirmed in Hep-2 cells by immunecytochemistry assay. Conclusion The recombinant type 3 adenovirus expressing Norovirus eapsid protein gene was successfully constructed. This study laid a foundation for developing vaccine against Norovirus.

OBJECTIVETo compare the contents and composition of volatile oil of Wenyujin (steamed root of Curcuma wenyujin) Wenezhu (steamed rhieome of Curcuma wenyujin) and Pianjianghuang (rhizome of Curcuma wenyujin).

METHODThe volatile oils of three herbal medicines obtained by steam distillation were determined according to the method described in the appendix XD in Chinese pharmacopoeia and identified by GC-MS.

RESULTThe average contents of volatile oil of Wenyujin, Wenezhu and Pianjianghuang were 0.56, 1.99 and 0.94 microL x g(-1), respectively. A total of 60 compounds were identified by GC-MS. The amounts of chemical components separated and identified in Wenyujin, Wenezhu and Pianjianghuang were 41, 41 and 40, respectively. Three medicines share a great deal of similarity, chemically and genetically. Low boiling point essential oil compositions Wenyujin and Wenezhu were similar. While the higher boiling point essential oil compositions of Wenyujin and Wenezhu were similar. And the essential oil compositions of Wenyujin and Pianjianghuang were distinct.

CONCLUSIONAlthough coming from the same origin, the composition of essential oil from different part of the plant or various process were discrepant, which lead to rather distinct clinical efficacy.

Curcuma ; chemistry ; Gas Chromatography-Mass Spectrometry ; methods ; Herbal Medicine ; Oils, Volatile ; chemistry ; Plant Extracts ; chemistry ; Plant Roots ; chemistry ; Quality Control ; Rhizome ; chemistry

OBJECTIVETo examine the feasibilities of the method, quantitative analysis of multi-components by single marker (QAMS) for determination of four alkaloids in Xianglian preparations.

METHODA HPLC method was developed to determine the contents of four alkaloids, epiberberine, coptisine, palmatine and berberine in Xianglian preparations. Berberine was selected as the internal reference substance, and 3 relative correction factors to berberine were calculated. The contents of the four alkaloids in 16 batches of samples were determined by both external standard method and QAMS. The new method was evaluated by comparing the quantitative results with of the external standard method.

RESULTNo significant differences were found between the quantitative results of four alkaloids in 16 batches of samples determined by the two methods.

CONCLUSIONQAMS is feasible for determination of four alkaloids simultaneously, and the developed method can be used for quality control of Xiang Lian preparations.

Alkaloids ; analysis ; Berberine ; analogs & derivatives ; analysis ; standards ; Berberine Alkaloids ; analysis ; Chromatography, High Pressure Liquid ; methods ; Coptis ; chemistry ; Drugs, Chinese Herbal ; analysis ; Evaluation Studies as Topic ; Quality Control ; Reference Standards