Objective:To establish a method for the determination of related substances in erythromycin ophthalmic ointment. Methods:The extraction method was established and optimized. An HPLC gradient elution method was used for the determination of related substances in erythromycin ophthalmic ointment. Results: One step extraction with PBS (pH 7.0) - methanol(1: 1) had promising effect. After the method validation, it was proved that the method could be used to determine the related substances in eryth-romycin ophthalmic ointment. Conclusion:The method established in the paper provides a better analytical extraction and determina-tion method for the quality control of erythromycin ophthalmic ointment.

Objective: To investigate the relationship between transportation of apoptosis-inducing factor (AIF) from mitochondria to nucleus and DNA damage at different time points during apoptosis of SMMC-7721 cells induced by hydroxycamptothecin (HCPT). Methods: Cells were harvested at different time points after HCPT treatment. The mitochondrium, cytosol, and nucleus were isolated with mitochondria isolation kit. These fractions were used for western blot. Transportation of AIF from mitochondrium to the nucleus at different time points was observed by confocal microscopy. At the same time, DNA damage was detected by TUNEL assay. Results: Western blot showed that transportation of AIF from mitochondrium to nucleus was not observed after treatment with HCPT for 1 h. However, AIF immunoreactivity was significantly increased in the nuclear fraction at 6, 12 and 24 h after HCPT treatment. Confocal microscopy demonstrated that AIF was released from mitochondrium at 6 h after HCPT treatment. The massive transportation of AIF to the nucleus occurred at 12 and 24 h after HCPT treatment. At the same time, DNA damage was observed by TUNEL assay. Conclusion: AIF release is one of the early biochemical changes of apoptosis of SMMC-7721 cells after HCPT treatment. The DNA damage occurrs in parallel with AIF translation to nucleus.

Objective To examine mutations in the WT1 and PLCE1 gene in three Chinese families with autosomal recessive steroid-resistant nephrotic syndrome (SRNS) once mutations in NPHS2 had been excluded. Methods Peripheral blood samples were collected for genetic analysis from three probands of three Chinese families and their parents, and two probands' siblings, and 50 adult volunteers with normal urinalysis. Genomic DNA was isolated from peripheral blood leucocytes. Ten exons and exon-intron boundaries of WT1, and 31 exons and exon-intron boundaries of PLCE1 were amplified by polymerase chain reaction (PCR). Mutational analysis was performed by DNA sequencing directly and RFLP (restriction fragment length polymorphism) and/or PCR. Results No mutation in both WT1 and PLCE1 was identified in three probands from three Chinese families with autosomal recessive SRNS. However, three variants of WT1, 126C>T, ⅣS5-64A>G and 903A>G, and 13 variants of PLCE1, -134A>G, 810T>C, 960G>A, ⅣS11-28C>G, ⅣS15+26A>C, 4724G>C, ⅣS20+40C>T, ⅣS21+64G>A, ⅣS22-26T> A, 5320C>T, 5780A>G, ⅣS27+24A>G and ⅣS31 +48_49insT, were detected in three probands and some controls, indicating that all these variants were gene polymorphisms. WT1 polymorphism ⅣS5-64A>G, and PLCE1 polymorphism ⅣS22-26T>A were novel. Conclusion All the encoding exons and exon-intron boundaries of both WT1 and PLCE1 in three probands are examined, and no causative mutations in WT1 and PLCE1 axe found, suggesting that mutation in WT1 and PLCE1 genes is not a major cause of the Chinese families with autosomal recessive SRNS.

Objective The color of the syphilis quality control material adopted by most detection institutes was the same with the detected serum sample and they were all colorless,transparent or light yellow.There were cases of wrong adding,missing adding or insufficient adding due to the color of quality control materials which was hard to distinguish with naked eyes.To avoid this phenomenon,a new method was established for the distinction of quality control materials.Methods A new method of syphilis quality control materials that had been improved three concentrations control materials:0.125,0.250 and 0.500 NCU/mL.The syphilis diagnostic kit that was created by Shanghai Kehua and Xiamen Yingke was adopted to conduct detection and compare results.Results The difference between stained quality control material and unstained quality control materials had no statistical significance (P＞0.05).Two different reagents were used to detect quality control materials of different concentration for 20 times and the CV were 11.7 ％-13.4％ and 9.3 ％-12.9 ％ respectively.Two different reagents were used to detect quality control materials of different concentration for 30 days and the CV range were 10.1 ％-13.4 ％ and 8.08 ％-12.8 ％.Conclusion Citric yellow staining does not influence the properties of syphilis control materials and it can be used stably for a long time.It is suitable for clinical lab application and promotion.

T were also detected in the controls.Conclusion Nephrotic syndrome may be associated with NPHS2 identified in children.

OBJECTIVETo study the effect of hydroxycamptothecin (HCPT) on apoptosis-inducing factor (AIF) expression and AIF translocation from mitochondria to the nucleus in human hepatocellular cancer cell SMMC-7721 during apoptosis.

METHODSAfter treatment with 80 mg/ml of HCPT, the cancer cells were stained with A0/EB to monitor their apoptosis. Their mitochondria was examined with electronmicroscopy and the AIF expression of the cells was tested by RT-PCR and Western blot. The translocation of AIF from mitochondria to the nucleus during apoptosis was analyzed by confocal microscopy.

RESULTSSMMC-7721 cells treated with HCPT showed chromatin condensation, nuclear fragmentation and mitochondria swelling. The mRNA and protein expression of AIF in treated and untreated SMMC-7721 cells were not significantly different. However, cells treated with 80 mg/ml HCPT for 6 h or 12 h showed massive translocation of AIF into the nuclei.

CONCLUSIONThese results show the important role the mitochondrial pathway of apoptosis plays in HCPT-induced tumor cell death, at least in SMMC-7721 cells.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; Apoptosis Inducing Factor ; biosynthesis ; genetics ; Camptothecin ; analogs & derivatives ; pharmacology ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Humans ; Liver Neoplasms ; pathology ; Translocation, Genetic

OBJECTIVETo observe the clinical effect of electric stimulation of long-term retaining needle on analgesia after laparoscopic cholecystectomy (LC) and the impacts on the post-surgical flatus time.

METHODSUnder static absorptive composite general anesthesia, 90 cases of LC were randomized into three groups, 30 cases in each one. In the control group, the analgesia was not applied after LC. In the analgesia-pumper group, the patient controlled intravenous analgesia (PCIA) was used. In the needle-retaining group, the electric acupuncture stimulator was used. The needles were inserted transversely at Riyue (GB 24), Qichong (ST 30) and Yanglingquan (GB 34) and fixed with sterile sticker. Separately, in 8 h and 24 h after surgery, the electric acupuncture stimulation with disperse-dense wave, 2 Hz/100 Hz frequency was applied continuously for 30 min. Visual analogue scale (VAS), adverse reactions such as vomiting and nausea and the postoperative flatus time in 2, 4, 8, 12, 24 and 36 h after surgery were observed and recorded in the three groups.

RESULTSIn 2, 4, 8, 12 and 24 h after surgery, VAS scores in the needle-retaining group and the analgesia-pumper group were all lower than those in the control group (P < 0.05, P < 0.01). The analgesia effect at the above time points in the needle-retaining group was better than that in the analgesia-pumper group (all P < 0.05). There was not adverse reaction in the needle-retaining group. But there were 3 cases of somnolence, 6 cases of nausea and 3 cases of vomiting in the analgesia-pumper group, and 2 cases of nausea and 1 case of vomiting in the control group. The flatus time was quite earlier in the needle-retaining group as compared with the other two groups [(14.77 +/- 4.99) h vs (18.50 +/- 4.22) h, P < 0.01; (14.77 +/- 4.99) h vs (18.17 +/- 4.69) h, P < 0.05].

CONCLUSIONThe electric stimulation of long-term retaining needle is safe and effective in analgesia after LC. It avoids the adverse reactions of analgesics and promotes postoperative flatus.

Acupuncture Analgesia ; instrumentation ; Adult ; Aged ; Cholecystectomy, Laparoscopic ; adverse effects ; Electroacupuncture ; instrumentation ; Female ; Humans ; Male ; Middle Aged ; Pain Management ; Pain, Postoperative ; etiology ; therapy

Objective To elucidate the mutations of NPHS1 gene in children with sporadic steroid-resistant nephrotic syndrome (SRNS) in Southern Chinese Han ethnic group.Methods Peripheral blood samples were collected for genetic analysis from 40 patients with sporadic SRNS and 50 healthy volunteers as control.Genomic DNA was isolated from peripheral blood leucocytes.Twenty-nine exons and exon-intron boundaries of the NPHS1 gene were amplified by polymerase chain reaction.Mutational analysis was performed by DNA sequencing directly.Results Seven variants,928G＞A(D310N),2677A＞G (T893A),2869G＞C (V957L),IVS8+30C＞T,IVS21+14G＞A,IVS25-23C＞T and *142T＞C,of NPHS1 gene were found in 6 of 40 children with sporadic SRNS,whereas they were not found in 50 healthy controls.2677A ＞G,IVS8 +30C ＞T,IVS21 +14G＞A,IVS25-23C ＞T and *142T＞C were novel.Moreover,thirteen already reported NPHS1 polymorphisms,294C＞T,349G＞A,IVS3+15C＞T,IVS3+61A＞G,803G＞A,IVS8+68A＞G,1339G ＞A,1802G ＞C,2223C ＞T,2289C ＞T,IVS24 +36C ＞T,3315G＞A and IVS27 +45C ＞T,were detected in some patients and controls. Conclusions NPHS1 mutations in 6 of 40 children with sporadic SRNS in Southern Chinese Han ethnic group (15％) are detected.NPHS1 mutations are existed in Southern Chinese children,so it is necessary to perform the mutation analysis of NPHS1 gene in those children patients.

Objective To investigate ultrasonographic (US) features of mucinous breast carcinoma with pathological correlation and to improve the early diagnosis for the mucinous breast carcinoma.Methods Twenty-two patients with 23 focuses mucinous carcinoma of the breast confirmed by pathology were included in this study,retrospectively evaluate sonographic features according to the American College of Radiology(ACR) Breast Imaging Reporting and Data System(BI-RADS) Lexicon,all cases were classified by BI-RADS before operation.Histological type of mucinous breast carcinoma included mixed and pure two forms,the latter was classified into cellular variants and hypocellular variants.The correlation between the ultrasonographic findings of mucinous carcinoma and histologic features in different types were analysed.Results On US examination,all 22 cases presented as solid mass.In pure form,they showed well-defined margins with no pseudocapsule and isoechogenic or hypoechoic internal echo pattern relative to that of subcutaneous fat in 86.7％ (13/15) focuses which histological type were pure type.93.3％ (14/15) of pure type focuses had posterior enhancement.All pure type focuses demonstrated that the orientation of the mass is parallel to the skin line.75.0％ (6/8) of mixed type focuses and 13.3％ (2/15) of pure type focuses,however,demonstrated ill-defined and or spiculated margins with more hypoechogenic structure (P ＜0.01).Mixed type focuses have more aggressive malignant features.65.2％ (15/23) of cases being classified as BI-RADS 4 or 5 were suspected as malignancy,which included 6 mixed type focuses and 9 pure type respectively accounting for 75.0％ (6/8) and 60.0％ (6/15) of each type focuses.Conclusions Mucinous breast carcinoma especial pure type did not have the typical malignant breast masses sonographic features,which showed some benign masses sonographic features except no pseudocapsule benign feature.Mixed type mucinous carcinomas tumors have more aggressive margin imaging characteristics.It is suggested that mucinous breast carcinoma should be classified category 4 by BI-RADS to avoid diagnosis delay.

Objective To observe the in vitro anti-Coxsackievirus B3m (CVB3m) effect of sophocarpine(SC) extracted from Sophora flavescens, a traditional Chinese herb. Methods HeLa cells were cultured and the micro-dose cytopathic effect (CPE) assays were applied to detect the toxicity of SC. CPE-inhibitory assays were used to observe the in vitro anti-CVB3m effect of SC. MTT and crystal assays were introduced to examine the anti-CVB3m effect of SC. HeLa cells were infected with CVB3m and added with SC in different concentrations 15 h later.The viability and number of survival of HeLa cells were determined by MTT and crystal violet assays, respectively. Results No toxicity was found on HeLa cells by SC with concentrations 100 ?g/mL, SC could accelerate and aggravate the CPE. SC could protect the CVB3m-infected HeLa cells with concentrations from 1.56 to 25 ?g/mL, and the viability and cell number measured by MTT and crystal violet assay in the SC-handled cells were higher and bigger than those in the virus infected ones. However, the inhibitory effect of virus was exacerbated with higher concentrations (50 and 100 ?g/mL), and the cell number and viability of the SC-handled cells were smaller and lower than those of the infected ones. Conclusion SC with a proper concentration has the in vitro anti-CVB3m effect and may protect HeLa cells from CVB3m infection.