Sini Decoction combined with spectinomycin B1 induced dissociation of PTEN and SUMO1 to treat lung cancer / 中草药

Objective To analyze the mechanism of the inhibition of proliferation, metastasis and the promotion of apoptosis of lung cancer cells by Sini Decoction combined with spectinomycin B1 from the perspective of protein SUMOylation modification. Methods The experiment was divided into control group, spectinomycin B1 group, Sini Decoction group, and combined treatment group. Human non-small cell lung cancer cell line A549 was treated for 48 h and Western blotting was used to detect the protein expression of small ubiquitin-related modified protein-1 (SUMO1), gene of phosphate and tension homology deleted on chromsome ten (PTEN), and phosphorylated Akt (p-Akt) and Caspase-9. Cell proliferation curves were drawn by MTT assay, and cell apoptosis was detected by flow cytometry; Cell migration and cell invasion were detected by cell scratch and Transwell assay. Results Both spectinomycin B1 and Sini Decoction an reduce the levels of covalent SUMO1 and p-Akt, increase the protein levels of PTEN and activate Caspase-9, inhibit cell proliferation, migration and invasion, and promote cell apoptosis. Combined treatment group can further enhance the above changes. Conclusion Sini Decoction can synergize with spectinomycin B1 to induce the dissociation of PTEN and SUMO1, thereby inhibiting the proliferation, metastasis and promoting apoptosis of lung cancer cells.

Determination of Related Substances of Spectinomycin Hydrochloride for Injection by Two Analysis Methods / 中国药学杂志

OBJECTIVE: To compare the HPLC-PAD method and HPLC-ELSD method for determination of related substances in spectinomycin hydrochloride for injection, and determine 31 batches of samples by the two methods. METHODS: The following conditions were used for both methods: the analytical column was Apollo C18 column (4.6 mm×250 mm, 5 μm), mobile phase was 0.1 mol·L-1 trifluoroacetic acid solution, the column temperature was maintained at 30 ℃, and the sample volume was 20 μL. For the PAD detection, gold electrode (3 mm)was used as the detection electrode, Ag-AgCl as reference electrode, titanium alloy as counter electrode, and integrated amperometric detector was employed with four waveform detection potential. For the ELSD detection, the drift tube temperature was set at 110℃, and the carrier gas flow rate was 2.6 L·min-1, with gain of 1. RESULTS: The two methods were consistent in the species and number of detected impurities. The detection limits of the PAD method and ELSD method were 2.4 and 72.8 ng, respectively (S/N=3). For the PAD method there existed a direct linear relationship in the concentration range of 0.15-150 μg·mL-1, while a double logarithmic linear relationship was shown in the concentration range of 17-170 μg·mL-1 for the ELSD method. When 31 batches of samples were determined for related substances by the two kinds of methods, the contents of impurity D and E were basically the same, but there were significant differences in the contents of impurity A and (4R)-dihydrospectinomycin. After correction, the contents of impurity A and total impurities determined by the PAD method were consistent with those by the ELSD method. CONCLUSION: The two methods can both effectively detect the related substances of spectinomycin to achieve the purpose of controlling related substances. When using HPLC-PAD method, the impurity A and (4R)-dihydrospectinomycin should be adjusted by using response factor or the reference substances. The ELSD method needs to use double log linear regression.

Mutations in the rpsE gene and spectinomycin resistance in Neisseria gonorrhoeae / 中华皮肤科杂志

Objective To evaluate the relationship between spectinomycin resistance in Neisseria gonorrhoeae and mutations in the rpsE gene.Methods Genomic DNA was extracted from 4 clinical isolates of Neisseria gonorrhoeae with different levels of spectinomycin resistance.Then,PCR was performed to amplify the entire rpsE gene and the spectinomycin resistance-determining region (SRDR) in the 16S rRNA gene followed by direct sequencing.Two spectinomycin-sensitive Neisseria gonorrhoeae strains were transformed with the genomic DNA containing the mutant rpsE gene.Subsequently,the susceptibility of the transformants to spectinomycin was determined,and PCR was performed to amplify the rpsE and 16S rRNA genes in the transformants followed by sequencing.Results All the 4 spectinomycin-resistant Neisseria gonorrhoeae strains harbored an A70C transversion in the rpsE gene,but no abnormality in the SRDR of the 16S rRNA gene.No mutations were detected in the spectinomycin-sensitive Neisseria gonorrhoeae strains.The A70C transversion in the rpsE gene was also detected in the two Neisseria gonorrhoeae transformants with spectinomycin resistance.Conclusion The A70C point mutation within the rpsE gene is associated with spectinomycin resistance in Neisseria gonorrhoeae.

Surveillance on Antibiotic Susceptibility of Neisseria Gonorrhoeae from 1988 to 2002 in Shanghai / 中华皮肤科杂志

Objective To study the susceptibility of Neisseria gonorrhoeae to antibiotic agents from 1988 to 2002 in Shanghai. Methods The clinical isolates from patients with gonorrhea were collected and tested for their susceptibility to five antibiotics. Agarose-dilution-method was used to detect minimal inhibitory concentration (MIC) of anti-microbial agents including penicillin, tetracycline, spectinomycin, ciprofloxacin and ceftriaxone, and penicillinase producing Neisseria gonorrhoeae (PPNG) were tested with acidometric method. Results Susceptible strains to penicillin decreased from 11.28% in 1988 to 0 in 2002, MIC50 and MIC90 increased 8 and 4 times, respectively, the resistant rate and proportion of PPNG were 94.29% and 50.95%, respectively in 2002. The strains of high resistance to tetracycline increased from 0 in 1995 to 20.95% in 2002. The susceptible strains to ceftriaxone decreased from 100% in 1995 to 23.80% in 2002. The susceptibility to ciprofloxacin decreased significantly and resistant rate reached 99.05% in 2002. However, these strains were kept highly susceptible to spectionmycin. Concerning the multi-drug resistance, we found that the strains resistant to penicillin, ciprofloxacin and tetracycline simultaneously increased from 20.87% in 2001 to 23.30% in 2002, those resistant to both penicillin and ciprofloxacin reached to 70% in the past 2 years. Conclusions In Shanghai the resistant rates of Neisseria gonorrhoeae to antibiotics increased year by year in the past 15 years. The study indicates that spectinomycin and ceftriaxone should be the first choice for the treatment of gonorrhea at present and new sensitive antibiotic should be developed for the treatment of gonorrhea.

Antimicrobial Susceptibility of Mycoplasma and Clinical Response to Antimicrobial Agents(Spectinomycin etc.)in Genitourinary Mycoplasma Infection / 中华皮肤科杂志

Objective To investigate the antimicrobial susceptibility of spectinomycin?minocycline?azithromycin and sparfloxacin to mycoplasma(Uu and Mh)and therapeutic effect of spectinomycin to my-coplasma infection in genitourinary tract.Methods①The susceptibility test:each of the4drugs was divided into two concentrations.One was at1?g/mL(sensitive concentration)and the other was at4?g/mL(resistant concentration).If mycoplasma does not grow in both concentrations,it means the drug tested is sensitive.If it grows in both concentrations,the drug tested is resistant.If mycoplasma grows in lower concentration and does not in higher concentration,it means moderate sensitive.②Treatment regimen:Spectinomycin was injected,2g/d IM,for7-10days as a course of treatmeant.Patients were followed-up7days later and2~4weeks after treatment.Results①Among1658specimens,519were found Uu positive,and61Mh positive.The resis-tance rates of Uu to4different drugs were:7.7%for minocycline,21.4%for sparfloxacin,13.9%for azithromycin and7.3%for spectinomycin.Whereas,those of Mh were:18.0%,45.9%,54.1%,and29.5%re-spectively.②The clinical effect of spectinomycin was:out of43treated patients,37(86.0%)cured,4(9.3%)markedly improved,2(4.7%)failed.Total effective rate was95.3%and so was the elimination rate of my-coplasma.Conclusion The resistant rate of mycoplasma to spectinomycin is lower than that to minocycline?azithromycin and sparfloxacin,and the former is widely used in the treatment of mycoplasma(especially Uu)infection,with a satisfactory clinical effect.

Treatment of Gonorrhoea

Because of increasing resistance of circulating N. gonorrhoeae and frequent failures in the treatment of gonorrhoea, intensive work on gonorrhoea has become of paramount importance. During January 1980-April 1984, at the Choong-Ku VD Clinic in Seoul, 3,340 male patients with uncomplicated gonococcal urethritis were treated with various treatment regimens. Diagnosis of gonorrhoea and declaration of a treatment failure were made on the basis of positive urethral culture. In 1984, the prevalence of Penicillinase Producing N. gonorrhoeae (PPNG) was about 30%, The pretreatment minimun inhibitory concentration of various antibiotics were quite high. Even for non-PPNG urethritis standard penicillin regimens gave unsatisfactory results. For PPNG urethritis, only spectinomycin, cefoperazone and cefotaxim-probenecid regimens gave satisfactory results. No spectmomycin resistant strain of N. gonorrhoeae has been found since 1982 at the Choong-Ku VD Clinic. As an agent of single drug therapy, spectinomycin seems to be one of the most cost effective drugs in the treatment of uncomplicated gonorrhoea in men.

IN VITRO ANTIBACTERIAL ACTIVITIES OF FOUR ANTIBACTERIAL AGENTS TO NEISSERIA GONORRHEAE / 重庆医科大学学报

In vitro susceptibilities of 38 strains of gonococcus to 4 antibacterial agents wers tested by the agar double dilution method.Beta-lactamase of the bacteria was also tested.The rusults showed that in 38 strains of gonococcus 21.1% were penicillin resistant, 7.9% were PPNG strains, 7.9%were spectinomycin resistant,all of them were susceptible to both ofloxalin and norfloxacin.The MIC50 and MIC90 of ofloxacin and norfloxacin were very low.It was indicated thaat much attention should be paid to the resistance of gorcccccus to penicillin and spectinomycin. Ofloxacin and norfloxacin should be recommended in treatment of gonorrhea

Clinical Effect of Spectinomycin on Acute Gonorrheal Urethritis / 대한비뇨기과학회지

Forty five gonococcal patients were treated with spectinomycin hydrochloride at the Department of Urology, St. Mary's Hospital from August 1. 1972 to December 32. 1972. All patients were male and confirmed as acute gonorrheal urethritis by gram stained smear or culture. The cure ability in this series was very satisfactory The results were as follows; 1. Gram negative ;intracellular diplococci was controlled in 29 patients (96.7%) among 3O patients with pure gonococcal infection with a single injection of spectinomycin hydrochloride. 2. Gram negative intracellular diplococci was controlled in 14 patients (93.3%) among 15 patient with mixed infection with a single injection of spectinomycin hydrochloride. 3. Only two in all patients were controlled in gram negative intracellular diplococci with repeat injection. 4. 4 patients (13.3%) of the pure gonoccoccal patients with a single injection were converted to non-specific urethritis, which were treated with combination of other antibiotics. 5. 5 patients (33.3%) of the mixed infected patients with a single injection were converted to nonspecific urethritis, which were treated with combination of other antibiotics. 6. No side effect was noted in all patients.

Clinical Effect of Spectinomycin on Acute Gonorrheal Urethritis / 대한비뇨기과학회지

Forty five gonococcal patients were treated with spectinomycin hydrochloride at the Department of Urology, St. Mary's Hospital from August 1. 1972 to December 32. 1972. All patients were male and confirmed as acute gonorrheal urethritis by gram stained smear or culture. The cure ability in this series was very satisfactory The results were as follows; 1. Gram negative ;intracellular diplococci was controlled in 29 patients (96.7%) among 3O patients with pure gonococcal infection with a single injection of spectinomycin hydrochloride. 2. Gram negative intracellular diplococci was controlled in 14 patients (93.3%) among 15 patient with mixed infection with a single injection of spectinomycin hydrochloride. 3. Only two in all patients were controlled in gram negative intracellular diplococci with repeat injection. 4. 4 patients (13.3%) of the pure gonoccoccal patients with a single injection were converted to non-specific urethritis, which were treated with combination of other antibiotics. 5. 5 patients (33.3%) of the mixed infected patients with a single injection were converted to nonspecific urethritis, which were treated with combination of other antibiotics. 6. No side effect was noted in all patients.