A PCR method was used to amplify the sequence encoding the mature peptide of?-mannanase of Bacillus subtilis. The gene was inserted into the Pichia pastoris vector pPIC9K, downstream of?-factor signal peptide sequence. The resultant recombinant plasmid pPIC9K-MAN was lineared by BglII digestion and introduced into the host Pichia pastoris GS115 by PEG method. After screen, the recombinant P. pastoris strain MAN22 was obtained and fermented in large scale 5L fermenter. The recombinant mannanase activity could reach to 1102IU/ml. The properties of the recombinant mannanase were characterized.

OBJECTIVE: To establish an HPLC method for determination of ferulic acid in Ligusticum Wallichii,which is the active ingredient of Xinmaishu capsules. METHODS: The content was determined by HPLC. The analysis was carried out on C18(4.6 mm×250 mm,5 μm) column. The mobile phase consisted of acetonitrile-0.3% acetic acid(13∶87). The flow rate was 1.0 mL·min-1, and the UV detection wavelength was set at 322 nm. RESULTS: The linearity range of the calibration curve for ferulic acid was from 1.0 to 10.0 μg·mL-1 (r=0.999 8). The average recovery rate was 99.57% with RSD of 0.75%(n=9). CONCLUSION: This method is sensitive, accurate,reproducible and simple. It can be used for the quality control of ferulic acid in Xinmaishu capsules.

Objective To evaluate the nutritional status of iodine of pregnant and lactating women in coastal and inland areas of Zhejiang province,and to provide scientific basis for iodine supplementation.Methods In 2009,five coastal cities,five coastal counties and three inland areas from Zhejiang province were selected by the cluster sampling method,and one sub-district(township) was randomly selected in each city (county),then one community (village) was randomly selected in each sub-district (township).Thirty pregnant women,thirty lactating women and fifty children aged 8 - 10,were randomly selected in each community(village) as the investigation objects to collect instant urine samples to detect urine iodine,and to collect thirty resident edible salt and some water samples to detect salt iodine and water iodine,if the numbers of water source were more than 5 then 1 water sample was collected by their location of east,south,west,north and center in each community(village),otherwise,the numbers of water samples collected were equal to the number of community (village).Results The medians of salt iodine of coastal city,coastal county and inland area were 28.42,25.29,and 33.24 mg/kg,respectively,and the medians of water iodine correspondingly were 5.41,11.52,and 2.80 μg/L,respectively.The medians of urinary iodine of pregnant and lactating women in Zhejiang province were 148.72 and 161.34 μg/L,respectively,which were less than that of children aged 8 - 10(231.02 μg/L,Z =8.57,8.24,all P ＜ 0.017).The medians of urinary iodine in pregnant women from different areas were ranked in order as inland areas(211.42 μg/L),coastal counties (150.51 μg/L) and coastal cities (123.33 μg/L,Z =2.80,5.31,2.42,all P ＜ 0.017),and in lactating women,the medians of urinary iodine in inland areas( 197.43 μg/L) was higher than that in coastal cities( 139.64 μg/L,Z =4.03,P ＜ 0.017).Conclusions The overall levels of iodine nutrition in lactating and pregnant women in Zhejiang province are in the appropriate level,but that of the pregnant women in the coastal cities is inadequate.

Good medicinal herbs, good drugs. Good evaluation method and indices are the prerequisite of good medicinal herbs. However, there exist numerous indices for quality evaluation and control in Chinese medicinal materials. However, most of these indices are non-interrelated each other, as well as having little relationship with efficiency and safety. The results of different evaluatior methods may not be consistent, even contradictory. Considering the complex material properties of Chinese medicinal materials, single method and index is difficult to objectively and comprehensively reflect the quality. Therefore, it is essential to explore the integrated evaluation methods. In this paper, oriented by the integrated evaluation strategies for traditional Chinese medicine quality, a new method called integrated quality index (IQI) by the integration of empirical evaluation, chemical evaluation, and biological evaluation was proposed. In addition, a study case of hypertoxic herb Aconitum carmichaelii Debx. was provided to explain this method in detail. The results suggested that in the view of specifications, the average weight of Jiangyou aconite was the greatest, followed by Weishan aconite, Butuo aconite, Hanzhong aconite, and Anxian aconite; from the point of chemical components, Jiangyou aconite had the characteristic with strong efficacy and weak toxicity, next was Hanzhong aconite, Butuo aconite, Weishan aconite, and Anxian aconite; taking toxicity price as the index, Hanzhong aconite and Jiangyou aconite have the lower toxicity, while Butuo aconite, Weishan aconite, and Anxian aconite have the relatively higher one. After the normalization and integration of evaluation results, we calculated the IQI value of Jiangyou aconite, Hanzhong aconite, Butuo aconite, Weishan aconite, and Anxian aconite were 0.842 +/- 0.091, 0.597 +/- 0.047, 0.442 +/- 0.033, 0.454 +/- 0.038, 0.170 +/- 0.021, respectively. The quality of Jiangyou aconite is significantly better than the others (P < 0.05) followed by Hanzhong aconite, which is consistent with the traditional understanding of genuineness. It can be concluded that IQI achieves the integrated control and evaluation for the quality of Chinese medicinal materials, and it is an exploration for building the good medicinal herbs standards. In addition, IQI provides technical supports for the geoherbalism evaluation, selective breeding, the development of precision decoction pieces, high quality and favourable price in market circulation, and rational drug use.
Aconitum ; chemistry ; Drugs, Chinese Herbal ; standards ; Medicine, Chinese Traditional ; standards

The stable and controllable quality of decoction pieces is an important factor to ensure the efficacy of clinical medicine. Considering the dilemma that the existing standardization of processing mode cannot effectively eliminate the variability of quality raw ingredients, and ensure the stability between different batches, we first propose a new strategy for Chinese medicine processing technologies that coupled with individuation processed and cybernetics. In order to explain this thinking, an individual study case about different grades aconite is provided. We hope this strategy could better serve for clinical medicine, and promote the inheritance and innovation of Chinese medicine processing skills and theories.
Chemistry, Pharmaceutical ; methods ; standards ; Cybernetics ; standards ; Drug Therapy ; standards ; Drugs, Chinese Herbal ; chemistry ; standards ; toxicity ; Humans ; Quality Control

Quality control is one of the key scientific tissues in the modernization of traditional Chinese medicines (TCM). In order to overcome the deficiencies of assessment indexes, including little systematization and quantification, as well as loose association with clinical efficacy and dosage, a new integrated method named controllability pyramid ( CP) is first proposed in this paper. In addition, some study cases are used to explain how this model is constructed. We hope the establishment of CP could promote the clinical-orien- ted integrated innovation research of TCM, and provide control strategy and technology examples for improving the quality of Chinese medicines and clinical efficacy.
China ; Drug Evaluation ; methods ; standards ; Drug Therapy ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; standards ; Humans ; Medicine, Chinese Traditional ; standards ; Quality Control

OBJECTIVETo compare the therapeutic differences among scalp acupuncture combined with auricular point sticking, body acupuncture and western medication for treatment of vascular dementia (VD).

METHODSNinety cases were randomly divided into a combined therapy group (31 cases), a body acupuncture group (29 cases) and a western medication group (30 cases). The combined therapy group was treated with scalp acupuncture at forehead middle line, parieral middle line, temporal front line and temporal rear line as well as auricular point sticking at naogan (AT3,41), shen (CO10), shenmen (TF4), zhen (AT3), once a day; the body acupuncture group was treated with acupuncture at Baihui (GV 20), Fengchi (GB 20), Zusanli (ST 36) and so on, once a day; the western medication group was treated with oral administration of aniracetam tablets, 0.2 g per time, twice a day. Fourteen days were considered as a treatment course, and totally 3 courses were required. The mini-mental state examination (MMSE) and activities of daily living (ADL) were applied to assess the changes of cognitive behavior ability before and after treatment among three groups. Also the efficacy among three groups were compared.

RESULTSOne case dropped out in the body acupuncture group and western medication group, respectively. The total effective rate was 90.334 (28/31) in the combined therapy group, which was superior to 85.734 (24/28) in the body acupuncture group and 79.3% (23/29) in the western medication group (both P < 0.05). After the treatment, the scores of MMSE and ADL were all improved among three groups, which was the most significant in the combined therapy group (MMSE: 23.32 +/- 4.45 vs 21.23 +/- 4.13, P < 0.05; 23.32 +/- 4.45 vs 20.41 +/- 4. 01, P < 0.01; ADL: 53.18 +/- 21.55 vs 51.92 +/- 20.42, P < 0.05; 53.18 +/- 21.55 vs 49.42 +/- 19.43, P < 0.01).

CONCLUSIONThe scalp acupuncture combined with auricular point sticking could improve the clinical symptoms and cognitive behavior ability in patients with vascular dementia, which has superior total efficacy to body acupuncture and western medication aniracetam tablets.

Activities of Daily Living ; Acupuncture Points ; Acupuncture Therapy ; Aged ; Cognition ; Dementia, Vascular ; psychology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Scalp

OBJECTIVETo study COX-2 expression in H. pylori infected gastric mucosal epithelia and its significance in the carcinogenesis of the stomach.

METHODSRapid urease test and histological examination with basic magnenta staining were used to assess the status of H. pylori infection in the stomach. COX-2 was detected immunohistochemically.

RESULTSCOX-2 immunostaining was positive in 1 out of 12 cases with H. pylori-negative gastric mucosa and also in 1 out of 10 cases with H. pylori-positive gastric mucosa without macroscopic alterations, while COX-2 expression was found to be positive in 5 out of 9 cases with H. pylori related superficial gastritis with mucosal erosions. COX-2 expression was detected in 5 out of 10 cases with H. pylori-positive mild atrophic gastritis, 8 out of 10 cases with H. pylori-positive moderate-severe atrophic gastritis and intestinal metaplasia, and 6 out of 8 cases with H. pylori-positive moderate-severe dysplasia. COX-2 expression was positive in 22 out of 32 cases of gastric cancer.

CONCLUSIONH. pylori may induce COX-2 expression of gastric mucosal epithelia in chronic superficial gastritis, which is related to the development of mucosal injury. According to gastric mucosal carcinogenesis pattern up-regulation of COX-2 expression is associated with gastric mucosal carcinogenesis, and involved in the early development of premalignant lesions.

Adult ; Aged ; Cyclooxygenase 2 ; biosynthesis ; genetics ; Female ; Gastric Mucosa ; enzymology ; Gastritis ; enzymology ; microbiology ; Helicobacter Infections ; enzymology ; Helicobacter pylori ; Humans ; Male ; Middle Aged

Objective To evaluate the dietary iodine intake level of population in coastal and inland areas of Zhejiang province.Methods The cluster sampling method was applied to select Zhoushan,Ningbo and Taizhou cities from Zhejiang province as coastal areas,to select Jinhua,Quzhou and Lishui cities from Zhejiang province as inland areas,and two counties(districts) were randomly selected in each chosen city,three sub-districts(towns) were randomly selected in each chosen counties (districts),then one community (village) was randomly selected in each chosen sub-districts(towns).Adult residents aged greater than 18 were selected as investigation subjects,their dietary iodine intake in the past 24 hours was investigated,and the Food composition table” was inquired to get the data of dietary iodine intake.One hundred copies of residential edible salt samples and drinking water samples in each of the selected community (village) were collected to detect salt iodine and water iodine by direct titrimetric and spectrophotometric method,respectively.Results The mean of adult residents' dietary iodine intake in Zhejiang coastal area was 267.76 μg/d,which was less than that in inland area (429.05 μg/d,t =- 6.90,P ＜0.05),and the means of adult residents' dietary iodine intake from drinking water,laver and fish(5.75,69.72,5.61 μg/d,respectively) in coastal area were higher than those in inland areas(3.25,35.27,3.43 μg/d,respectively,t =21.73,3.92,4.08,all P ＜ 0.05),however,the means of adult residents' dietary iodine intake from salt,kelp and other food (166.81,3.04,16.82 μg/d,respectively) in coastal areas were less than those in inland areas (355.15,6.14,25.81 μg/d,respectively,t =- 8.76,- 5.49,- 18.56,all P ＜ 0.05).In coastal areas,the proportion which was less than estimated average requirement of iodine(EAR,120 μg/d) was 46.48％ (1029/2214),the ratio which was higher than the maximum tolerable intake of iodine(UL,1000 μg/d) was 3.34％ (74/2214),and the average contribution rate of dietary intake of iodine in salt was 62.30％(166.81/267.76).While in inland areas,the corresponding proportions were 7.61％ (171/2246),2.80％ (63/2246) and 82.78％ (355.15/429.05),respectively.Conclusions The dietary iodine intake in Zhejiang inland areas has reached the recommended nutrient intake levels of the Chinese Nutrition Society,but there is a certain degree of insufficient iodine intake in population of the coastal areas.

Humans ; Laryngeal Neoplasms ; pathology ; Male ; Middle Aged ; Neurilemmoma ; pathology