Establishment of Comprehensive Protection System for Intellectual Property of Single Herbs by Taking Callicarpa nudiflora of Jiuzhitang as An Example / 中国实验方剂学杂志

The development of single Chinese materia medica is an important direction of technological innovation in the field of Chinese materia medica at present， and the study of its comprehensive intellectual property protection system is of great significance to the intellectual property protection of the whole chain of innovative enterprises of single Chinese materia medica. Based on this， this paper takes the comprehensive protection system of intellectual property of Callicarpa nudiflora constructed by Jiuzhitang Pharmaceutical as a model to conduct empirical research， analyzes the protection forms applicable to intellectual property of Chinese materia medica， such as patents， administrative protection， trademarks， designs and intangible cultural heritages， and discusses the valuable and insufficient aspects of the protection system currently constructed by Jiuzhitang Pharmaceutical and puts forward the following suggestions：①paying attention to patent applications for planting/processing methods of raw medicinal materials， ②emphasizing the protection of geographical indications， authentic medicinal herbs， and new plant varieties， ③actively promoting product and technology upgrades， ④applying for data protection during product iteration， ⑤emphasizing the layout timing of patent and administrative protection， ⑥focusing on improving goodwill， ⑦enhancing awareness of intellectual property protection and promoting deep integration of industry， academia， and research. We hope that innovative enterprises engaged in the development of single Chinese materia medica can learn from the experience of the case， and optimize the strategy to better protect related products.

CaCO3 blowing agent mixing method for biomass composites improved buffer packaging performance.

Biodegradable composites with an open-cell structure were developed to replace petroleum-based buffer packaging materials. To overcome the problem of uneven and insufficient foam in the composites, CaCO3 was used as a nucleating agent to prepare porous composites. At 5 wt% CaCO3, more uniform and dense composite cells with better cushioning performance were obtained. A further increase in the CaCO3 content caused the density of the cells and the cushioning properties of the composites to decrease. The addition of CaCO3 improved the thermal stability and water barrier properties. The moisture absorption was reduced by 15%. X-ray diffraction analysis indicated that the addition of CaCO3 destroyed the crystalline structure of the starch and produced a new crystalline peak, resulting in a significant reduction in the crystallinity. The decrease in the crystallinity of the starch resulted in the formation of a homogeneous slurry that produced a uniform foam in the composites.

[Effect of electroacupuncture of "Hegu" (LI4) and "Zusanli" (ST36) on intestinal sensitivity and motility in irritable bowel syndrome rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) of "Hegu"(LI4) and "Zusanli"(ST36)on changes of intestinal sensitivity and colonic motility and expression of colonic 5-hydroxytryptamine 3A receptor (5-HT3AR) in irritable bowel syndrome (IBS) rats, so as to reveal its mechanism underlying improvement of IBS. METHODS: A total of 40 neonatal Wistar rats were randomly and equally divided into normal control, model, LI4 and ST36 groups (n＝10). The IBS model was induced by mother-infant separation, acetic acid enema and colorectal distension (CRD). EA (2 Hz/100 Hz, a tolerable strength) was applied to bilateral LI4 and ST36 for 20 min, once every other day for 5 times. The Bristol stool form scale was used to assess the gastrointestinal function, and the latency and number of abdominal muscular contraction waves of abdominal withdrawal reflex (AWR) were used to evaluate the intestinal sensitivity and motility respectively. The immunoactivity of 5-HT3AR of the colon tissue was detected by immunohistochemistry. RESULTS: After modeling, the score of Bristol fecal form scale, number of muscular contraction waves and expression levels of colonic 5-HT3AR in the myometrium and mucosal layers were significantly increased (P<0.01), and the latency of muscular initial contraction wave was obviously shortened in the model group relevant to the normal control group (P<0.01). After the intervention, the increased Bristol fecal form score, number of muscular contraction waves and expression levels of 5-HT3AR in the myometrium and mucosal layers as well as the decreased latency of muscular contraction were reversed in both LI4 and ST36 groups (P<0.01, P<0.05). The effect of EA of ST36 was significantly superior to those of EA-LI4 in lowering Bristol fecal scale score and 5-HT3AR expression in the muscular layer (P<0.01), but obviously inferior to those of EA-LI4 in increasing the latency of of muscular initial contraction wave and down-regulating muscular contraction waves and 5-HT3AR expression in the mucosal layer (P<0.05, P<0.01). CONCLUSION: Both EA-LI4 and EA-ST36 can significantly improve the symptoms of abdominal pain and diarrhea, but EA-LI4 is better in suppressing intestinal high sensitivity, and EA-ST36 is better in promoting intestinal motility, suggesting a specificity of effect of acupoints of different meridians.

[Comparison of effects of electroacupuncture at "Tianshu" (ST25) and "Dachangshu" (BL25) on intestinal sensitivity and expression of muscarinic M3R and 5-HT3AR in irritable bowel syndrome rats].

OBJECTIVE: To observe and compare the effect of electroacupuncture (EA) of "Tianshu" (ST25) and "Dachangshu" (BL25) on intestinal sensitivity and the expression of M3 muscarinic acetylcholine receptor (M3R) and 5-hydroxytryptamine 3A receptor (5-HT3AR) in the colon tissue in irritable bowel syndrome (IBS) rats, so as to explore its mechanism underlying improvement of IBS. METHODS: Forty newborn Wistar rats were randomly divided into blank control, model, ST25 and BL25 groups (n＝10 rats per group). The IBS model was established by joint application of maternal separation, acetic acid enema plus colorectal distension from day 8 to 21 after birth. At the age of 9 weeks, EA (2 Hz/100 Hz, 0.1－0.3 mA) was applied to ST25 and BL25 for 20 min, once every other day for 5 times. The modified method was used to record the abdominal withdrawal reflex (AWR), so as to evaluate the visceral sensitivity by referring to the modified Al-Chaer's and colleagues' methods. The threshold (initial wave), the number and peak-peak value of contraction waves (AWR) within 90 s were recorded. The immunoactivity of M3R and 5-HT3AR in the colon tissue was detected by immunohistochemistry. RESULTS: After modeling, the latency was obviously shortened, the number and peak-peak values of contraction waves of intestine were significantly increased, and the immunoactivity of M3R and 5-HT3AR was notably up-regulated in the model group compared with the blank control group (P<0.01). After the EA intervention, the latency was obviously prolonged, the peak-peak value in both ST25 and BL25 groups, and the number of contraction waves in the ST25 group were significantly decreased (P<0.01), and the expression levels of M3 and 5-HT3A R were considerably down-regulated in both ST25 and BL25 groups relevant to the model group (P<0.01). The therapeutic effects were significantly better in the ST25 group than in the BL25 group in prolonging the latency and reducing the contraction wave number, and in down-regulating the expression of colonic M3R and 5-HT3AR (P<0.05, P<0.01). CONCLUSION: EA at ST25 and BL25 can reduce the intestinal sensitivity in IBS rats, which is probably associated with its effects in down-regulating the expression of M3R and 5-HT3AR in the colon. The therapeutic effects of ST25 were obviously better than those of BL25 in suppressing intestinal sensitivity and expression of colonic M3R and 5-HT3AR.

[Effect of Electroacupuncture at "Neiguan" (PC 6) and "Tianshu" (ST 25) for Colonic Motility and D 2 Receptor in Irritable Bowel Syndrome Rats].

OBJECTIVE: To observe and compare the effects of electroacupuncture (EA) at "Tianshu" (ST 25) and "Neiguan" (PC 6) for colonic motility and the expression of colon dopamine D 2 in irritable bowel syndrome (IBS) rats, and to explore the specificity of different meridians and different acupoints. METHODS: Forty Wistar newborn rats were randomly divided into blank, model, Tianshu and Neiguan groups. Separation of mother and child and acetic acid coloclyster combined with colorectal distension were used to establish IBS model in the model, Tianshu and Neiguan groups. At the age of 9 weeks, EA at bilateral ST 25 and PC 6 were applied in the corresponding groups 5 times, once every other day. After the intervention, the Bristol fecal score, the latent period of abdominal retraction reflex and the number of contraction waves were recorded. The expression of dopamine D 2 receptor was detected by immunohistochemistry. RESULTS: Compared with the blank group, the Bristol fecal score of the model group was higher (P<0.01), the 1st contraction wave latent period was shorter (P<0.01), the number of contraction waves in 90 s increased (P<0.01), the immunoreactive expression of D 2 receptor in colon decreased (P<0.01). Compared with the model group, the Bristol fecal scores of the Tianshu and Neiguan groups decreased (P<0.01), the 1st contraction wave latent periods were longer (P<0.01), the numbers of contraction waves in 90 s decreased (P<0.01), the positive expressions of D 2 receptor in colon increased (P<0.01, P<0.05). Compared with the Tianshu group, the immunoreactive expression of D 2 receptor in the Neiguan group decreased (P<0.01). CONCLUSION: EA at ST 25 and PC 6 can improve the symptoms of colonic motility in IBS rats. The effect of EA at ST 25 is better, which indicates that different meridians and different acupoints play specific effects.

[Molluscicidal effect of simple black film mulching in mountainous areas of Yunnan Province].

OBJECTIVE: To evaluate the molluscicidal effect of black plastic mulching in mountainous areas of Yunnan Province. METHODS: In Leqiu Village, Nanjian County, Yunnan Province, the terraced fields with Oncomelania hupensis snails were divided into A, B, C, D, E groups, and of which, A, B, C, D groups, as the experimental groups, were administered with simple black plastic mulching for 30 days, 60 days, 90 days and 120 days respectively; Group E, as the control group, was administered with 50% niclosamide ethanolamine salt wettable powder spraying. RESULTS: In Group C and D (simple black film mulching for 90 days and 120 days), no snails were found after the administration, the average density of living snails decreased by 100%, and the cost of one year was similar to that of the control group. CONCLUSION: The molluscicidal effect of black plastic mulching is good in mountainous areas.

[Investigation on infection sources of schistosomiasis in mountainous regions, Nanjian County, Yunnan Province].

OBJECTIVE: To understand the types of schisotsome infection sources and their roles in schistosomiasis transmission in mountainous endemic regions in Yunnan Province, so as to provide the evidence for implementing the comprehensive control measures based on infection source control. METHODS: Two villages of typical mountainous regions in Nanjian County were chosen for field investigation. The Oncomelania hupensis snail status was surveyed by the methods of systematic and environmental sampling. The infections of schistosomiasis were surveyed in residents with the indirect haemagglutination and the hatching method, in livestock with the hatching method and in wild animals with the anatomical method and the hatching method. The distribution and pollution status of wild faeces were investigated in the snail environments nearby villages or with livestock frequent activities. The positives of schistosomiasis in wild faeces were tested with the hatching method. The pollution index of wild feces was calculated. RESULTS: A total of 533.56 hm2 were investigated in two villages, and the area with snails was 16.52 hm2. The rate of frame with snails was 1.03%, the average density of snails was 0.07 snails/0.1 m2, and no positive snails were found. The positive rate of blood examinations of population was 1.61%, but no persons were positive with the hatching method. The infection rates were 0.90%, and 0.62% in cattle and equus, respectively, and there were no positives in other livestock. Totally 472 piles of wild feces of 6 species (human, cattle, equus, goat, pig and dog) distributed in the investigation areas, and among them, the densities of wild feces of cattle, equus, and dog were 7.2, 4.3 piles/hm2, and 2.1 piles/hm2 respectively, being relatively higher than others. The hatching positive rates of wild faeces of cattle, equus, and dog were 2.27% (3/132), 2.63% (2/76), and 3.70% (1/27), respectively. The relative pollution indexes of wild faeces of cattle, equus, and dog were 80.68%, 15.89%, and 3.43%, respectively. CONCLUSIONS: After schistosomiasis transmission is controlled, the potential epidemic factors still remain in the mountainous endemic regions of schistosomiasis in Yunnan Province. The cattle are still the most infection source of schistosomiasis, but equus and dog as infection sources should be not neglected. We should extend the investigation and monitor scope of the infection sources, and carry out scientific and feasible control technique and management measures.

TAT-CC fusion protein depresses the oncogenicity of BCR-ABL in vitro and in vivo through interrupting its oligomerization.

Chronic myeloid leukemia (CML) is a clonal hematologic malignancy characterized by the BCR-ABL protein. BCR-ABL is a constitutively active tyrosine kinase and plays a critical role in the pathogenesis of CML. Imatinib mesylate, a selective tyrosine kinase inhibitor, is effective in CML, but drug resistance and relapse occur. The coiled-coil (CC) domain located in BCR(1-72) mediates BCR-ABL tetramerization, which is essential for the activation of tyrosine kinase and transformation potential of BCR-ABL. CC domain is supposed to be a therapeutic target for CML. We purified a TAT-CC protein competively binding with the endogenous CC domain to reduce BCR-ABL kinase activity. We found that TAT-CC co-located and interacted with BCR-ABL in Ba/F3-p210 and K562 cells. It induced apoptosis and inhibited proliferation in these cells. It increased the sensitivity of these cells to imatinib and reduced the phosphorylation of BCR-ABL, CRKL and STAT5. We confirmed that TAT-CC could attenuate the oncogenicity of Ba/F3-p210 cells and diminish the volume of K562 solid tumor in mice. We conclude targeting the CC may provide a complementary therapy to inhibit BCR-ABL oncogenicity.

[Effect of snail control of niclosamide by soil mixing and spraying methods in high dam of terrace].

The snail control efffect of niclosamide by soil mixing and spraying method were compared, and the results showed that the effects of the two methods were similar. Though there was difficulties in getting soil and mixing durg with soil, and the cost was high, the effect of soil mixing method in some special environment such as stone ditches and terraces.

[Effect of comprehensive control project on schistosomiasis in Nanjian county].

The comprehensive control project for schistosomiasis was implemented in Nanjian County from 2004 to 2008. After the implementation of the control project, the infection rates of population and livestock decreased by 94.39% and 83.29% in 2008, respectively, with both infection rates less than 1%, no acute schistosomiasis cases had been found since 2005. Snail areas decreased by 70.01%, no infected snails had been found since 2007. Through the implementation of the comprehensive control project, schistosomiasis had been effectively controlled in Nanjian County.

Glycoprotein biosynthesis in porcine aortic endothelial cells and changes in the apoptotic cell population.

Porcine aortic endothelial cells (PAECs) produce glycoproteins with important biological functions, such as the control of cell adhesion, blood clotting, blood pressure, the immune system, and apoptosis. Cell surface glycoproteins play important roles in these biological activities. To understand the control of cell surface glycosylation, we elucidated biosynthetic pathways leading to N- and O-glycans in PAECs. Based on the enzyme activities, PAECs should be rich in complex biantennary N-glycans. In addition, the enzymes synthesizing complex O-glycans with core 1 and 2 structures are present in PAECs. The first enzyme of the O-glycosylation pathway, polypeptide GalNAc-transferase, was particularly active. Its specificity toward synthetic peptide substrates was found to be similar to that of purified bovine colostrum enzyme T1. A significant fraction of PAECs treated with tumour necrosis factor alpha or human serum detached from the culture plate, and most of these cells were apoptotic. The apoptotic cell population exhibited decreased core 2 beta 6-GlcNAc-transferase activity. In contrast, the activities of core 1 beta 3-Gal-transferase, which synthesizes O-glycan core 1, and of alpha 3-sialyltransferase (O), which sialylates core 1, were increased in apoptotic PAECs. Thus, apoptotic PAECs are predicted to have fewer complex O-glycans and a higher proportion of short, sialylated core 1 chains.