[Effect of Liangfang Wenjing Decoction on expression of key glycolytic enzymes in uterus and ovaries of rats with coagulating cold and blood stasis syndrome].

This study aimed to investigate the relationship between coagulating cold and blood stasis syndrome and glycolysis, and observe the intervention effect of Liangfang Wenjing Decoction(LFWJD) on the expression of key glycolytic enzymes in the uterus and ovaries of rats with coagulating cold and blood stasis. The rat model of coagulating cold and blood stasis syndrome was established by ice-water bath. After modeling, the quantitative scoring of symptoms were performed, and according to the scoring results, the rats were randomly divided into a model group and LFWJD low-, medium-and high-dose groups(4.7, 9.4, 18.8 g·kg~(-1)·d~(-1)), with 10 in each group. Another 10 rats were selected as the blank group. After 4 weeks of continuous administration by gavage, the quantitative scoring of symptoms was repeated. Laser speckle flowgraphy was used to detect the changes of microcirculation in the ears and uterus of rats in each group. Hematoxylin-eosin(HE) staining was used to observe the pathological morphology of uterus and ovaries of rats in each group. The mRNA and protein expressions of pyruvate dehydrogenase kinase 1(PDK1), hexokinase 2(HK2) and lactate dehydrogenase A(LDHA) in the uterus and ovaries of rats were examined by real-time quantitative polymerase chain reaction(RT-qPCR) and Western blot, respectively. The rats in the model group showed signs of coagulating cold and blood stasis syndrome, such as curl-up, less movement, thickened veins under the tongue, and reduced blood perfusion in the microcirculation of the ears and uterus, and HE staining revealed a thinning of the endometrium with disorganized arrangement of epithelial cells and a decrease in the number of ovarian follicles. Compared with the model group, the treatment groups had alleviated coagulating cold and blood stasis, which was manifested as red tongue, reduced nail swelling, no blood stasis at the tail end as well as increased blood perfusion of the microcirculation in the ears and uterus(P<0.05 or P<0.01). Among the groups, the LFWJD medium-and high-dose groups had the most significant improvement in coagulating cold and blood stasis, with neatly arranged columnar epithelial cells in uterus, and the number of ovarian follicles was higher than that in the model group, especially mature follicles. The mRNA and protein expressions of PDK1, HK2, LDHA in uterus and ovaries were up-regulated in the model group(P<0.05 or P<0.01), while down-regulated in LFWJD medium-and high-dose groups(P<0.05 or P<0.01). The LFWJD low-dose group presented a decrease in the mRNA expressions of PDK1, HK2 and LDHA in uterus and ovaries as well as in the protein expressions of HK2 and LDHA in uterus and HK2 and PDK1 in ovaries(P<0.05 or P<0.01). The therapeutic mechanism of LFWJD against coagulating cold and blood stasis syndrome is related to the down-regulation of key glycolytic enzymes PDK1, HK2 and LDHA, and the inhibition of glycolytic activities in uterus and ovaries.

Effects of cullin 4B on the proliferation and invasion of human gastric cancer cells.

The major aim of the present study was to explore the effects of cullin 4B (CUL4B) on the proliferation and invasion of human gastric cancer cells. Gastric tumor tissues and paired adjacent non­tumor tissues were obtained from 21 gastric cancer patients, and gastric cancer cell lines (AGS, MGC­803, KATO­III, MKN­45, SGC­7901, BGC­823 and MKN­74) were cultured. BGC­823 cells were transfected with CUL4B small interfering (si)RNA or control siRNA. Reverse transcription­quantitative polymerase chain reaction analysis was performed to detect the mRNA expression of CUL4B. Western blot analysis was performed to measure the protein levels of Wnt, ß­catenin, glutathione synthase kinase (GSK)­3ß, caspase­3 and cyclin E. MTT and Transwell assays were performed to examine cell proliferation and invasion following CUL4B knockdown. In addition, the effect of CUL4B knockdown on the cell cycle and apoptosis of BGC­823 cells was evaluated by flow cytometric analysis. The results indicated that compared with the adjacent non­tumor tissues and a normal gastric epithelial cell line, gastric cancer tissues and cell lines exhibited significantly higher expression of CUL4B. Knockdown of CUL4B in gastric cancer cells suppressed cell proliferation, caused G1 arrest and inhibited cell invasion. Silencing of CUL4B also resulted in decreased Wnt and ß­catenin expression, but increased expression of GSK­3ß, caspase­3 and cyclin E. These results indirectly demonstrate that CUL4B enhances the proliferation and invasion abilities of gastric cancer cells by upregulating the constituent factors Wnt and ß­catenin, as well as by negatively regulating the mRNA and protein expression of GSK­3ß, caspase­3 and cyclin E. The potential mechanism of CUL4B highlighted in the present study may be helpful for the treatment of patients with gastric cancer.

Effects of Modified Wenjing Decoction on the ovarian oxidative damage of cold coagulation blood stasis model rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effects of Modified Wenjing Decoction (MWJD) on the ovarian oxidative damage of cold coagulation blood stasis (CCBS) model rats, thus consummating the action mechanisms of MWJD for treating gynecopathy of CCBS syndrome.</p><p><b>METHODS</b>Female SD rats were randomly divided into the normal group, the model group, and the treatment group. Rats in the model group and the treatment group were placed in the ice water (0 -1 degrees C) to establish the rat model of CCBS syndrome. Meanwhile, MWJD was given by gastrogavage to rats in the treatment group for 2 weeks. Rats were decapitated during the oestrous cycle. The serum levels of estradiol (E2), progestone (P), testosterone (T) were detected. The bilateral ovarian tissues were taken out and operated on ice. The total bilirubin (TBIL), superoxide dismutase (SOD), malondialdehyde (MDA), total antioxygen capacity (T-AOC) were detected after homogenate.</p><p><b>RESULTS</b>Compared with the normal group, serum levels of E2, P, T, and ovarian levels of TBIL, SOD, T-AOC all obviously decreased in the model group, and the ovarian MDA level increased, showing statistical difference (P <0. 05, P <0.01). After treated by MWJD, when compared with the model group, serum levels of E2, P, T, and ovarian levels of TBIL, SOD, T-AOC increased, the ovarian MDA level decreased, showing statistical difference (P <0.05, P <0.01). All indices except serum T level and the ovarian MDA content restored to the normal levels (all P>0.05).</p><p><b>CONCLUSIONS</b>The ovarian functions are abnormal in CCBS model rats. MWJD could restore the ovarian functions by improving the ovarian oxidative damage state, thus treating gynecopathy of CCBS syndrome.</p>

[Establishment of gynecological asthenia cold syndrome model].

OBJECTIVE: To establish the model of gynecological asthenia cold syndrome by simulating the etiology of gynecological asthenia cold syndrome depending on 'pathogenic cold impairing yang" in Chinese medicine theory. METHODS: The female SD rats were randomly divided into the model group and the normal group by randomized digital table, 20 in each group. Rats in the model group were placed in 0 degrees C - 1 degree C ice water, and the ice water was placed in 4 degrees C refrigerator, twice daily, 20 min each time, for a total of 30 days. The body temperature was determined and the changes of the estrous cycle were observed every day. When the body temperature decreased (with statistical difference from those of the normal group), and vaginal smears showed disordered estrous cycle, the model was successfully established. Rats were sacrificed during the diestrus period, the correlative indices including reproductive endocrine hormones in blood [follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol ( E2 ), testosterone (T), and progestone (P)], the thyroid function [triiodothyronine (T3), tetraiodothyronine (T4), thyrotropic-stimulating hormone (TSH)], the adrenal function [plasma adrenocorticotropic hormone (ACTH) and cortisol (Cor)], the cellular immune function [serum interleukin-2 (IL-2)], and energy tests [including plasma cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), the ratio of cAMP/cGMP, lactate dehydrogenase(LDH)], an the index of thymus, spleen, uterus, and ovary were detected. RESULTS: Compared with the normal group, rats in the model group appeared chill, dim-color hair, purple and dark ears and claw. They were depressed, scrunched, quiet, and clumsy. They liked to stay together. Their water intake and appetite were reduced, body weight lost, body temperature significantly lowered. They passed loose stool. Their estrous cycle and diestrus were prolonged. Their plasma cGMP content, cAMP/cGMP ratio, LDH, serum IL-2 content, E2, P, T, T3, LH, TSH were significantly lowered to some extent. Their thymus index and the ovary index significantly decreased, showing significant difference (P < 0.05). CONCLUSION: The rat model of gynecological asthenia cold syndrome prepared by extending the frozen time at refrigerator and ice water immersion was in line with clinical features of gynecological asthenia cold syndrome.

Binary competitive adsorption of naphthalene compounds onto an aminated hypercrosslinked macroporous polymeric adsorbent.

An aminated hypercrosslinked macroporous polymeric adsorbent was synthesized and characterized. Adsorption isotherms for 1-amino-2-naphthol-4-sulfonic acid (1,2,4-acid) and 2-naphthol obtained from various binary adsorption environments can be well fitted by Freundlich equation, which indicated a favorable adsorption process in the studied range. Adsorption for 1,2,4-acid was an endothermic process in comparison with that for 2-naphthol of an exothermic process. 2-naphthol molecules put a little influence on the adsorption capacity for 1,2,4-acid. However, the adsorption to 1,2,4-acid depressed that to 2-naphthol in a large extent for the stronger electrostatic interaction between 1,2,4-acid and adsorbent. The predominant mechanism can be contributed to the competition for adsorption sites. And the simultaneous environment was confirmed to be helpful to the selective adsorption towards 1,2,4-acid based on the larger selectivity index.