Objective: To explore the effects of different drying methods on the quality of medicinal materials, and screen out the optimal drying process of Centella asiatica. Methods: The whole fresh grass of C. asiatica were dried by hot air, drying in the sun, drying in the sun and hot air combined, drying in the shade, microwave and vacuum respectively. Meanwhile, the drying time and rate were determined. The characters, identification, inspection, and leachable content of C. asiatica were analyzed by the method of pharmacopoeia. The content of asiaticoside, madecassoside, asiatic acid, madecassic acid, kaempferol-3-O-tutinoside, kaempferol, and quercetin were detected by HPLC analysis; The weighted scoring method was used to sort the comprehensive evaluation of multiple indexes. Results: Different drying methods consume different time, among which drying in the sun, shade and drying at 50 ℃ for more than 100 h, and the average drying rate was 24.83%. The effects of different drying methods on the properties of medicinal materials are mainly reflected in color and odor, among which 50-70 ℃ hot air drying had a better color, which was light green, and the odor of hot air drying and microwave drying at 80 ℃ and 85 ℃ also changed significantly. Although there were some differences in moisture and ash content, both of them met the pharmacopoeia standards. The drying method also had certain effects on the leachable, the maximum was 45.70%, and the minimum content of dry extract was 29.67%. The highest content of the total active ingredient was determined by HPLC using the method of drying in the shade, which was 83.032 mg/g, and the lowest was is 75 ℃ hot air drying, which was 40.982 mg/g. The highest total content of madecassoside and asiaticoside was 80 ℃ hot air drying, and the lowest was 75 ℃ hot air drying. Weighted score in the top three of line was 70 ℃, dried at 50 ℃ after drying in the sun, hot air drying at 50 ℃, and 85 ℃ hot air drying ranked the bottom. Conclusion: In summary, the suitable drying method for the production area of C. asiatica was 70 ℃ hot air drying.

OBJECTIVE:To explore basic technology for synthesis of active ingredients of Ophiocordyceps xuefengensis,and provide necessary technical support for comprehensive development of O. xuefengensis sourse. METHODS:Submerged fermenta-tion method was used to cultivate the mycelium,achieving efficient synthesis of active ingredients by controlling medium composi-tion and cultivation conditions. Using the bacteria as starting strain,the effects of different carbon sources (sucrose,glucose and soluble starch),different nitrogen sources (peptone,yeast extract powder,yeast extract,sodium nitrate,potassium nitrate and urea),different vitamin B(vitamin B1 and vitamin B complex)and different initial pH(pH was set at 4,5,6,7,8 and 9,re-spectively)on mycelial growth,extracellular and intracellular polysaccharide synthesis,cordycepin synthesis and intracellular triter-penoid synthesis were investigated to screen the optimal medium composition. RESULTS:The optimal carbon source,nitrogen source,vitamin B and initial pH were sucrose,yeast extract powder,vitamin B1 and 8,respectively. High biomass and metabolite accumulation levels can be obtained when carbon source was sucrose,nitrogen source was yeast extract powder,adding 0.1 g/L vi-tamin B1 with initial pH of 8. CONCLUSIONS:O. xuefengensis can efficiently accumulate metabolites,and achieve the optimiza-tion of strain cell growth and synthesis of active metabolite by optimizing and controlling the fermentation process.

Objective To investigate and compare the different effects of soluble adult wornl antigen(SWA)and soluble egg antigen(SEA)of Schistosoma japonicum on the apoptosis and cell-cycle of routine CD4~+T cells.Methods Purified CD4~+T ceUs from normal C57BL/6 mice were cultured with CFSE labeled antigen presenting clls in the presence of different stimuli for 36 h.Flow cytometry(FCM)was used to detect the apoptosis of CD4~+T cells by fluorescence conjugated caspase-3 antibodie staining.The flow cytometry was used to analyze the cell-cycle of CD4~+T cells cultured as described above for 96 h by propidium iodide staining.Results Compared with the apoptosis percentage of CD4~+T cells[(1.24±0.29)%]in the SEA stimulated group,that in the SWA stimulated group[(1.52±0.38)%]did not show statistically significant difference(P>0.05).Compared with the cell percentages in G1 phase[(78.91±2.98)%],S phase[(7.39±0.85)%]and G2/M phase[(10.69±1.05)%] in the SWA stimulated group,that of the G1 phase[(59.42±1.32)%]was significantly lower,but those in the S phase[(21.07±O.88)%] and G2/M phase[(18.88±1.21)%]were significantly increased in the SEA stimulated group(P<0.01).Conclusions There is no statistically significant difference between the apoptosis levels of CD4~+T ceHs stimulated by SWA and SEA.However,SEA significantly promotes the progression of the cell-cycle of CD4~+T cells compared with SWA.

Spermatogenesis, a continuous course of cell proliferation and differentiation, depends on reproductive hormones. FSH and LH-induced T are the main hormones regulating spermatogensis. Intratesticular T is one of the key factors in maintaining spermatogenesis, while FSH is just as important for origination and maintenance of normal spermatogenesis. Sertoli cells are the pivot of hormonal regulation. Interestingly, these reproductive hormones also regulate sperm cell apoptosis spermatogenesis. Further studies on the hormonal regulation of spermatogenesis provide a base for the development of safe and recoverable contraceptives for males.
Androgens ; physiology ; Apoptosis ; Contraception ; Follicle Stimulating Hormone ; physiology ; Humans ; Luteinizing Hormone ; physiology ; Male ; Spermatogenesis

Objective: To observe the efficacy of long-retaining scalp acupuncture plus interactive training in improving upper- extremity dysfunction in cerebral stroke patients. Methods: Ninety-five patients with upper-extremity dysfunction after cerebral stroke were randomized into two groups, with 48 cases in the treatment group and 47 cases in the control group. Conventional internal medicine treatment was offered to both groups. In both groups, Anterior Oblique Line of Vertex-temporal (MS 6, the middle 2/5) and Posterior Oblique Line of Vertex-temporal (MS 7, the middle 2/5) were selected from the same side of the brain lesion (the side apposing to the hemiplegic limb) for scalp acupuncture treatment. In the treatment group, the scalp acupuncture needles were retained for 7 h, in combination with interactive training, while the needles were also retained for 7 h in the control group but without interactive training. Prior to treatment and at 2-week and 4-week treatment, the two groups were scored using the functional test for the hemiplegic upper extremity-Hong Kong (FTHUE-HK) and simplified Fugl-Meyer assessment-upper extremity (FMA-UE). Results: The total effective rate was 97.9％ in the treatment group, higher than 74.5％ in the control group (P<0.01). The FTHUE-HK score was higher at 2-week and 4-week treatment than before treatment in both groups, presenting statistically significant intra-group differences (all P<0.001); the FTHUE-HK score was higher at 4-week treatment than at 2-week treatment in both groups, presenting statistically significant intra-group differences (both P<0.001). At 2-week and 4-week treatment, the FTHUE-HK score was higher in the treatment group than in the control group, showing significant between-group differences (both P<0.05). During the whole treatment process, the treatment group had higher FTHUE-HK scores compared with the control group, but there was no statistical significance comparing the change of the score between the two groups at 2-week treatment (P>0.05), while the between-group difference in the change of the score was statistically significant at 4-week treatment (P<0.05). The FMA-UE score was higher at 2-week and 4-weeks treatment than before treatment in both groups, presenting statistically significant intra-group differences (all P<0.001); the FMA-UE score was higher at 4-week treatment than at 2-week treatment in both groups, presenting statistically significant intra-group differences (both P<0.001). At 2-week and 4-week treatment, the FMA-UE was higher in the treatment group than in the control group, and the between-group differences were statistically significant (both P<0.01). The FMA-UE score rose gradually with the increase of treatment session, and there was statistical significance comparing the change of the score between the two groups at 2-week and 4-week treatment, respectively (both P<0.05). Conclusion: Long-retaining scalp acupuncture plus interactive training results in more significant efficacy than long-retaining scalp acupuncture alone in improving the upper-limb dysfunction after cerebral stroke and the advantage becomes more notable after 2-week consecutive treatment.

Objective:To investigate the effects of adherent-invasive Escherichia coli ( E. coli) LF82 on the structure and function of intestinal barrier in mice with ulcerative colitis (UC). Methods:Twenty-four specific pathogen free (SPF) C57BL/6 mice were divided into UC with E. coli LF82 group, UC group and healthy control group with eight mice in each group. The UC mice model was induced by dextran sulfate sodium (DSS). One week before modeling, the mice of UC with E. coli LF82 group were intragastric administrated with 1×10 9 colony-forming unit (CFU) E. coli LF82 to colonize the bacteria strain. The effects of E. coli LF82 on colitis of mice with UC were evaluated by disease activity index (DAI), gross morphological injury score, colonic mucosal injury index (CMDI), myeoloperoxidase (MPO) activity and pathological features. The ultrastructure and the changes of cytoskeleton F-actin of mice colonic tissues were detected by transmission electron microscope (TEM) and direct immunofluorescence. The ability of colonic mucin production and degree of fibrosis were estimated by periodic acid Schiff reaction (PAS) stain and sirius red stain. T test, least significant difference, repeated measurement analysis of variance and one-way analysis of variance were used for statistical analysis. Results:On the fourth, fifth, sixth and seventh day after the modeling, the DAI scores of UC with E. coli LF82 group were all higher than those of UC group ((2.53±0.38) points vs. (2.01±0.53) points, (3.02±0.62) points vs. (2.67±0.24) points, (3.13±0.61) points vs. (2.20±0.24) points, (3.27±0.28) points vs. (2.20±0.69) points, respectively), and the differences were statistically significant ( t=3.37, 2.25, 9.56 and 10.24, all P<0.05). The gross morphological injury score of mice colon of UC with E. coli LF82 group was higher than that of UC group ((6.17±1.94) points vs. (2.83±0.98) points), and the difference was statistically significant ( t=-3.75, P<0.05). The CMDI and MPO activity of UC with E. coli LF82 group were both higher than those of UC group ((16.80±2.79) points vs. (11.80±3.11) points, (729.3±77.5) U/mg vs. (594.4±31.9) U/mg), and the differences were statistically significant ( t=-2.83; mean difference=134.82, 95% confidence interval ( CI) 72.12 to 197.51; both P<0.05). The results of TEM showed that the E. coli LF82 could invade the submucosa of colon and caused further injury of colonic tissues in mice. The distribution of cytoskeleton F-actin of mice colonic tissues changed. The results of PAS staining showed that the percentages of PAS positive cells of UC with E. coli LF82 group and UC group were both lower than that of healthy control group ((32.40±8.02)% and (41.90±8.99)% vs. (57.70±11.52)%), and the difference was statistically significant ( F=17.63, P<0.01). The percentage of PAS positive cells of UC with E. coli LF82 group was lower than that of UC group, and the difference was statistically significant (mean difference=-9.50, 95% CI -18.33 to -0.67, P<0.05). The results of sirius red staining showed that the villous epithelium of colon mucosa of UC with E. coli LF82 group was partially injured and collagen fibers hyperplasia was serious. The area ratios of collagen fiber of UC with E. coli LF82 group and UC group were both higher than that of healthy control group ((51.83±5.78)% and (37.11±5.59)% vs. (15.41±2.25) %), and the difference was statistically significant ( F=86.72, P<0.01). The area ratio of collagen fiber of UC with E. coli LF82 group was higher than that of UC group, the difference was statistically significant (mean difference=14.83, 95% CI 8.91 to 20.76, P<0.05). Conclusions:E. coli LF82 can aggravate DSS-induced colitis in UC mice, leading to changes in colon ultrastructure and cytoskeleton, it can also reduce the ability of mucus secretion of colon of mice and increase the degree of colonic tissues fibrosis.

Objective To evaluate surgical treatment of chronic tibial osteomyelitis of Cierny-Mader type Ⅳ with Ilizarov technique and lesion osteotomy. Methods From January 2010 to May 2016, 39 patients with chronic tibial osteomyelitis of Cierny-Mader type Ⅳ were treated at our center. They were 33 males and 6 females, 8 to 54 years of age (average, 33.8 years). After debridement and lesion osteotomy, the tibia was fixated with Ilizarov external fixator. Bone was transported to the bone defect after corticotomy was performed on the proximal and/or distal tibial metaphyses simultaneously. Bifocal corticotomy was per-formed in 11 cases, proximal corticotomy in 21 cases, and distal corticotomy in 7 cases. The transport began 3 to 5 days after operation at a speed of 0.5 to 1.0 mm/d initially. The speed was lowered according to the bone healing and pain. Radiographic examination was done every 2 weeks to observe transporting deviation and osteogenesis in the transporting area. The transporting was adjusted whenever any abnormality was observed. The bone transporting lasted for 50 to 130 days (average, 62.4 days). Results The patients were fol-lowed up for 11 to 49 months (average, 21 months). All the soft tissue wounds healed uneventfully and there was no relapse of osteomyelitis. The bone defects in the 32 cases were reconstructed primarily. Nonunion of fracture ends happened in 5 cases and nonunion of the bone lengthening zone in 2 cases. The 7 cases of nonunion were healed after secondary bone grafting. Malalignment happened in 5 cases, 4 of which responded to timely adjustment of the external fixation and one of which had to receive secondary bone grafting after failure in adjustment of the external fixation. Ankle joint dysfunction occurred in 7 cases, 5 of which re-sponded to functional exercise and 2 of which accepted joint dysfunction because they refused surgery after unsatisfactory functional exercise. Pin tract infection of different severities occurred in 9 cases, one of which was treated by replacement of the K-wires under local anesthesia and the other 8 of which responded to rein-forced dressing change. Conclusions Chronic tibial osteomyelitis of Cierny-Mader typeⅣcan be treated by Ilizarov technique and lesion osteotomy. However, the Ilizarov technique should be improved because of the risks of multiple complications which can be reduced significantly by strengthening postoperative instruction, nursing, and regular follow-up.

Adult ; Cell Cycle Proteins/genetics* ; Codon, Nonsense/genetics* ; Consanguinity ; Humans ; Infertility, Male/genetics* ; Male ; Oligospermia/genetics* ; Pedigree ; Spermatogenesis/genetics*

OBJECTIVES: To investigate the clinical treatment outcomes and the changes of the outcomes over time in extremely preterm twins in Guangdong Province, China. METHODS: A retrospective analysis was performed for 269 pairs of extremely preterm twins with a gestational age of <28 weeks who were admitted to the department of neonatology in 26 grade A tertiary hospitals in Guangdong Province from January 2008 to December 2017. According to the admission time, they were divided into two groups: 2008-2012 and 2013-2017. Besides, each pair of twins was divided into the heavier infant and the lighter infant subgroups according to birth weight. The perinatal data of mothers and hospitalization data of neonates were collected. The survival rate of twins and the incidence rate of complications were compared between the 2008-2012 and 2013-2017 groups. RESULTS: Compared with the 2008-2012 group, the 2013-2017 group (both the heavier infant and lighter infant subgroups) had lower incidence rates of severe asphyxia and smaller head circumference at birth (P<0.05). The mortality rates of both of the twins, the heavier infant of the twins, and the lighter infant of the twins were lower in the 2013-2017 group compared with the 2008-2012 group (P<0.05). Compared with the 2008-2012 group, the 2013-2017 group (both the heavier infant and lighter infant subgroups) had lower incidence rates of pulmonary hemorrhage, patent ductus arteriosus (PDA), periventricular-intraventricular hemorrhage (P-IVH), and neonatal respiratory distress syndrome (NRDS) and a higher incidence rate of bronchopulmonary dysplasia (P<0.05). CONCLUSIONS There is a significant increase in the survival rate over time in extremely preterm twins with a gestational age of <28 weeks in the 26 grade A tertiary hospitals in Guangdong Province. The incidences of severe asphyxia, pulmonary hemorrhage, PDA, P-IVH, and NRDS decrease in both the heavier and lighter infants of the twins, but the incidence of bronchopulmonary dysplasia increases. With the improvement of diagnosis and treatment, the multidisciplinary collaboration between different fields of fetal medicine including prenatal diagnosis, obstetrics, and neonatology is needed in the future to jointly develop management strategies for twin pregnancy.
Bronchopulmonary Dysplasia/epidemiology* ; Female ; Gestational Age ; Humans ; Infant ; Infant, Extremely Premature ; Infant, Newborn ; Pregnancy ; Respiratory Distress Syndrome, Newborn/epidemiology* ; Retrospective Studies ; Treatment Outcome