Experimental Demonstration of Efficient Harmonic Generation via Surface Plasma Compression with Lasers.

The efficiency of high-order harmonic generation from a relativistic laser interacting with solid targets depends greatly on surface plasma distribution. The usual method of enhancing efficiency involves tuning the plasma scale length carefully by improving the laser contrast. Here, we experimentally demonstrate that efficient harmonics can be achieved directly by compressing large-scale surface plasma via the radiation pressure of a circularly polarized normally incident prepulse. The harmonic generation efficiency obtained by this method is comparable to that obtained with optimized plasma scale length by high-contrast lasers. Our scheme does not rely on high-contrast lasers and is robust and easy to implement. Thus, it may pave a way for the development of intense extreme ultraviolet sources and future applications with high repetition rates.

[Clinical study of hysteroscopical electroresection of FIGO type 3 myoma].

Objective: To analyze the clinical application and to evaluate the efficiency of hysteroscopical electroresection of International Federation of Gynecology and Obstetrics (FIGO) type 3 myoma. Methods: The clinical data of patients who underwent hysteroscopical electroresection single FIGO type 3 myoma in Obstetrics and Gynecology Hospital of Fudan University from January 2019 to October 2021 were collected retrospectively. The clinical symptoms, myoma size, location, operation time, intraoperative bleeding, surgical complications and postoperative follow-up were recorded, and the subsequent pregnancy outcomes were followed-up. Results: Totally 35 patients with FIGO type 3 myoma were included in this study. The average age was (36.6±4.7) years old, the diameter of myoma was (4.0±1.2) cm (range: 2.0-5.8 cm). The rate of complete resection of myoma in one operation was 86% (30/35), the average operation time was (41±15) minutes (range: 20-65 minutes), and the average intraoperative bleeding was (24±18) ml (range: 5-150 ml). No complications such as uterine perforation, massive hemorrhage, hyperhyderation syndrome and infection occurred in all patients perioperation. There were 20 cases with significant increase of menstruation before operation, the cure rate and effective rate of hysteroscopical electroresection of FIGO type 3 myoma were 75% (15/20) and 95% (19/20). There were 24 patients with fertility requirements, their average follow-up time was (14.5±6.8) months, the pregnancy rate within 1 year after operation was 79% (19/24), and the average postoperative pregnancy time was (5.8±3.4) months. There were 15 cases who had completed delivery after operation, including 10 cases of vaginal delivery and 5 cases of cesarean section, and none of them had uterine rupture. Conclusions: Hysteroscopy could effectively resect FIGO type 3 myoma. Hysteroscopical electroresection of FIGO type 3 myoma is minimally invasive with rapid postoperative recovery and could achieve pregnancy in a short time, which is not only helpful to reduce the amount of menstruation, but also beneficial for the prognosis of fertility. It should be carried out by the experienced hysteroscopists.

[Clinical features of young inpatients with angle-closure glaucoma].

Objective: To analyze the composition of diagnosis in young inpatients with angle-closure glaucoma and to compare the clinical characteristics between primary angle-closure glaucoma (PACG) and secondary angle-closure glaucoma (SACG). Methods: This was a retrospective case series study. Angle-closure glaucoma patients aged 40 years or younger and hospitalized in Zhongshan Ophthalmic Center from January 2012 to December 2019 were included. The clinical diagnosis, gender, onset age, and results of general ophthalmic examination, A-scan ultrasonography measurements, ultrasound biomicroscopy, optical coherence tomography and visual field were recorded. The proportions and composition of PACG and SACG, as well as the misdiagnosis ratio, were analyzed. The onset age, visual acuity, visual field, and ocular parameters were compared between patients with PACG and SACG. Non-normally distributed data were represented by M (Q1, Q3). The difference between groups were compared by Mann-Whitney U test, Chi square test, and independent sample t test. Results: A total of 243 patients (243 eyes) were included in this study. The mean onset age was (28±9) years, and the male-to-female ratio was 1∶1.79. There were 93 patients (38.3%) in PACG group and 150 (61.7%) in SACG group. The age of PACG [34(28, 38)] was older than that of SACG [28(19, 34)], and the proportion of males in the SACG group (44.0%, 66/150) was significantly higher than that in the PACG group (22.6%, 21/93) (Z=-5.34, χ2=11.46; both P<0.01). Nanophthalmos (22.7%, 34 cases), autosomal recessive bestrophinopathy (ARB) (19.3%, 29 cases), uveitis (18.7%, 28 cases) and retinitis pigmentosa (14.0%, 21 cases) were the most common causes of SACG. The best corrected visual acuity (logarithm of the minimum angle of resolution) was 0.10 (0.00, 0.48) and 0.40 (0.06, 1.00), mean deviation of visual field was -8.07 (-27.49, -2.09) and -15.04 (-28.75, -5.97) dB, and subfoveal choroidal thickness was (452.3±130.7) and (396.3±120.9) µm in the PACG and SACG groups, respectively. The differences were statistically significant (Z=-4.86, -2.14; t=2.37; all P<0.05). There was no statistical difference in intraocular pressure, cup-to-disc ratio, extent of peripheral anterior synechia, central anterior chamber depth, lens thickness, mean retinal nerve fiber layer thickness between these two groups (all P>0.05). The misdiagnosis ratio was 10.7% (26/243) in all patients, while the misdiagnosis ratio of the SACG group (16.7%, 25/150) was higher than that of the PACG group (1.1%, 1/93) (χ²=14.61, P<0.001). Conclusions: PACG, nanophthalmos and ARB are the most common etiologies in young inpatients with angle-closure glaucoma. Compared to PACG, patients with SACG are younger, with worse visual acuity and more severe visual field defects, and easier to be misdiagnosed.

[Application of vaginoscopy in the diagnosis and treatment of occult vaginal high-grade squamous intraepithelial lesions].

Objective: To investigate the application value of vaginoscopy in the diagnosis and treatment of vaginal squamous intraepithelial lesions post hysterectomy. Methods: From May 2017 to June 2020, patients with high-risk (HR)-HPV infection and (or) cytological abnormalities after hysterectomy in Obstetrics and Gynecology Hospital Affiliated to Fudan University were examined by colposcopy, and those who were not satisfied with exposure under colposcopy were further examined by vaginoscopy. The role of vaginoscopy in the detection and treatment of occult vaginal squamous intraepithelial lesions was analyzed. Results: A total of 153 patients with HR-HPV infection and (or) cytological abnormalities, and inadequate colposcopy were enrolled in this study. The average age was (49.8±8.1) years. All cases were successfully performed vaginoscopy with no vaginal perforation, no bladder and intestinal injury. During vaginoscopy, 11 (7.2%, 11/153) cases with unclear high-grade squamous intraepithelial lesion (HSIL) boundary under colposcopy were found with clear HSIL boundary, and new HSIL was found in 23 (15.0%, 23/153) cases. According to colposcopy, there were 89 cases of normal or inflammation, 45 cases of vaginal low-grade squamous intraepithelial lesion (LSIL) and 19 cases of vaginal HSIL. According to vaginoscopy, there were 56 cases of normal or chronic inflammation, 55 cases of vaginal LSIL, 40 cases of vaginal HSIL (including 2 cases of vaginal HSIL could not exclude cancer) and 2 cases of vaginal carcinoma. There were significant difference between colposcopy and vaginoscopy (P<0.01). The missed diagnosis rate of vaginal apex blind curettage under colposcopy was 54.8%. For the 40 cases with vaginal HSIL under vaginoscopy, 15 cases were completely curettaged with vaginal endoscopic claw forceps, and 22 cases were vaporized by Versapoint. Among the 37 cases of vaginal HSIL treated by vaginoscopy, 34 cases were followed up for 6 months with 31 cases of vaginal HSIL cured. The cure rate was 91.2% (31/34), the positive rate of HR-HPV decreased from 100.0% (34/34) to 79.4% (27/34). Conclusion: For patients with inadequate exposure of vaginal apex during colposcopy posthysterectomy, vaginoscopy is helpful to detect the occult lesions in the vaginal apex, and it could also be used for the treatment of vaginal squamous intraepithelial lesions.

[Risk stratified management of cervical adenocarcinoma in situ based on cone margin state].

Objective: To investigate the hierarchical management scheme of cervical adenocarcinoma in situ (AIS) based on cervical conization margin state. Methods: All medical records of 249 patients diagnosed as AIS by loop electrosurgical excision procedure (LEEP) conization from Jan. 2010 to Dec. 2015 in Obstetrics and Gynecology Hospital of Fudan University were retrospectively reviewed, to explore the relationship between the status of the resection margin and the residual lesion after LEEP, and the multivariate logistic regression method was used to analyze the related factors that affect the residual lesion after LEEP in cervical AIS patients. Results: (1) The age of 249 cervical AIS patients was (40±8) years old (range: 23-71 years old). Of the 249 patients, 19 (7.6%, 19/249) had residual lesions; 69 cases were pathologically diagnosed as AIS after LEEP, and the residual lesion rate was 13.0% (9/69), which was significantly higher than that of AIS + high-grade squamous intraepithelial lesion [5.6% (10/180); χ2=3.968,P=0.046]; 33 cases were multifocal lesions, the residual rate of lesions was 21.2% (7/33), which was significantly higher than that of single focal lesions patients [5.6% (12/216); χ2=7.858, P=0.005]; 181 patients underwent endocervical curettage (ECC) before surgery, the residual rate of lesions in ECC-positive patients was 14.0% (14/100) , significantly higher than that of ECC-negative patients [4.9% (4/81); χ2=4.103, P=0.043]. (2) Among 249 cases of AIS patients, the positive rate of resection margins after LEEP was 35.3% (88/249); the residual rate of lesions in patients with positive resection margins (14.8%, 13/88) was significantly higher than those with negative margins [3.8%(6/156); χ2=9.355, P=0.002]. The age of patients underwent total hysterectomy after LEEP was (43±7) years old, which was significantly higher than that of patients who did not undergo total hysterectomy [(37±8) years old; t=6.518, P<0.01].Among the patients underwent total hysterectomy after LEEP, 3 cases (2.0%, 3/152) had fertility requirements, while 38 cases (39.2%, 38/97) did not underwent total hysterectomy, the difference between the two groups was statistically significant (χ2=59.579, P<0.01). Among the 152 patients who underwent total hysterectomy after LEEP, the residual rate of lesions was 11.8% (18/152); the residual rate of lesions in patients with positive resection margins was significantly higher than that of patients with negative resection margins [18.8% (12/64) vs 7.0% (6/86); χ2=4.861, P=0.028]. The median follow-up time of 97 patients who did not undergo total hysterectomy after LEEP was 32 months (range: 4-70 months). During the follow-up period, 3 cases of cervical AIS recurrence (3.1%, 3/97) and were followed by hysterectomy,no invasive adenocarcinoma were seen. (3) Multivariate logistic regression analysis showed that the positive resection margin (OR=4.098, 95%CI: 1.235-13.595, P=0.021), multifocal lesions (OR=5.464, 95%CI: 1.494-19.981, P=0.010) were independent risk factors that affected the residual lesions in patients with cervical AIS after LEEP. Conclusions: The cervical AIS patients after LEEP conization suggested be stratified by cone margin state as the first-line stratified index, age and fertility needs as the second-line stratified management index. The individualized management plan should be developed based on comprehensive assessment of high-risk factors of residual lesions.

[Value of high-resolution anoscopy in the diagnosis of anal precancerous lesions].

Objective: To evaluate the value of high-resolution anoscopy (HRA) in the diagnosis of anal precancerous lesions. Methods: A retrospective cohort study was performed, which included 142 patients who underwent HRA in vulvar and anal clinic at Obstetrics and Gynecology Hospital of Fudan University from January 2014 to December 2019. With the perianal and anal canal biopsy pathology as the "gold standard", the diagnostic value of HRA and specific findings for anal precancerous lesions were evaluated and the clinical characteristics of patients with anal precancerous lesions were analyzed. Results: (1) Agreement between HRA and anal pathology were 76.6% (95/124, perianal) and 70.0% (84/120, anal canal), in which there was no significant difference (χ2=1.365, P=0.243). The strength of agreement with weighted Kappa statistic were 0.604 (perianal) and 0.455 (anal canal) respectively. HRA diagnosis were more overestimated [16.9% (21/124) in perianal and 25.0% (30/120) in anal canal] than underestimated [6.5% (8/124) in perianal and 5.0% (6/120) in anal canal]. The sensitivity and specificity of HRA in the diagnosis of anal precancerous lesions were all more than 60.0% (perianal lesions: 97.8% and 74.7%; anal canal lesions: 90.9% and 66.7%), and Youden's index were >0.5 (perianal lesions: 0.725; anal canal lesions: 0.576). The positive predictive value of HRA in anal canal lesions (50.8%) was lower than that in perianal lesions (68.8%). (2) The thin acetowhite epithelium was the most common finding in anal precancerous lesions, and its performance in anal canal lesions (the sensitivity, the specificity and the Youden's index were 78.8%, 62.4% and 0.412, respectively) were better than those in perianal lesions (the sensitivity, the specificity and the Youden's index were 28.9%, 96.9% and 0.258, respectively). The dense acetowhite epithelium and vascular patterns were only seen in anal canal lesions. Lugol's iodine was little useful for perianal lesions (the incidence of iodine negative was 0),but quite useful to evaluate the canal lesions (the positive predictive value was 83.3% and the negative predictive value was 82.3%). (3) The average age of patients with low-grade anal precancerous lesion was (37±10) years old, and high-grade anal precancerous lesion was (42±11) years old, and there was significant difference between them (P=0.034). Age curve showed that the peak age was 30-40 years old. Vulvar intraepithelial lesion was the risk factor of anal precancerous lesions (χ2=4.284, P<0.05). Conclusions: HRA patterns are reliable in the diagnosis of anal precancerous lesions, which is important for guiding biopsy. However, it is easy to be overestimated, especially for anal canal lesions. The acetowhite epithelium is the most common finding in anal precancerous lesions, but vascular patterns are only seen in anal canal lesions.

WHITE STRIPE LEAF8, encoding a deoxyribonucleoside kinase, is involved in chloroplast development in rice.

KEY MESSAGE: WSL8 encoding a deoxyribonucleoside kinase (dNK) that catalyzes the first step in the salvage pathway of nucleotide synthesis plays an important role in early chloroplast development in rice. The chloroplast is an organelle that converts light energy into chemical energy; therefore, the normal differentiation and development of chloroplast are pivotal for plant survival. Deoxyribonucleoside kinases (dNKs) play an important role in the salvage pathway of nucleotides. However, the relationship between dNKs and chloroplast development remains elusive. Here, we identified a white stripe leaf 8 (wsl8) mutant that exhibited a white stripe leaf phenotype at seedling stage (before the four-leaf stage). The mutant showed a significantly lower chlorophyll content and defective chloroplast morphology, whereas higher reactive oxygen species than the wild type. As the leaf developed, the chlorotic mutant plants gradually turned green, accompanied by the restoration in chlorophyll accumulation and chloroplast ultrastructure. Map-based cloning revealed that WSL8 encodes a dNK on chromosome 5. Compared with the wild type, a C-to-G single base substitution occurred in the wsl8 mutant, which caused a missense mutation (Leu 349 Val) and significantly reduced dNK enzyme activity. A subcellular localization experiment showed the WSL8 protein was targeted in the chloroplast and its transcripts were expressed in various tissues, with more abundance in young leaves and nodes. Ribosome and RNA-sequencing analysis indicated that some components and genes related to ribosome biosynthesis were down-regulated in the mutant. An exogenous feeding experiment suggested that the WSL8 performed the enzymic activity of thymidine kinase, especially functioning in the salvage synthesis of thymidine monophosphate. Our results highlight that the salvage pathway mediated by the dNK is essential for early chloroplast development in rice.

[The 473rd case: renal failure, abdominal pain, and mental abnormality].

A 43-year-old male presented with elevated serum creatinine for 4 years and developed abdominal pain for 3 days. He started peritoneal dialysis 2 months ago. Dialysis-related peritonitis was ruled out and acute gastroenteritis was diagnosed. The patient was administrated with ertapenem 500 mg/d. An acute mental abnormality developed 3 days later. After excluded organic encephalopathy, ertapenem was discontinued for the suspicion of antibiotic-related encephalopathy. The frequency of peritoneal dialysis was increased to accelerate the clearance of antibiotics. However, the metal abnormality became even more severe. Then a diagnosis of Wernick-Korsakoff syndrome was considered. After the administration of high dose vitamin B(1), the mental disorder dramatically relieved. Vitamin B(1) 30 mg/d is maintained during peritoneal dialysis and the mental disorder does not relapse.

Evaluating the effects of mobile health intervention on weight management, glycemic control and pregnancy outcomes in patients with gestational diabetes mellitus.

PURPOSE: To explore the effects of mobile health (mHealth) intervention on pregnancy weight management, blood glucose control and pregnancy outcomes. METHODS: A total of 124 patients with gestational diabetes mellitus (GDM) were selected. Patients were randomly divided into two groups. The 60 patients in the control group received standard outpatient treatment, while the remaining 64 patients received a nurse's online guidance both through a mobile medical App installed on their phone and through regular offline clinical treatment in the mHealth group. Patients were treated for an average of 13 weeks and general conditions, compliance, blood glucose, glycosylated hemoglobin, weight gain, pregnancy, and neonatal outcomes were monitored in both groups longitudinally. RESULTS: The mHealth group demonstrated higher levels of compliance (83.3 ± 12.5% vs. 70.4 ± 10.1%, t = - 6.293, df = 122, p < 0.001), lower frequency of outpatient service (8.1 ± 1.3 vs. 11.2 ± 1.1, t = 14.285, df = 122, p < 0.001), lower hemoglobin A1C before delivery (4.7 ± 0.2 vs. 5.3 ± 0.3, t = 13.216, df = 122, p < 0.001) as well as the rates of off-target measurements both fasting (4.6 ± 0.4% vs. 8.3 ± 0.6%, t = 40.659, df = 122, p < 0.001) and 2 h post-prandial (7.9 ± 0.7% vs. 14.7 ± 0.8%, t = 50.746, df = 122, p < 0.001). Weight gain in the mHealth group was less than control group (3.2 ± 0.8 vs. 4.8 ± 0.7, t = 11.851, df = 122 p < 0.001). CONCLUSION: Mobile health intervention management of gestational diabetes mellitus improves patients' compliance and blood glucose control, and reduces weight gain, thereby reducing the rates of complications in both pregnant women and fetuses during delivery during pregnancy.

[Analysis of recurrence and its influencing factors in patients with cervical HSIL within 24 months after LEEP].

Objective: To analyze clinical outcome of high grade squamous intraepithelial lesion (HSIL) within 24 months after loop electrosurgical excision procedure (LEEP), and to explore risk factors of recurrent cervical HSIL, the risk of progress into cervical cancer and methods of follow-up. Methods: This retrospective study was carried out on 1 005 patients who underwent LEEP, diagnosing with HSIL after LEEP from January 2011 to December 2013 at Obstetrics and Gynecology Hospital Affiliated to Fudan University to confer the difference between non-recurrent group and recurrent group 24 months after the LEEP conization. Patients were followed with ThinPrep cytologic test (TCT), high risk HPV (HR-HPV) test, colposcopy guided biopsy. Results: A total of 1 005 cases were enrolled in this study with HSIL in the LEEP specimen, no residual HSIL in the 6-month follow up, and have follow up records in 24 months after LEEP. HSIL recurred in 5 cases, microinvasive carcinoma in 1 case, low grade squamous intraepithelial lesion (LSIL) in 17 cases at 12 months follow-up. HSIL recurred in 8 cases, LSIL in 11 cases, adenocarcinoma in situ in 1 case, and invasive cervical carcinoma in 1 case in Ⅰb1 stage at 24 months after LEEP. The recurrence rate was 1.3% (13/1 005) , and the progression rate was 0.3% (3/1 005) . There was no significant difference in age, length, circumference and width of LEEP between recurrent and non-recurrent patients (P>0.05) . The recurrence rate was highest in the endocervical positive margin group with 3/16, which was higher than ectocervical positive margin and negative margin (P<0.01, P=0.040, respectively). The recurrence rate of endocervical positive margin group and fibrous interstitial positive margin group showed no significant difference (P=0.320) . There was no significant difference between ectocervical positive margin and negative margin [2.8% (2/72) vs 0.7% (6/882), P=0.117]. Postoperative cytological examination combined with HR-HPV detection has a high sensitive and high negative predictor value of HSIL recurrence with both 100.0%. Multivariate logistic regression analysis showed that positive endocervical positive margin, abnormal follow-up cytological examination and positive HR-HPV after LEEP were independent factors affecting recurrence of HSIL patients after LEEP (P<0.05). Conclusions: Age, length, circumference and width of LEEP have no effect on recurrence within 24 months after HSIL. The high risk factors for HSIL recurrence within 24 months after LEEP in HSIL patients include: positive HPV, abnormal cytology, and positive endocervical positive margin. Colposcopy biopsy and endocervical curettage have important role in diagnosing HSIL recurrence and progression.

[Combined anluohuaxianwan and entecavir treatment significantly improve the improvement rate of liver fibrosis in patients with chronic hepatitis B virus infection].

Objective: To explore the improvement rate of liver fibrosis in patients with chronic hepatitis B virus infection who received entecavir alone or in combination with anluohuaxianwan for 78 weeks. Methods: Patients with chronic HBV infection were randomly treated with entecavir alone or in combination with anluohuaxian for 78 weeks. Ishak fibrosis score was used for blind interpretation of liver biopsy specimens. The improvement in liver fibrosis condition before and after the treatment was compared. Student's t test and non-parametric test (Mann-Whitney U-Test and Kruskal-Wallis test) were used to analyze the measurement data. The categorical variables were analyzed by Chi-square test method and Spearman's rank correlation coefficient was used to test bivariate associations. Results: Liver fibrosis improvement rate after 78 weeks of treatment was 36.53% (80/219) and the progression rate was 23.29% (51/219). The improvement of liver fibrosis was associated to the degree of baseline fibrosis and treatment methods (P < 0.05). The improvement rate of hepatic fibrosis in patients treated with anluohuaxianwan combined with entecavir at baseline F < 3 (54.74%, 52/95) was significantly higher than that in patients treated only with entecavir (33.33%, 16/48), P = 0.016 and the progression rate of hepatic fibrosis (13.68%, 13/95) was lower than that in patients treated alone (18.75%, 9/48), P = 0.466. In patients with baseline F < 3, the proportion of patients with improved and stable liver fibrosis in the combined treatment group (68.1%, 32/47) was higher than that in the treatment group alone (51.7%, 15/29). Conclusion: Combined anluohuaxianwan and entecavir treatment can significantly improve the improvement rate of liver fibrosis in patients with chronic hepatitis B virus infection. Furthermore, it has the tendency to improve the stability rate and reduce the rate of progression of liver fibrosis.

Energy enhancement of the target surface electron by using a 200 TW sub-picosecond laser.

One order of magnitude energy enhancement of the target surface electron beams with central energy at 11.5 MeV is achieved by using a 200 TW, 500 fs laser at an incident angle of 72° with a prepulse intensity ratio of 5×10-6. The experimental results demonstrate the scalability of the acceleration process to high electron energy with a longer (sub-picosecond) laser pulse duration and a higher laser energy (120 J). The total charge of the beam is 400±20 pC(E>2.7 MeV). Such a high orientation and mono-energetic electron jet would be a good method to solve the problem of the large beam divergence in fast ignition schemes and to increase the laser energy deposition on the target core.

Inhibition of arginase via jugular infusion of Nω-hydroxy-nor-l-arginine inhibits casein synthesis in lactating dairy cows.

A previous in vitro study revealed that Arg elicits positive effects on casein synthesis through alterations of the Arg-ornithine pathway in bovine mammary epithelial cells. The main purpose of this work was to determine the effects of arginase inhibition using Nω-hydroxy-nor-l-arginine (nor-NOHA) on milk protein synthesis in vivo. Six healthy Chinese Holstein cows with similar body weight (550.0 ± 20 kg; means ± standard deviation), parity (4), body condition score (3.0), milk yield (21.0 ± 1.0 kg), and days in milk (80 ± 2) were selected and randomly assigned to 3 treatments in a replicated 3 × 3 Latin square design with 22 d for each period (7 d for infusion and 15 d for washout). The treatments were (1) control: saline infusion; (2) nor-NOHA: infusion of 125 mg/L of nor-NOHA; (3) nor-NOHA + Arg: infusion of 125 mg/L of nor-NOHA with 9.42 g/L of Arg. The activity of enzymes related to Arg metabolism, milk protein synthesis, and expression of AA transporters was determined. The infusion of nor-NOHA decreased the activity of arginase but had no effect on the activity of ornithine decarboxylase and nitric oxide synthase in serum, and these responses were the same at the gene expression level in mammary gland. In addition, the infusion of nor-NOHA also reduced protein and fat synthesis in milk but had no effect on milk yield. When Arg was infused with nor-NOHA, the activity of total arginase, ornithine decarboxylase, and nitric oxide synthase, and the concentration of casein, protein, and fat in milk did not change compared with the nor-NOHA group, but the milk protein yield, the expression of some Arg transporters (SLC7A5 and SLC7A8), and milk yield increased. Overall, results verified previous in vitro findings indicating that synthesis of casein protein is closely regulated by the Arg-ornithine pathway in bovine mammary gland.

[Influence of celastrol on toll-like receptor 4-mediated signaling pathway in the free fatty acids-induced HepG2 cells].

Objective: To investigate the effect and mechanism of celastrol on free fatty acids (FFAs)-induced HepG2 cells. Methods: Cultured human HepG2 cells were transfected with toll-like receptor 4 (TLR4) siRNA, and the interference efficiencies were examined by real-time PCR. HepG2 cells were treated with FFAs and celastrol, and the untreated cells were used as a normal control (NC). Deposition of lipids in the HepG2 cells were visualized by Oil Red O staining. The protein expression of TLR4 and downstream inflammatory mediators [myeloid differentiation factor 88 (MyD88), nuclear factor (NF)-κBp65, interleukin (IL)-1ß and tumor necrosis factor α (TNF-α)] in the HepG2 cells were determined by Western blotting. The significance of the data obtained was evaluated using analysis of variance (ANOVA). Results: Red lipid droplets were extensively deposited in HepG2 cells after 0.5 mmol/L FFAs induction and significantly decreased in the celastrol-treated group. The protein expression of TLR4 and downstream inflammatory mediators (MyD88, NF-κBp65, IL-1ß and TNF-α) in the FFAs-induced HepG2 cells increased significantly compared with those of the NC group (all P<0.05), and were suppressed in TLR4 siRNA-treated and celastrol-treated group (TLR4: 0.69±0.14, 1.63±0.12 vs 2.46±0.23; MyD88: 1.21±0.12, 1.35±0.18 vs 1.62±0.19; NF-κBp65: 1.69±0.14, 1.54±0.36 vs 2.19±0.47; IL-1ß: 1.51±0.16, 1.45±0.38 vs 1.82±0.27; TNF-α: 1.60±0.14, 1.41±0.29 vs 1.88±0.19) (all P<0.01). Co-treatment with TLR4 siRNA and celastrol further reduced the expression of inflammation mediators compared with those of the TLR4 siRNA-treated group (MyD88: 1.09±0.23 vs 1.21±0.12; NF-κBp65: 1.24±0.20 vs 1.69±0.14; IL-1ß: 1.28±0.31 vs 1.51±0.16; TNF-α: 1.10±0.29 vs 1.60±0.14) (all P<0.01). Conclusion: Celastrol exerts its protective effect partly via inhibiting the TLR4-mediated signaling pathways in the steatotic HepG2 cells.

[The protective effect of Xuebijing on paraquat-induced HK-2 cells apoptosis and the underlying mechanisms].

Objective: To investigate the protective effect and mechanism of Xuebijing (XBJ) on paraquat (PQ) -induced apoptosis in Human kidney cell line-2 (HK-2) cells. Methods: Routinely cultured HK-2 cells, (1) Cell growth inhibition experiment after PQ and XBJ intervention: PQ was divided into 0、200、400、800、1600 and 3200 µmol/L PQ groups, and the cell survival rate was detected after intervening 24、48 and 72 h. XBJ was divided into 0、5、10、20、40 mg/ml XBJ groups, and the cell survival rate was detected after intervening 24、48 and 72 h.To determine the rational drug concentration and the duration of action of XBJ and PQ. (2) PQ-induced HK-2 cell growth inhibition experiment antagonized by XBJ: The cells were divided into normal control group, PQ group (800 µmol/L) and PQ+XBJ group (The cells were pretreated with 5、10 and 20 mg/ml XBJ for 1 h, then cultured with PQ of 800 µmol/L) , After cultured 24 h、48 h and 72 h separately, the cell survival rate was detected. (3) HK-2 cells were divided into normal control group、PQ group (800 µmol/L PQ cultured for 24 h) 、PQ+XBJ group (pretreated with 10 mg/ml XBJ for 1 h, and then 800 µmol/L PQ cultured for 24 h) and XBJ group (10 mg/ml XBJ cultured 24 h). The apoptosis of cells was detected by flow cytometry. The protein expression of Bcl-2 and BAX in each group was detected by Western blotting. The expressions of caspase-3 and caspase-9 were detected by caspase-3 and caspase-9 activity kit active. Results: (1) PQ could significantly reduced the survival rate of HK-2 cells and showed time and concentration dependence. The survival rate of HK-2 cells was about 55% after 800 µmol/L PQ contacted 24 h, XBJ under 20 mg/ml was no significant effect on the survival rate of HK-2 cells after cultured 72 h. (2) Compared with the PQ group, the survival rate of HK-2 cells of PQ+XBJ group was significantly increased (P<0.05). (3) Compared with the normal control group, the cell apoptosis rate of PQ group was significantly increased (P<0.05). Compared with the PQ group, the cell apoptosis rate of PQ+XBJ group was significantly decreased (P<0.05). (4) Compared with the normal control group, Bcl-2 protein expression in PQ group was significantly decreased and BAX protein expression in PQ group was significantly increased (P<0.05) ; compared with PQ group, Bcl-2 protein expression in PQ+XBJ group was significantly increased, BAX protein expression in PQ+XBJ group was significantly decreased (P<0.05). (5) Compared with the normal control group, the activities of caspase-3 and caspase-9 in PQ group were significantly increased (P<0.05). Compared with PQ group, the activities of caspase-3 and caspase-9 in PQ+XBJ group were decreased significantly (P<0.05) . Conclusion: XBJ (10 mg/ml) has obvious protective effect on HK-2 cell injuried by PQ (800 µmol/L) , It can improve the survival rate of cells through reducing the apoptosis of HK-2 cells which induced by PQ.

The symptom phenotype of oncology outpatients remains relatively stable from prior to through 1 week following chemotherapy.

Some oncology outpatients experience a higher number of and more severe symptoms during chemotherapy (CTX). However, little is known about whether this high risk phenotype persists over time. Latent transition analysis (LTA) was used to examine the probability that patients remained in the same symptom class when assessed prior to the administration of and following their next dose of CTX. For the patients whose class membership remained consistent, differences in demographic and clinical characteristics, and quality of life (QOL) were evaluated. The Memorial Symptom Assessment Scale (MSAS) was used to evaluate symptom burden. LTA was used to identify subgroups of patients with distinct symptom experiences based on the occurrence of the MSAS symptoms. Of the 906 patients evaluated, 83.9% were classified in the same symptom occurrence class at both assessments. Of these 760 patients, 25.0% were classified as Low-Low, 44.1% as Moderate-Moderate and 30.9% as High-High. Compared to the Low-Low class, the other two classes were younger, more likely to be women and to report child care responsibilities, and had a lower functional status and a higher comorbidity scores. The two higher classes reported lower QOL scores. The use of LTA could assist clinicians to identify higher risk patients and initiate more aggressive interventions.

Short communication: Arginase inhibition reduces the synthesis of casein in bovine mammary epithelial cells.

The main purpose of this work was to determine the effect of arginase inhibition via Nω-hydroxy-nor-l-arginine (nor-NOHA) on casein synthesis in bovine mammary epithelial cells (BMEC). Passage 2 BMEC isolated from dairy cows were seeded to 6-well plates and randomly divided into 4 treatments: (1) control [Dulbecco's modified Eagle medium:Nutrient Mixture F-12 medium (DMEM/F12)]; (2) nor-NOHA (DMEM/F12 + 1 mmol/L nor-NOHA); (3) nor-NOHA + arginine (DMEM/F12 + 1 mmol/L nor-NOHA + 3.2 mmol/L Arg); and (4) nor-NOHA + ornithine (DMEM/F12+ 1 mmol/L nor-NOHA + 1 mmol/L Orn). Then, we determined the activity of enzymes related to Arg metabolism and casein synthesis in BMEC and the proliferation of cells. The addition of nor-NOHA reduced the activity of arginase and ornithine decarboxylase but had no effect on the activity of nitric oxide synthase, and these responses were the same at the gene expression level. In addition, supplementation of nor-NOHA in BMEC reduced cellular proliferation and casein synthesis. Addition of Arg to nor-NOHA resulted in cellular proliferation and casein synthesis similar to that of nor-NOHA alone. In contrast, addition of Orn to the medium with nor-NOHA increased the synthesis of casein and cellular proliferation compared with Nor-NOHA. In conclusion, suppression of the Arg-arginase-Orn pathway reduced casein synthesis and cellular proliferation, which indicated that this pathway is an important regulator of the synthesis of casein in BMEC.

Effects of dietary physically effective neutral detergent fiber content on the feeding behavior, digestibility, and growth of 8- to 10-month-old Holstein replacement heifers.

The objective of this study was to evaluate the effects of dietary physically effective neutral detergent fiber (peNDF) content on the feeding behavior, digestion, ruminal fermentation parameters, and growth of 8- to 10-mo-old dairy heifers and to predict the adequacy of dietary fiber in growing dairy heifers. Twenty-four Holstein dairy heifers (245 ± 10.8 d of age, 305.6 ± 8.5 kg initial live weight) were randomly divided into 4 treatments with 6 replicates as a completely randomized design. During the 60-d period with a 10-d adaptation, heifers were offered 1 of 4 diets, which were chemically identical but included different peNDF8.0 (particle size is >8 mm and <19 mm) content (% DM): 10.8, 13.5, 18.0, or 19.8%, which was achieved by chopping forage into different lengths (fine = 1 cm, short = 3 cm, medium = 5 cm, and long = 7 cm). The concentrate and silage were mixed and fed restrictedly and exclusive of forage (Chinese ryegrass hay) were offered ad libitum. The body weight and frame size of the heifers were measured every 15 d during the experimental period. Samples of the rumen content (2 h after the morning feeding) were taken for pH, ammonia, and volatile fatty acid determination. The dry matter intake and average daily gain of the heifers were not significantly affected by peNDF8.0 content. The body frame size (including withers height, body length, and heart girth) of the heifers was not increased significantly by enhanced peNDF8.0 content. Ruminal pH and ammonia concentration were both increased with increasing dietary peNDF8.0 content. The ruminal total volatile fatty acid concentration and percentage of acetate and butyrate profiles were not significantly affected by dietary peNDF8.0 content. However, the enhanced peNDF8.0 content led to a decrease in the propionate percentage. The ratio of acetate to propionate in the 13.5% treatment was highest among the treatments. Increasing the particle size and dietary peNDF8.0 content resulted in increased eating and chewing time but had no effect on rumination time. Heifer total eating and chewing time and eating and chewing time per kilogram of dry matter intake were increased with increasing dietary peNDF8.0 content. The apparent digestibility of acid detergent fiber and crude protein was improved with an increasing content of dietary peNDF8.0. The results suggest that an optimal or advisable dietary particle size and peNDF8.0 content improves chewing activity, rumen fluid pH, and ruminal fermentation. The data based on feeding behavioral and growth responses of heifers as well as rumen fermentation and digestion by improving total eating and chewing time indicate that 18.0% dietary peNDF8.0 content is the most suitable for 8- to 10-mo-old Holstein heifers.

Establishment of a cell line with high transfection efficiency from zebrafish Danio rerio embryos and its susceptibility to fish viruses.

A cell line ZBE3 isolated from a continuous cell culture derived from zebrafish Danio rerio blastomeres by clonal growth was characterized. ZBE3 cells had been subcultured for >120 passages since the initial primary culture of the blastomeres. The ZBE3 cells grow stably at temperature from 20 to 32° C with an optimum temperature of 28° C in ESM2 or ESM4 medium with 15% foetal bovine serum (FBS). The optimum FBS concentration for ZBE3 cell growth ranged from 15 to 20%. Cytogenetical analysis indicated that the modal chromosome number of ZBE3 cells was 50, the same as the diploid chromosome number of D. rerio. Significant cytopathic effect was observed in ZBE3 cells after infection with redspotted grouper nervous necrosis virus, Singapore grouper iridovirus and grass carp reovirus, and the viral replication in the cells was confirmed by real-time quantitative PCR and transmission electron microscopy, indicating the susceptibility of ZBE3 cells to the three fish viruses. After transfected with pEGFP-N3 plasmid, ZBE3 cells showed a transfection efficiency of about 40% which was indicated by the percentage of cells expressing green fluorescence protein. The stable growth, susceptibility to fish viruses as well as high transfection efficiency make ZBE3 cells be a useful tool in transgenic manipulation, fish virus-host cell interaction and immune response in fish.