The aim of this investigation was to determine the optimal storage medium for testicular hypothermic transportation and identify the ideal concentration for the application of the protective agent 5-aminolevulinic acid (5-ALA). Furthermore, this study aimed to explore the underlying mechanism of the protective effects of 5-ALA. First, we collected and stored mouse testicular fragments in different media, including Hank's balanced salt solution (HBSS; n = 5), Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F12; n = 5), and alpha-minimum essential medium (αMEM; n = 5). Storage of testicular tissue in HBSS preserved the integrity of testicular morphology better than that in the DMEM/F12 group ( P < 0.05) and the αMEM group ( P < 0.01). Testicular fragments were subsequently placed in HBSS with various concentrations of 5-ALA (0 [control], 1 mmol l -1 , 2 mmol l -1 , and 5 mmol l -1 ) to determine the most effective concentration of 5-ALA. The 2 mmol l -1 5-ALA group ( n = 3) presented the highest positive rate of spermatogonial stem cells compared with those in the control, 1 mmol l -1 , and 5 mmol l -1 5-ALA groups. Finally, the tissue fragments were preserved in HBSS with control ( n = 3) and 2 mmol l -1 5-ALA ( n = 3) under low-temperature conditions. A comparative analysis was performed against fresh testes ( n = 3) to elucidate the underlying mechanism of 5-ALA. Gene set enrichment analysis (GSEA) for WikiPathways revealed that the p38 mitogen-activated protein kinase (MAPK) signaling pathway was downregulated in the 2 mmol l -1 5-ALA group compared with that in the control group (normalized enrichment score [NES] = -1.57, false discovery rate [FDR] = 0.229, and P = 0.019). In conclusion, these data suggest that using 2 mmol l -1 5-ALA in HBSS effectively protected the viability of spermatogonial stem cells upon hypothermic transportation.
Male ; Animals ; Testis/cytology* ; Aminolevulinic Acid/pharmacology* ; Mice ; Organ Preservation/methods* ; Organ Preservation Solutions/pharmacology* ; Cryopreservation/methods*

OBJECTIVE: To explore the effect of Lactobacillus reuteri on testicular injury in mice exposed to neonicotinoid insecticides (NNI). METHODS: Fifteen C57BL/6 male mice were randomly divided into control group (CTRL group), exposure group (NNI group) and Lactobacillus intervention group (NNI-L group). The mice in CTRL group were given 0.02ml/g of 0.5% carboxymethyl cellulose sodium solution by gavage for 14 days. The mice in NNI group were given 0.02 ml/g of NNI mixture by gavage for 14 days. The mice in NNI-L group were given 0.02 ml/g of NNI mixture by gavage and 5×108cfu/ml of Lactobacillus reuteri powder solution for 14 days. Then, the histomorphology and function of testicle were evaluated by hematoxylin-eosin staining, immunofluorescence staining and RNA sequencing. RESULTS: Compared with CTRL group, the thickness of testicular seminiferous epithelium in the NNI group was significantly thinner. And the decline in the number of spermatogenic cells and sperm was observed. And the expression of spermatogonial stem cell marker UCHL1 was down-regulated which was significantly improved in NNI-L group compared with the NNI group. The abnormal expressions of hormone and sperm methylation related genes in testis of NNI group were detected by RNA sequencing, with significant down-regulation being found in NPFF and IGF2. While the expression of HSD3B8 was significantly up-regulated. The abnormal expression of these genes could be significantly improved after oral administration of Lactobacillus reuteri. CONCLUSION Testicular spermatogenesis and endocrine function can be damaged by NNI exposure. And oral administration of Lactobacillus reuteri protects testis from the adverse effects of NNI toxicity.
Animals ; Male ; Limosilactobacillus reuteri ; Testis/pathology* ; Mice ; Mice, Inbred C57BL ; Insecticides/toxicity* ; Neonicotinoids/toxicity* ; Probiotics ; Spermatogenesis/drug effects*

The present study explored the effect of co-amorphous technology in improving the dissolution rate and stability of silybin based on the puerarin-silybin co-amorphous system prepared by the spray-drying method. Solid-state characterization was carried out by powder X-ray diffraction(PXRD), polarizing microscopy(PLM), Fourier transform infrared spectroscopy(FT-IR), differential scanning calorimetry(DSC), etc. Saturated powder dissolution, intrinsic dissolution rate, moisture absorption, and stability were further investigated. The results showed that puerarin and silybin formed a co-amorphous system at a single glass transition temperature which was higher than that of any crude drug. The intrinsic dissolution rate and supersaturated powder dissolution of silybin in the co-amorphous system were higher than those of the crude drug and amorphous system. The co-amorphous system kept stable for as long as three months under the condition of 40 ℃, 75% relative humidity, which was longer than that of the single amorphous silybin. Therefore, the co-amorphous technology could significantly improve the dissolution and stability of silybin.
Calorimetry, Differential Scanning ; Desiccation ; Drug Compounding/methods* ; Drug Stability ; Silybin ; Solubility ; Spectroscopy, Fourier Transform Infrared ; Technology ; X-Ray Diffraction

Objective: To compare the short-term and long-term outcomes between robotic-assisted and laparoscopic-assisted radical right hemicolectomy in patients with adenocarcinoma of the right colon. Methods: Retrospective review of a prospectively collected database identified 288 right colon cancer patients who underwent either robotic-assisted (n=57) or laparoscopic-assisted right hemicolectomy (n=231) between October 2014 and October 2020 at Department of Gastrointestinal Surgery, the Affiliated Hospital of Qingdao University. There were 161 males and 127 females, aging (60.3±12.8) years (range: 17 to 86 years). After propensity score matching as 1∶4 between robotic-assisted and laparoscopic-assisted right hemicolectomy, there were 56 cases in robotic group and 176 cases in laparoscipic group. Perioperative outcomes and overall survival were compared between the two groups using t test, Wilcoxon rank sum test, χ² test, Fisher exact test, Kaplan-Meier method and Log-rank test, respectively. Results: The total operative time was similar between the robotic and laparoscopic group ((206.9±60.7) minutes vs. (219.9±56.3) minutes, t=-1.477, P=0.141). Intraoperative bleeding was less in the robotic group (50 (20) ml vs. 50 (50) ml, Z=-4.591, P<0.01), while the number of lymph nodes retrieved was significantly higher (36.0±10.0 vs. 29.0±10.1, t=4.491, P<0.01). Patients in robotic group experienced significantly shorter hospital stay, shorter time to first flatus, and defecation (t: -2.888, -2.946, -2.328, all P<0.05). Moreover, the overall peri-operative complication rate was similar between robotic and laparoscopic group (17.9% vs. 22.7%, χ²=0.596,P=0.465). The 3-year overall survival were 92.9% and 87.9% respectively and the 3-year disease-free survival rates were 83.1% and 82.6% with no statistical significance between the robotic and laparoscopic group (P>0.05). Conclusions: Compared to laparoscopic-assisted right hemicolectomy, robot-assisted right hemicolectomy could improve some short-term clinical outcomes. The two procedures are both achieving comparable survival.
Colectomy ; Colonic Neoplasms/surgery* ; Female ; Humans ; Laparoscopy ; Male ; Prognosis ; Propensity Score ; Retrospective Studies ; Robotic Surgical Procedures ; Treatment Outcome

Objective: Dianjixueteng is a geoherb in Yunnan Province, the source plant of which is Kadsura interior. However, the formation of this geoherb is not clear in genetic mechanism, in which genome size is the first step that should be known on the genomic level. In this study we aimed to estimate the genome sizes of source plants of K. interior and three related herbs K. heteroclita, K. longipedunculata, and K. coccinea by flow cytometry (FCM) and make a comparison. Methods: The genome sizes of K. interior, K. heteroclita, K. longipedunculata and K. coccinea, i.e., the source plants of Dianjixueteng and its relative medicinal materials, were estimated by FCM. The nuclei of K. interior were isolated using modified LB01 buffer, for the rest species, by the Galbraith's buffer. Results: The genome sizes of K. interior, K. heteroclita, K. longipedunculata, and K. coccinea were 7.36, 7.12, 7.01, and 5.15 pg/1C, respectively. Genome size of K. interior had no significant variation with those of K. heteroclita and K. longipedunculata (P = 0.296), which was significantly larger than that of K. coccinea. Conclusion: Genome size can not distinguish K. interior from K. heteroclita and K. longipedunculata, but could distinguish them from K. coccinea, which lays the foundation for future studies on genetic mechanism of the geoherb formation.

Objective: To evaluate the nutritional status by the Controlling Nutritional Status (CONUT) score and its association with the long-term prognosis in patients with acute heart failure (AHF). Methods: This prospective monocentric study consecutively enrolled patients admitted to our hospital for AHF from April 2012 to May 2016. Patients were divided into 3 groups based on the CONUT score at admission: normal (0-1), mild malnutrition (2-4) and moderate-severe malnutrition (5-12) groups. Baseline information was obtained and recorded within 24 hours after admission. All patients were followed up every 3 months by outpatient visit or telephone call until March 2019. The primary endpoint was all-cause mortality. The Kaplan-Meier survival curves and log-rank test were used to compare all-cause mortality between groups. Variables showing statistical significance in the univariate analysis were incorporated into multivariate Cox regression model to analyze the independent risk factors for all-cause mortality after discharge. Results: A total of 396 patients were enrolled in this study, including 114 patients with normal nutritional status, 200 patients with mild malnutrition and 82 patients with moderate-severe malnutrition. One hundred and fifty-eight patients died during a median follow-up of 34 (18, 46) months. The mortality was 32.4% (37/114), 39% (78/200) and 52.4% (43/82) in normal, mild malnutrition and moderate-severe malnutrition groups, respectively. The mortality was significantly higher in the moderate-severe malnutrition group than in normal nutrition group (P<0.05). However, there was no significant difference in mortality between normal and mild malnutrition group as well as between mild and moderate-severe malnutrition group (both P>0.05). Kaplan-Meier curves indicated that patients with high CONUT score group was at higher risk of all-cause mortality compared with those with low CONUT score (P=0.002). Cox proportional hazard analyses showed that the risk of all-cause mortality of moderate-severe malnutrition group was significantly higher than that of normal nutrition group (HR =1.648, 95%CI 1.021-2.660, P=0.041). Conclusions: The CONUT score of patients with AHF at admission is associated with the long-term prognosis. High CONUT score is an independent risk factor for all-cause mortality in AHF patients after discharge.
Heart Failure ; Humans ; Nutrition Assessment ; Nutritional Status ; Prognosis ; Prospective Studies ; Retrospective Studies

Anti-HIV Agents/therapeutic use* ; China ; Consensus ; HIV ; HIV Infections/prevention & control* ; Humans ; Pre-Exposure Prophylaxis

In order to improve the supersaturation and maintenance time of drug dispersion in curcumin self-nanoemulsion(CUR-SNEDDS), precipitation inhibitors(PPIs) were introduced to prepare curcumin supersaturated self-emulsion(CUR-SSNEDDS). The composition of CUR-SNEDDS prescriptions was selected through the solubility test, the compatibility of oil phase and surfactant, the investigation of the emulsifying ability of the surfactant and the drawing of the pseudo-ternary phase diagram. Analytic hierarchy process was used in combination with central composite design-response surface method to optimize the prescription. The type and dosage of precipitation inhibitors(PPIs) were selected to maintain the supersaturated concentration and duration of CUR in artificial gastrointestinal fluids. At the same time, polarizing microscope was used to evaluate the crystallization inhibition effect and the quality and in vitro release behavior of CUR-SSNEDDS. The prepared CUR-SSNEDDS prescription was capryol 90-kolliphor RH40-transcutol HP-Soluplus(7.93∶66.71∶25.36∶5), with the drug loading of(65.12±1.25) mg·g~(-1). CUR-SSNEDDS was transparent yellow, and the nanoemulsion droplets were spherical with uniform distribution. The emulsification time was(21.02±0.13) s, the average particle size was(57.03±0.35) nm, the polydispersity index(PDI) was(0.23 ± 0.01), and the Zeta potential was(-18.10±1.30) mV. CUR-SSNEDDS significantly inhibited the generation and growth of crystals after in vitro dilution. The supersaturation could be maintained above 10 within 2 h, and the dissolution rate and degree of CUR in artificial gastrointestinal fluid were significantly increased. Soluplus could effectively maintain the supersaturated state of CUR and enhance CUR dissolution in vitro.
Biological Availability ; Curcumin ; Emulsions ; Nanoparticles ; Particle Size ; Solubility ; Surface-Active Agents

Objective To investigate the clinical efficacy of heat-sensitive point medicinal moxibustion plus percutaneous administration of tetrandrine in treating lumbar intervertebral disc herniation. Methods One hundred and twenty patients with lumbar intervertebral disc herniation were randomized to treatment and control groups, 60 cases each. The treatment group received heat-sensitive point medicinal moxibustion plus percutaneous administration of tetrandrine and the control group, heat-sensitive point medicinal moxibustion alone. The VAS score and the JOA Score for Back Pain score were recorded in the two groups before and after treatment. The clinical therapeutic effects were compared between the two groups. Results There were statistically significant pre-/post-treatment differences in the JOA Score for Back Pain score and the VAS score in the two groups (P<0.01). There were statistically significant post-treatment differences in the JOA Score for Back Pain score and the VAS score between the treatment and control groups (P<0.01). The excellent and good rate was 80.0％ in the treatment group and 55.0％ in the control group; there was a statistically significant difference between the two groups (P<0.01). Conclusion Heat-sensitive point medicinal moxibustion plus percutaneous administration of tetrandrine is an effective way to treat lumbar intervertebral disc herniation.

OBJECTIVETo investigate the factors upgrading the International Society of Urological Pathology (ISUP) Gleason score using the specimens from preoperative prostatic biopsy and radical prostatectomy.

METHODSA total of 164 patients diagnosed with prostate cancer by biopsy underwent radical prostatectomy. We retrospectively analyzed their age, prostate volume, preoperative PSA level, PSA density (PSAD) , the time interval between biopsy and surgery, the number of positive punctures, positive surgical margin, seminal vesicle invasion, lymphatic invasion, and Gleason scores from biopsy and prostatectomy. We also determined the predictors of Gleason score upgrading by logistic regression analysis.

RESULTSOf the 164 cases analyzed, 95 (57.93% ) showed a consistency between the Gleason score of preoperative prostatic biopsy and that after radical prostatectomy, 55 (33.54% ) increased and 14 (8.52%) decreased after prostatectomy as compared with preoperative biopsy. The prostate volume (P < 0.01) and biopsy score (P < 0.05) were independent predictors of Gleason score upgrading. The risk of Gleason score upgrading was 27 times higher in the patients with the prostate volume ≤ 25 ml and 9 times higher in the 25-40 ml group than in the > 60 ml group (P < 0.05).

CONCLUSIONLow Gleason score of biopsy (≤ 6) and small prostate volume (≤ 40 ml) may be the predictors of Gleason score upgrading after radical prostatectomy.

Biopsy ; Humans ; Male ; Neoplasm Grading ; Organ Size ; Prostate-Specific Antigen ; blood ; Prostatectomy ; Prostatic Neoplasms ; classification ; surgery ; Retrospective Studies ; Risk Factors