Phylogeny and species diversity of the genus Helvella with emphasis on eighteen new species from China.

Helvella is a widespread, frequently encountered fungal group appearing in forests, but the species diversity and molecular phylogeny of Helvella in China remains incompletely understood. In this work, we performed comprehensive phylogenetic analyses using multilocus sequence data. Six datasets were employed, including a five-locus concatenated dataset (ITS, nrLSU, tef1-α, rpb2, hsp), a two-locus concatenated dataset (ITS, nrLSU), and four single-locus datasets (ITS) that were divided based on the four different phylogenetic clades of Helvella recognized in this study. A total of I 946 sequences were used, of which 713 were newly generated, including 170 sequences of ITS, 174 sequences of nrLSU, 131 sequences of tef1-α, 107 sequences of rpb2 and 131 sequences of hsp. The phylogeny based on the five-locus concatenated dataset revealed that Helvellas. str. is monophyletic and four phylogenetic clades are clearly recognized, i.e., Acetabulum clade, Crispa clade, Elastica clade, and Lacunosa clade. A total of 24 lineages or subclades were recognized, II of which were new, the remaining 13 corresponding with previous studies. Chinese Helvella species are distributed in 22 lineages across four clades. Phylogenetic analyses based on the two-locus concatenated dataset and four single-locus datasets confirmed the presence of at least 93 phylogenetic species in China. Among them, 58 are identified as known species, including a species with a newly designated lectotype and epitype, 18 are newly described in this paper, and the remaining 17 taxa are putatively new to science but remain unnamed due to the paucity or absence of ascomatal materials. In addition, the Helvella species previously recorded in China are discussed. A list of 76 confirmed species, including newly proposed species, is provided. The occurrence of H. crispa and H. elastica are not confirmed although both are commonly recorded in China. Citation: Mao N, Xu YY, Zhang YX, Zhou H, Huang XB, Hou CL, Fan L (2023). Phylogeny and species diversity of the genus Helvella with emphasis on eighteen new species from China. Fungal Systematics and Evolution 12: 111-152. doi: 10.3114/fuse.2023.12.08.

[Preliminary study on the etiological characteristics and clinical treatment of medication-related osteonecrosis of the jaws].

Objective: To summarize the etiological characteristics and clinical treatment effects of 17 patients with medication-related osteonecrosis of the jaw (MRONJ). Methods: The clinical data of 17 patients with MRONJ admitted to the Department of Oral and Maxillofacial Surgery, the First People's Hospital of Jinzhong, Shanxi Province, from July 2016 to December 2019 were retrospectively analyzed, including 9 males and 8 females, aged (63.6±9.6) years old (43-82 years old). Descriptive analysis of the primary disease, onset factors, site of disease, clinical manifestations, treatment methods, and treatment effects was conducted through follow-up for at least 1 year. Results: Among the primary diseases of the 17 cases, 12 were malignant tumors, and 5 were osteoporosis. There were 13 cases with a history of a trigger event (tooth extraction or unsuited removable denture). Six cases occurred in the maxilla, 10 cases occurred in the mandible, and 1 case involved both the upper and lower jaws. For the most common medication used, bisphosphonate was used in 16 cases including 5 cases with concomitant use of angiogenesis-inhibiting drugs. There was 1 case resulted from receptor activator of NF-κB ligand (RANKL) monoclonal antibody application. The duration of medication application was (10.1±3.9) months (3-18 months). All 17 cases were treated surgically. Totally 15 patients healed well after surgical treatment and the other 2 patients, who had poor soft tissue healing after surgery, healed well after a second operation. Conclusions: Tooth extraction might be a major trigger factor for the onset of MRONJ in the mandible. The disease was more possibly occured in the mandible than in the maxilla. Appropriate surgical treatment could achieve a good clinical outcome.

[AMG-102 inhibits proliferation and induces apoptosis of laryngeal squamous cell carcinoma cells by regulating c-Met/PI3K/Akt pathway].

Objective: To investigate the effects of c-Met inhibitor AMG-102 on the proliferation and apoptosis of laryngeal squamous carcinoma Hep-2 cells and the underlying mechanism. Methods: Laryngeal squamous carcinoma cell line Hep-2 cells were treated with 2.5, 5 and 10 µmol/L AMG-102, respectively. The proliferation activities of Hep-2 cells were detected by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide (MTT). The apoptotic rate of Hep-2 cells was detected by flow cytometry analysis and Hoechst staining. The mRNA expression levels of apoptosis-related genes were detected by real-time quantitative polymerase Chain reaction (RT-qPCR), and the protein expressions of c-Met/PI3K/AKT pathway were detected by western blot. Results: Compared with the control group, the proliferation rates of Hep-2 cells treated with 2.5, 5 and 10 µmol/L AMG-102 for 24 hours were (89.8±1.1)%, (79.8±1.0)% and (69.1±1.2)%, respectively; for 48 hours were (76.8±2.0)%, (60.2±1.1)% and (49.8±1.2)%, respectively; for 72 hours were (50.1±2.0)%, (41.5±1.1)% and (33.6±1.0), respectively, with significant differences (all P<0.05). The apoptotic rates of Hep-2 cells treated with 2.5, 5 and 10 µmol/L AMG-102 for 48 hours were (16.09±1.53)%, (27.51±2.02)% and (36.57±1.42)%, respectively, which were significantly higher than (3.62±0.10) % in the control group (all P<0.05). After treated with 2.5, 5 and 10 µmol/L AMG-102 for 48 hours, the relative expression levels of Bcl-2 mRNA in Hep-2 cells were 0.58±0.13, 0.38±0.12 and 0.20±0.13, respectively; the relative protein expression of p-Met were 80.0±3.8, 50.6±4.2 and 28.5±1.3, respectively; the relative protein expression of p-PI3K were 87.1±0.9, 54.2±1.2 and 21.0±1.2, respectively; the relative protein expression of p-AKT were 98.7±5.6, 56.9±3.2 and 32.2±4.3, respectively; which were significantly lower than those in the control group (all P<0.05). The relative expression levels of Bax mRNA were 1.78±0.13, 2.37±0.14 and 3.05±0.13, respectively, and the relative expression levels of caspase-3 mRNA were 1.98±0.14, 2.47±0.14 and 3.15±0.13, respectively, which were significantly higher than those in the control group (all P<0.05). Conclusion: c-Met inhibitor AMG-102 could inhibit the proliferation and induce apoptosis of laryngeal squamous carcinoma Hep-2 cells by regulating the c-Met/PI3K/Akt pathway.

[Effect of c-Met inhibitor AMG-102 on radiosensitivity in laryngeal squamous carcinoma cells].

Objective: To investigate the effect of c-Met inhibitor AMG-102 on proliferation and radiosensitivity in laryngeal squamous carcinoma cells. Methods: The effects of AMG-102 on proliferation and radiosensitivity of laryngeal squamous carcinoma cell lines Hep-2 and KBV200 were detected by 3-(4, 5-dimethy-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) assay and colony formation assay, respectively. The apoptosis of Hep-2 and KBV200 cells was detected by flow cytometry. The expression levels of c-Met, phospho-Met (p-Met), cleaved caspase-3 and Akt/p-Akt, Erk/p-Erk were detected by Western blot. Specific small interfering RNA targeting c-Met or plasmid of c-Met were transfected into Hep-2 and KBV200 cells to investigate the cell sensitivity to AMG-102. Results: Compared with KBV200 cells, Hep-2 cells were more sensitive to AMG-102 with IC(50) of 14 and 9 µmol/L, respectively. The relative expression levels of c-Met and p-Met proteins in Hep-2 cells were 194.48±0.57 and 177.76±1.53, respectively, which were significantly higher than those in KBV200 cells (171.24±1.00 and 115.37±0.56, respectively, P<0.001 for both). Exogenous hepatocyte growth factor (HGF) was added to increase the expression level of p-Met protein in KBV200 cells. The results showed that AMG-102 significantly reduced the expression of p-Met in KBV200 cells treated with HGF (P<0.001). Compared with the dimethyl sulfoxide (DMSO) group, AMG-102 treatment combined with radiotherapy significantly increased the radiosensitivity of Hep-2 cells (SER=1.28, P<0.001). However, AMG-102 had little effect on the radiosensitivity of KBV200 cells (SER=1.18, P=0.002). Compared with the 4 Gy radiotherapy alone group and the 5 µmol/L of AMG-102 alone treatment group, the apoptosis rate of Hep-2 cells in the combined treatment group was significantly increased. Meanwhile, the expression level of cleaved caspase-3 protein was also markedly increased. However, there were no significant changes in the apoptotic rate and cleaved caspase-3 expression in each treatment group of KBV200 cells. Compared with DMSO treatment group, the expression levels of p-Met, p-Akt and p-Erk were significantly decreased in the 4 Gy radiotherapy group, 5 µmol/L of AMG-102 treatment group and combined treatment group of Hep-2 cells. And the levels of p-Met, p-Akt and p-Erk in the combined treatment group were significantly lower than those in the 4 Gy radiotherapy alone group and 5 µmol/L of AMG-102 treatment alone group. By contrast, in KBV200 cells, the expression of p-Met, p-Akt and p-Erk in each group was not changed. The relative expression of p-Met in Hep-2 cells before and after radiotherapy at 30 min, 1 h, 4 h, 8 h, 24 h were 99.89±0.61, 138.62±1.00, 163.07±5.00, 87.80±1.85, 90.67±0.65 and 94.09±1.41, respectively. The level of p-Met was slightly increased after radiotherapy at 30 min and 1 h (P<0.001 for all), whereas it was significantly decreased from 4 h to 24 h after radiotherapy (P<0.05 for all). By contrast, the expression of p-Met in KBV200 cells did not change with time after radiotherapy (P>0.05). The sensitivity of Hep-2 cells to AMG-102 was decreased after silencing of c-Met, while the sensitivity of KBV200 cells to AMG-102 was not significantly changed (P>0.05). Moreover, the radiosensitivity of Hep-2 cells in c-Met knockdown group had a slightly increasing trend (SER=1.07, P=0.068). After the treatment with 10 µmol/L of AMG-102, the proliferation rate of c-Met ectopically expressed KBV200 cells was 60.05%±3.23%, It was significantly lower than that of the blank control 90.08%±1.04% and siRNA negative control (90.12%±1.01%, P<0.001). The results suggested that the overexpression of c-Met in KBV200 cells increased the radiosensitivity to AMG-102, whereas depletion of c-Met resulted in resistance to AMG-102 in Hep-2 cells. Furthermore, the radiosensitivity of KBV200 cells that overexpressed c-Met showed a decreased trend (SER=0.7, P=0.005). Conclusions: c-Met inhibitor AMG-102 has a significant inhibitory effect on the proliferation of c-Met overexpressing laryngeal squamous carcinoma cells, leading to increased radiosensitivity. It suggests that molecular targeted therapy against c-Met receptor is more effective in c-Met overexpressed subtype of laryngeal squamous cell carcinoma.

Prevention of Escherichia coli infection in broiler chickens with Lactobacillus plantarum B1.

Two studies were performed to assess the efficacy of Lactobacillus plantarum B1 in prevention of pathogenic Escherichia coli K88 gastrointestinal infection in broilers. In an in vitro study, L. plantarum B1 showed resistance to acid and bile and inhibited the growth of E. coli K88. Additionally, L. plantarum B1 exhibited high ability to adhere to broiler embryo ileal epithelium. In an animal trial, 240 broilers at 1 d of age were randomly assigned to one of 4 treatment arms: negative control (NC) broilers fed a basal diet and not challenged; positive control (PC) broilers fed a basal diet and challenged with E. coli K88; L. plantarum (LP) treatment broilers fed a basal diet containing 2 × 109 cfu/kg L. plantarum B1 and challenged with E. coli K88; and antibiotic treatment (Anti) broilers fed a basal diet supplemented with colistin sulfate (20 mg/kg) and challenged with E. coli K88. Broilers fed L. plantarum B1 had greater (P ≤ 0.05) BW than those in the PC treatment on d 14 and 28. Dietary L. plantarum B1 decreased (P < 0.05) E. coli counts in the cecal contents on d 10 and 14, and increased (P < 0.05) cecal lactic acid bacteria (LAB) on d 8, 10, 14, and 28 compared with the PC treatment. Dietary supplementation of L. plantarum B1 increased (P < 0.05) the ileal mucosal secretory IgA concentration and reduced (P < 0.05) IL-2, IL-4, IFN-γ, and tumor necrosis factor-α levels in the ileum. Overall, these results suggest dietary supplementation of L. plantarum B1 promotes growth performance, lowers cecal E. coli counts, and increases the population of cecal LAB, as well as improves intestinal mucosal immunity in E. coli K88-challenged broilers.

Fungal endophytes of South China blueberry (Vaccinium dunalianum var. urophyllum).

A total of 374 fungal endophyte strains were isolated from of Vaccinium dunalianum var. urophyllum (Ericaceae), a well-known cultivated blueberry in southern China. These fungal endophytes could be categorized into 25 morphotypes according to culture characteristics and molecular identification based on the internal transcribed spacer region. All of these isolates belonged to Ascomycota. Jaccard's (Jc) and Sorenson's similarity indices indicated that the species communities from the fruits and branches were closer to each other than to those from leaves. The leaves appeared to host the highest fungal biodiversity, and the fruits displayed the lowest diversity. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is the first on endophytic fungi isolated from fruits, branches and leaves of blueberry plants. The results contribute to the body of knowledge on the biocontrol of pathogens associated with blueberry and develop the improvement of plant growth. By comparing with the different fungal communities, the leaves appeared to host the highest biodiversity.

Effects of dietary Lactobacillus plantarum B1 on growth performance, intestinal microbiota, and short chain fatty acid profiles in broiler chickens.

Two experiments were conducted to determine the effects of Lactobacillus plantarum B1 on broiler performance, cecal bacteria, and ileal and cecal short chain fatty acids (SCFA). The study also determined whether it was necessary to feed Lactobacillus throughout the entire growth period or if the beneficial effects could be obtained by supplementation during the starter or finisher period only. Experiment 1 was conducted with 72 broilers assigned to 2 treatments (N=6). One treatment was the basal diet (Con), and the other was the basal diet supplemented with 2×10(9) cfu/kg L. plantarum B1 (Wh). In experiment 2, 144 one-day-old broilers were assigned to 4 treatments (N=6) including a basal diet (Con), the basal diet supplemented with 2×10(9) cfu/kgL. plantarum B1 during d one to 21 only (St), the basal diet supplemented with L. plantarum B1 during d 22 to 42 only (Fn), and, finally, the basal diet supplemented with L. plantarum B1 from d one to 42 (Wh). Experiment 1 showed that L. plantarum B1 enhanced broiler average daily gain (ADG) and feed conversion ratio (FCR). In experiment 2, during the starter period, broilers in the Wh and St treatments had higher ADG (P<0.05) than broilers in the Con and Fn, while during the finisher period, broilers in the Wh and Fn had higher ADG (P<0.01) and improved FCR (P<0.01) compared with broilers in the Con and St. On d 42, broilers in the Wh and Fn had decreased E. coli (P<0.05) and increased lactic acid bacteria (P<0.05) in their cecal digesta. L. plantarum B1 also increased (P<0.05) ileal mucosal sIgA as well as ileal and cecal SCFA. However, L. plantarum B1 had no effect on intestinal morphology. In conclusion,L. plantarum B1 plays a positive role in broilers. Supplementation during the finisher period or the entire growth period is superior to supplementation during the starter period only.

The antimicrobial peptide sublancin ameliorates necrotic enteritis induced by Clostridium perfringens in broilers.

Sublancin is an antimicrobial peptide produced by 168 containing 37 amino acids. The objective of this study was to investigate its inhibitory efficacy against both in vitro and in vivo. In the in vitro study, we determined that sublancin had a minimum inhibitory concentration of 8 µM against , which was much higher than the antibiotic lincomycin (0.281 µM). Scanning electron microscopy showed that sublancin damaged the morphology of . The in vivo study was conducted on broilers for a 28-d period using a completely randomized design. A total of 252 chickens at 1 d of age were randomly assigned to 1 of 6 treatments including an uninfected control; an infected control; 3 infected groups supplemented with sublancin at 2.88, 5.76, or 11.52 mg activity/L of water; and an infected group supplemented with lincomycin at 75 mg activity/L of water (positive control). Necrotic enteritis was induced in the broilers by oral inoculation of on d 15 through 21. Thereafter, the sublancin or lincomycin were administered fresh daily for a period of 7 days. The challenge resulted in a significant decrease in ADG ( < 0.05) and a remarkable deterioration in G:F ( < 0.05) during d 15 to 21 of the experiment. There was a sharp increase of numbers in the cecum ( < 0.05). The addition of sublancin or lincomycin reduced caecal counts ( < 0.05). The counts had a tendency to decrease in the lincomycin treatment ( = 0.051) but were the highest in the sublancin treatment (5.76 mg activity/L of water). A higher villus height to crypt depth ratio in the duodenum and jejunum as well as a higher villus height in the duodenum were observed in broilers treated with sublancin or lincomycin ( < 0.05) compared with infected control broilers. It was observed that sublancin and lincomycin decreased IL-1ß, IL-6, and tumor necrosis factor-α levels ( < 0.05) in the ileum compared with the infected control. In conclusion, although sublancin's minimum inhibitory concentration is much higher than lincomycin in vitro, less sublancin is needed to control necrotic enteritis induced by in vivo than lincomycin. These novel findings indicate that sublancin could be used as a potential antimicrobial agent to control necrotic enteritis.

Clozapine prescription and quality of life in Chinese patients with schizophrenia treated in primary care.

OBJECTIVE: Clozapine is frequently used to treat schizophrenia in China. Maintenance treatment for clinically stable patients with schizophrenia is usually provided by Chinese primary care physicians, but no study has investigated the frequency of its use prescribed by primary care physicians. This study described the frequency, demographic and clinical characteristics of clozapine treatment and its impact on insight and quality of life (QOL) of patients with schizophrenia treated in primary care in China. METHOD: A total of 623 patients with schizophrenia treated in 22 primary care services in Guangzhou, China in 2013 formed the study sample. Patients' socio-demographic and clinical characteristics including psychopathology, medication side effects and QOL were recorded using a standardized protocol and data collection. RESULTS: The frequency of clozapine prescription was 35.6% with a mean daily dose of 127.7±88.2 mg. There were no significant differences between the patients with and without clozapine in either of the QOL domains after controlling the confounding factors. Multiple logistic regression analyses revealed that patients on clozapine had younger age of onset, more hospitalizations, more severe extrapyramidal side effects, but better insight and fewer prescriptions of first generation antipsychotics. CONCLUSIONS: Clozapine use was found to be common and associated with better insight in patients with schizophrenia treated in primary care in China. Further examination of the rationale and appropriateness of clozapine in primary care in China is warranted.

Fermentation conditions influence the fatty acid composition of the membranes of Lactobacillus reuteri I5007 and its survival following freeze-drying.

Lactobacillus reuteri I5007 has well-documented adhesion properties and health benefits. Future industrial use of Lact. reuteri I5007 will require the development of effective fermentation procedures and high bacterial survival following drying. Therefore, this study was conducted to determine the impact of altering fermentation pH and temperature on the fatty acid composition of the bacterial membranes and subsequent survival of Lact. reuteri I5007 following freeze-drying. Initially, a response surface methodology was used to determine the optimal fermentation pH (5·7) and temperature (37°C), with regard to producing the maximum number of Lact. reuteti I5007 cells. However, when subjected to the optimal fermentation pH and temperature (control treatment), the subsequent survival of Lact. reuteri I5007 following freeze-drying was only 12·95%. Growth at a higher temperature (47°C) or at a neutral pH (pH 6·7) significantly increased the survival of Lact. reuteri I5007 following freeze-drying compared with the control. In contrast, an acidic pH (pH 4·7), or cold (27°C) and extremely cold (4°C) temperatures during fermentation significantly reduced Lact. reuteri I5007 survival following freeze-drying. The fatty acid composition of the membranes of Lact. reuteri I5007 was altered by the different fermentation conditions tested. An increase in the ratio of unsaturated fatty acids (UFA) to saturated fatty acids (SFA) in the bacterial membrane was associated with higher survival of Lact. reuteri I5007. In conclusion, it appears that the use of a higher temperature (47°C) or neutral pH (6·7) during fermentation resulted in increased survival of Lact. reuteri I5007 following freeze-drying. Significance and impact of the study: In this study, we found that a higher fermentation temperature or a neutral pH, rather than cold or acidic conditions, leads to increased survival of Lact. reuteri I5007 during subsequent freeze-drying. This finding has important implications for the future industrial production of this probiotic strain.

Superoxide dismutase recombinant Lactobacillus fermentum ameliorates intestinal oxidative stress through inhibiting NF-κB activation in a trinitrobenzene sulphonic acid-induced colitis mouse model.

AIMS: Superoxide dismutase (SOD) can prevent and cure inflammatory bowel diseases by decreasing the amount of reactive oxygen species. Unfortunately, short half-life of SOD in the gastrointestinal tract limited its application in the intestinal tract. This study aimed to investigate the treatment effects of recombinant SOD Lactobacillus fermentum in a colitis mouse model. METHODS AND RESULTS: In this study, we expressed the sodA gene in Lact. fermentum I5007 to obtain the SOD recombinant strain. Then, we determined the therapeutic effects of this SOD recombinant strain in a trinitrobenzene sulphonic acid (TNBS)-induced colitis mouse model. We found that SOD activity in the recombinant Lact. fermentum was increased by almost eightfold compared with that in the wild type. Additionally, both the wild type and the recombinant Lact. fermentum increased the numbers of lactobacilli in the colon of mice (P < 0·05). Colitis mice treated with recombinant Lact. fermentum showed a higher survival rate and lower disease activity index (P < 0·05). Recombinant Lact. fermentum significantly decreased colonic mucosa histological scoring for infiltration of inflammatory cells, lipid peroxidation, the expression of pro-inflammatory cytokines and myeloperoxidase (P < 0·05) and inhibited NF-κB activity in colitis mice (P < 0·05). CONCLUSIONS: SOD recombinant Lact. fermentum significantly reduced oxidative stress and inflammation through inhibiting NF-κB activation in the TNBS-induced colitis model. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides insights into the anti-inflammatory effects of SOD recombinant Lact. fermentum, indicating the potential therapeutic effects in preventing and curing intestinal bowel diseases.

Protein transduction domain-hA20 fusion protein protects endothelial cells against high glucose-induced injury.

We constructed a plasmid containing a protein transduction domain (PTD) and a human A20 (hA20) gene fragment; the fusion protein was obtained by highly expressing this plasmid in the yeast Pichia pastoris GS115. The plasmid was obtained by adding 9xArg and EcoRІ recognition sites to the end of the primer, and 6xHis-Tag and NotІ recognition sites to its end. After sequencing, the hA20 gene fragment was inserted into plasmid pPIC9k to construct expression vector pPIC9k-PTD-hA20; then, we transfected GS115 with the vector and induced PTD-hA20 protein expression. We purified protein from the yeast fermentation supernatant using a nickel column. Human umbilical vein endothelial cells (HUVECs) were cultured in high glucose medium (30 mM glucose) and in high glucose medium containing different concentrations of protein. Apoptosis of HUVECs was assayed by TUNEL 72 h later. The biological activity tests indicated that the fusion protein not only passed through the cell membrane freely, but also inhibited apoptosis of HUVECs induced by high glucose levels. We conclude that the fusion protein PTD-hA20 has potential for clinical use.

Zinc finger protein A20 overexpression inhibits monocyte homing and protects endothelial cells from injury induced by high glucose.

Diabetes mellitus causes vascular lesions and may ultimately lead to atherosclerosis. One of the earliest steps in the development of atherosclerotic lesions is the adhesion of monocytes to endothelial cells of the vessel wall. It is currently unknown whether zinc finger protein A20 is able to protect endothelial cells from injury caused by high levels of glucose and monocyte homing. In our study, adhesion of monocytes to the vessel wall endothelium was detected by measuring the rolling velocity of monocytes along human umbilical vein endothelial cells (HUVECs). Activation of NF-κB was analyzed through Western blot. HUVEC apoptosis was monitored by TUNEL in situ end-labeling and flow cytometry. High glucose concentrations (25 mM) stimulated monocytes, reducing the velocity at which they roll along HUVECs. Stimulation of monocytes with high levels of glucose also induced HUVEC apoptosis. Overexpression of the zinc finger protein A20 inhibited monocyte recruitment, NF-κB activation, P-selectin expression, and HUVEC apoptosis induced by high glucose levels. We conclude that zinc finger protein A20 can protect HUVECs from injury induced by high levels of glucose and potentially could be used to develop treatments against diabetic vascular lesions.

Optimal timing of operation for repairing atonic bladder after medullary cone injury: an experimental study in rats.

OBJECTIVE: To determine optimal timing of operation for repairing atonic bladder after medullary cone injury in rats. MATERIALS AND METHODS: I n all, 56 adult female Sprague-Dawley rats were equally randomized into seven groups: normal control group, and 4w, 6w, 8w, 10w, 12w and 16w groups after medullary cone injury, assigned as groups A-G. The model was established by sharp transaction of spinal cord at the level of L(4/5) vertebral body. Bladder weight, cross-sectional area and ultrastructure of the detrusor muscle and its neuromuscular junction (NMJ), fibrotic change, and alpha-smooth muscle antibody (alpha-SMA) expression in the detrusor muscle were examined individually. RESULTS: Bladder weight in groups E-G was significantly increased than that in group A (P<0.05). And cross-sectional area of detrusor muscle fiber in groups E-G was significantly smaller than that in group A (P<0.05). Transmission electronic microscopy showed that the number of synaptic vesicles, mitochondria and other organelles in NMJ decreased markedly in group E. In groups F and G, NMJ further degenerated with synaptic vesicles and organelles decreased or even disappeared. Masson's stain showed that the proportion of connective tissue in the detrusor muscle of groups E-G was significantly different from that of group A (P<0.05). alpha-SMA expression in the detrusor muscle decreased with the lapse of time. CONCLUSIONS: The 10th week after rat medullary cone injury can be regarded as the time node when the detrusor muscle and NMJ undergo changes, and therefore surgical nerve repair should be performed before this.

[The diagnosis and treatment of quadrilateral space syndrome].

OBJECTIVE: To investigate the compression feature, clinical manifestation and the results of treatment of quadrilateral space syndrome. METHODS: Four patients with axillary nerve entrapment at quadrilateral space had been treated and followed up for 5 to 12 months from May 1999 to June 2000. The causes, symptoms, signs and the treatment management of those cases were analyzed. RESULTS: Among the 3 cases which received operation, sensation and motor function completely recovered in 2 cases and partially recovered in 1 case. No obvious recovery of sensation and motor function in the case which received local nerve blocking treatment. CONCLUSION: The main diagnostic evidence for axillary nerve entrapment is the deltoid muscle paralysis and paresthesia in the lateral side of shoulder, and early neurolysis is recommended as soon as the diagnosis is clarified.

[Experimental study on the hemostatic properties of collagen sponge].

OBJECTIVE: To validate the hemostatic properties of collagen sponge made in China. METHODS: The experimental model of superficial cut of liver was established in 20 Sprague-Dawley adult rats, which were divided into two groups randomly. Collagen sponge or gelatin sponge was used to cover the cut respectively. Hemostatic result was observed. Afterwards, standard liver trauma model by resection left front liver lobe was made, wound was treated with collagen sponge or gelatin sponge respectively. Hemostatic result was observed. Concurrent hemostatic time and bleeding amount were noted. At 7, 14 and 20 days after operation, intra-abdominal adhension, infection and healing state of liver were observed by exploratory laparotomy. The histological changes of regenerate liver tissue were observed by microscopy. RESULTS: Collagen sponge adhered to wound well. Concurrent hemostatic time and bleeding amount in collagen sponge group were superior to those of gelatin sponge (P < 0.05). The histological examination showed that collagen sponge was absorbed and degraded rapidly, regenerative hepatocytes could be induced. CONCLUSION: Collagen sponge has fine hemostatic properties and can induce regeneration of hepatocytes effectively. It is worth popularizing for its convenience in clinical application and its properties of rapid degradation and absorption.

[Experimental study on combining selective rhizotomy of different anterior and posterior sacral roots for restoration of bladder function after spinal cord injury].

OBJECTIVE: To investigate an alternative procedure for complete denervation of bladder in the supra-cone cord injury to restore the bladder function. METHODS: Sixteen dogs were included in this study after their spinal cords were transected above the cone. They were divided into 6 groups and performed the rhizotomy of L7 to S3 root in different combination respectively. The bladder and urethra pressure change by electrostimulation during operation and cystometrogram change after operation were tested. RESULTS: 1. Electrostimulation study: for bladder innervation, S2 was the most important and S1 was secondary. While for urethra innervation, S1 was more important than S2. When the anterior and posterior roots of S1 and S2 were intact with rhizotomy of posterior roots of L7 and S3, stimulated the common or posterior root of S1 and S2, the change of pressure in bladder and urethra was the same. When the anterior roots of S1 and S2 were resected with rhizotomy of posterior roots of L7 and S3, the pressure in bladder and urethra was significant decreased compared to stimulating the corresponding posterior roots. 2. Cystometrogram (CMG) study: in the complete deafferented group, resecting the posterior roots of L7 to S3, the bladder became flaccid. While resecting the posterior root of S2 and anterior root of S1 or, resecting the posterior root of S1 and anterior root of S2, combining with rhizotomy of posterior roots of L7 and S3, the CMG curve was similar to the complete deafferented group. In the S1 and S2 intact group, the bladder became spastic. CONCLUSION: Combining rhizotomy of anterior and posterior sacral root in different level has the same effects on bladder as complete deafferentation.

Urologic status of 74 spinal cord injury patients from the 1976 Tangshan earthquake, and managed for over 20 years using the Credé maneuver.

OBJECTIVE: To evaluate the urologic safety of long-term Credé maneuver as bladder management in spinal cord injured patients. METHODS: Seventy-four paraplegics were included in this cross-sectional study. They were injured in the Tangshan earthquake in 1976. All patients have large volume (flaccid) bladders and have practiced the Credé maneuver for more than 20 years to expel urine. Current residual urine volume and urologic complications were investigated. RESULTS: 93.2% of patients have residual urine larger than 100 ml and 50% of cases larger than 300 ml. The prevalence of urologic complications is high: pyuria in 82.4%, urinary lithiasis in 31.3%, ureteral dilatation in 59.5%, hydronephrosis in 35.1% and renal damage in 16.2%. Men are more susceptible to upper urinary tract deterioration than women (P<0.05). CONCLUSION: The Credé maneuver is not safe for long-term use in spinal cord injury patients, especially in men.