OBJECTIVE:To establish a method for the determination of related substances in bifonazole raw material. METH-ODS:HPLC method was performed on the column of Kromasil C18 with mobile phase of methanol-0.02 mol/L phosphoric acid(ad-justed pH to 7.5 with triethylamine)(70:30,V/V)at a flow rate of 1.0 ml/min,detection wavelength was 258 nm,volume injection was 20 μl,and the column temperature was 40 ℃. RESULTS:The linear range was 0.05-0.25 μg/ml for bifonazole(r=0.9996), 0.05-0.25 μg/ml for impurity A(bifonol)(r=0.9997)and 0.05-0.25 μg/ml for impurity B(4-C isomer)(r=0.9995);the detec-tion limits were 8.2 ng/ml,7.5 ng/ml and 8.4 ng/ml,and the quantification limits were 27.1 ng/ml,24.7 ng/ml and 27.8 ng/ml,re-spectively;RSDs of precision and reproducibility tests were lower than 1%;recovery of impurity B was 95.13%-101.29%(RSD=1.89%,n=9);both impurity A and impurity B were were detected in the 3 batches of samples. CONCLUSIONS:The method is accurate,sensitive and reproducible,and can be used for the determination of the related substances in bifonazole raw material.

BACKGROUND: Although there are many studies on acute patellar dislocation in adolescents, the optimal treatment strategy is under discussion.OBJECTIVE: To summarize the clinical efficacy of reconstruction of the medial patellofemoral ligament (MPFL) in the treatment of adolescent acute patellar dislocation using double-pulley technique.METHODS: Thirty cases of acute patellar dislocation were enrolled and received reconstruction of the MPFL using double-pulley technique. We proceeded to prepare the bone bed for anchor placement at the medial point of the superior edge of the patella and at the upper 1/3 patella along the long axis of the patella using arthroscopic grinding. Two double-loaded 5.0 anchors were respectively placed at the medial point of patella and at the upper 1/3 patella parallel to the patellar articular surface, and then tied using the double-pulley technique. Long longitudinal incisions were performed at the medial condyle and at the adductor tubercle to expose these two structures, the deep fascias tunnel between two incisions were penetrated to pull the tendon weave end through the tunnel until the lateral side along a femoral tunnel; finally the tendon was fixed using screws in appropriate tightness. All patients were followed for more than 10 months, and the patellar-related parameters and knee function parameters were compared before and after operation. RESULTS AND CONCLUSION: There were 12 cases of intra-articular cartilage injury and 4 cases of meniscus injury. The patella title angle, Q-angle and outward shift distance at the last follow-up were less than those before operation. Lysholm and International Knee Documentation Committee scores at the last follow-up were higher than those before operation. The mean operation time was (90±10.5) minutes. There was one patient with knee movement angle from 0° to 30° and one patient with a patellar fracture caused by slipping and treated with internal fixation. None of patients appeared with patellar dislocation, positive apprehension test infection or poor wound healing. These results indicate that the simple and mini-invasive double-pulley technique is a good treatment strategy for acute patellar dislocation, because it is consistent with patellar anatomical stability, holds patellar trajectory visibility, quick recovery and good curative effect. Additionally, osteotomy is recommended when the simple efficacy is not satisfactory.

Objective To investigate the association between chromosome variations,abnormalities and male reproductive hor-mones level with spermatogenesis.Methods The chromosome karyotype,serum reproductive hormone including FSH,LH,T,PRL and E2,and semen were detected in 147 patients with male infertility or recurrent sponotaneous abortion.The results were per-formed the comparative analysis.Results Serum FSH,LH level and the incidence rate of azoospermia in the chromosome abnormal-ity group were significantly higher than those in the chromosome variation group and the normal group(P <0.05,P <0.01),serum T level was significantly lower than that in the chromosome variation group and the normal group(P <0.05).Serum FSH and the incidence rate of oligospermia in the Y chromosome variation group were significantly higher than those in the autosomal variation group(P <0.05).The incidence rate of azoospermia had no statistically significant difference between the Y chromosome variation and the autosomal variation group(P >0.05).Serum FSH,LH level and the incidence rate of azoospermia in the sex chromosome abnormality group were obviously higher than those in the autosomal abnormality group(P <0.05),the serum T level was signifi-cantly lower than that in the autosomal abnormality group(P <0.05).Conclusion The chromosome variation and abnormality are closely related with the reproductive hormones disorder and spermatogenetic function disorder.The obvious increase of serum FSH, LH level and obvious decrease of T level caused by sex chromosome variation and abnormality is one of the pathogenesis of oligo-spermia and azoospermia.

Na⁺/H⁺ exchangers (NHEs) have been shown to be involved in regulating cell volume and maintaining fluid and electrolyte homeostasis. Pooled evidences have suggested that loss of Na⁺/H⁺ exchanger isoform 8 (NHE8) impairs intestinal mucosa. Whether NHE8 participates in the pathology of infectious colitis is still unknown. Our previous study demonstrated that somatostatin (SST) could stimulate the expression of intestinal NHE8 so as to facilitate Na⁺ absorption under normal condition. This study further explored whether NHE8 participates in the pathological processes of infectious colitis and the effects of SST on intestinal NHE8 expression in the setting of infectious colitis. Our data showed that NHE8 expression was reduced in Citrobacter rodentium (CR) infected mice. Up-regulation of NHE8 improved diarrhea symptom and mucosal damage induced by CR. In vitro, a similar observation was also seen in Enteropathogenic E. coli (EPEC) infected Caco-2 cells. Seglitide, a SST receptor (SSTR) 2 agonist, partly reversed the inhibiting action of EPEC on NHE8 expression, but SSTR5 agonist (L-817,818) had no effect on the expression of NHE8. Moreover, SST blocked the phosphorylation of p38 in EPEC-infected Caco-2 cells. Taken together, these results suggest that enhancement of intestinal NHE8 expression by SST could ameliorate the symptoms of mice with infectious colitis.
Absorption ; Animals ; Anti-Inflammatory Agents ; Caco-2 Cells ; Cell Size ; Citrobacter rodentium ; Colitis* ; Diarrhea* ; Enteropathogenic Escherichia coli ; Homeostasis ; Humans ; In Vitro Techniques ; Intestinal Mucosa ; Mice* ; Pathologic Processes ; Pathology ; Phosphorylation ; Somatostatin* ; Up-Regulation*

ObjectiveTo explore the intervention effect and mechanism of Buzhong Yiqiwan （BZYQ） on colitis mice. MethodSixty-four C57BL/6 mice were randomly divided into 2 weeks blank group， 2 weeks model group， 2 weeks high-dose BZYQ （12 g·kg^-1） group， 2 weeks low-dose BZYQ （6 g·kg^-1） group， 4 weeks blank group， 4 weeks model group， 4 weeks high-dose BZYQ （12 g·kg^-1） group， and 4 weeks low-dose BZYQ （6 g·kg^-1） group. The colitis model was induced in mice by feeding 3% dextran sodium sulfate （DSS） for 7 days. The mice received BZYQ （12 and 6 g·kg^-1） by gavage on the 8^th day after modeling， once per day， and sacrificed on the 2^nd and 4^th weeks， correspondingly. The colon length and weight of mice in each group were measured. Hematoxylin-eosin （HE） staining was used for pathological observation and colonic mucosal inflammation was scored. The mRNA and protein expression of NOD-like receptor thermoprotein domain 3 （NLRP3）， apoptosis-associated speck-like protein containing a CARD （ASC）， and cysteinyl aspartate-specific protease-1 （Caspase-1） was detected by real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. Enzyme-linked immunosorbent assay （ELISA） was used to detect the content of inflammatory cytokines， such as interleukin （IL）-1β， IL-18， and IL-33 in colonic tissues. ResultCompared with the 2 weeks blank group， the 2 weeks model group showed shortened colon length， increased colon weight （P<0.05）， loss of epithelial cells， destruction of gland structure， infiltration of a large number of inflammatory cells in mucosa and submucosa， local crypt abscess， and increase in mucosal inflammation score （P<0.01） as revealed by light microscopy， elevated levels of IL-1β， IL-18， and IL-33 in colonic tissues （P<0.05）， and increased mRNA and protein expression of NLRP3， ASC， and Caspase-1 （P<0.05）. The intervention of BZYQ （12 g·kg^-1） restored colon length， alleviated mucosal injury （P<0.05）， down-regulated the content of IL-18 （P<0.05）， reduced the mRNA expression of NLRP3 and ASC as well as the protein expression of ASC and Caspase-1 compared with the conditions in the 2 weeks model group. Compared with the 4 weeks blank group， the 4 weeks model group showed decreased colon length， increased colon weight （P<0.05）， decreased glands in the mucosal layer， expansion of glandular cavity， atrophy of crypt， local connective tissue hyperplasia and lymphocyte infiltration， increased inflammation score （P<0.01） as revealed by the light microscopy， increased expression of IL-1β， IL-18， and IL-33 （P<0.05）， and elevated mRNA and expression of NLRP3 inflammasome complex （P<0.05）. Compared with the conditions in the 4 weeks model group， the intervention of BZYQ （12 and 6 g·kg^-1） could improve colon length and weight （P<0.05）， and the intervention of BZYQ （12 g·kg^-1） could also improve the inflammation score of the colon （P<0.05）. Different from the acute stage， the intervention of BZYQ （12 and 6 g·kg^-1） increased the content of IL-33 in the intestinal mucosa and up-regulated the mRNA and protein expression of NLRP3 inflammasome complexes ASC and Caspase-1 （P<0.05）. ConclusionBZYQ can relieve the injury of colitis induced by DSS in mice. The mechanism is related to the regulation of intestinal immune response mediated by NLRP3 inflammasome， and it has different regulatory effects in acute and chronic inflammation stages.