Objective To establish a quality control method for detecting impurities in chloral hydrate raw materials, improve the quality standards and control limits of raw materials. Methods The determination methods of chloroform and halogenated carboxylic acid in chloral hydrate were established to monitor the change of impurities in chloral hydrate through stability. Results The research and establishment of chloroform and halogenated carboxylic acid methods met the requirements of relevant regulations for analytical methodology verification, which could accurately detect four impurities in raw materials and preparations by one method. Conclusion The study provides technical support for the improvement and optimization of the quality standards of chloral hydrate and preparations. It is very necessary to implement the impurity monitoring in preparation research and production process by the chloral hydrate impurity detection and the stability comparison of this product at high temperature and light, which could largely promote the safety of medication.

OBJECTIVE: To explore the genetic etiology of a child with microcephaly-cortical blind syndrome. METHODS: Clinical data of the child was collected. The child and her parents were subjected to whole exome sequencing (WES). Candidate variants were validated by Sanger sequencing. RESULTS: WES revealed that the child has harbored compound heterozygous variants c.1051C>T and c.609delA of the DIAPH1 gene. CONCLUSION The compound heterozygous variation c.1051C>T (p.R351X) and c.609delA (p.E203Efs*19) of the DIAPH1 gene probably underlay the microcephaly-cortical blindness syndrome in this child.
Blindness/genetics* ; Child ; Female ; Formins/genetics* ; Genetic Testing ; Humans ; Microcephaly/genetics* ; Mutation ; Pedigree ; Exome Sequencing

Traditional right ventricular pacing is non-physiological pacing, can lead to ventricular electrical and mechanical dyssynchmnization, which increases the risk of heart failure and atrial fibrillation. His-Purkinje system pacing is the hot research of physiological pacing method. The high thresholds and difficult implantation of his bundle pacing limits its application and popularization. In recent years, domestic scholars have proposed left bundle branch area pacing. Many studies have confirmed the safety and feasibility of left bundle branch area pacing. Left bundle branch pacing transmit down the conduction system, which can effectively achieve ventricular electrical and mechanical synchrony, At the same time, it also has many others advantages, such as good and stable electrical parameters, significantly improved cardiac function, and pacing across blocked parts. It will become the main development direction of physiological pacing in the future. This article aims to review the origin, standard and classification, advantages and limitations of left bundle branch area pacing.

Objective To investigate the relationships between serum osteocalcin (OC) levels and glycometabolism markers in nondiabetic post-traumatic male patients.Methods Populaitons were selected at the Department of Emergency Medicine of Shanghai Jiao Tong University Affiliated Sixth People's Hospital from October 2017 to February 2019.The age,injury severity score (ISS),and characteristic indicators were recorded.The inclusion criteria were age ≥ 18 years and blood collection time ＜ 24 h after the injury.The exclusion criteria were emergency surgery,acute brain trauma,and hemoglobin A1c (HbA1c) ≥ 6.0％.The patients were divided into two groups by fasting plasma glucose (FPG):stress hyperglycemia (SH) (FPG＞7.8 mmol/L) and nonstress hyperglycemia (NO-SH) (FPG ≤ 7.8 mmol/L) groups.The fasting venous blood samples were collected and examined.The characteristics and biochemical indicators in the two groups were compared statistically by LSD-t test,rank sum test and ANOVA,and the relationships between serum OC levels and glycometabolism markers were analyzed by partial correlation analysis.Results A total of 395 traumatic patients were enrolled and divided into the SH group (n=182) and NO-SH group (n=213).There were no differences in ISS,fasting insulin (FINS),and C-peptide (C-P) levels between groups.Age,HbAlc and FPG were higher (P=0.041,P=0.037,P＜0.01),while the OC level was lower (P=0.023),in the SH group than those in the NO-SH group.The serum OC level did not correlate with HbAlc,FPG,and FINS,but negatively correlated with C-P by partial correlation analysis (r=-0.262,P=0.008).The multivariate linear regression analysis showed that C-P was an independent factor affecting serum OC levels after trauma (β=-0.655,P=0.043).Conclusion A correlation existed between the serum OC level and glycometabolism markers in nondiabetic post-traumatic male patients.

Objective: To investigate the effect of 2, 2', 4, 4'-tetrabromodiphenyl ether (PBDE-47) on the mitochondrial mass in rat adrenal pheochromocytoma (PC12) cells and the potential mechanisms. Methods: Highly differentiated PC12 cells were divided into control, 1, 10 or 20 μmol/L PBDE-47-treated groups and cultured for 24 h. Transmission electron microscopy was employed to observe the changes in mitochondrial morphology and quantity in PC12 cells. Flow cytometry was used to measure the fluorescence intensity of Nonyl Acridine Orange (NAO) , a fluorescent indicator of mitochondrial membrane cardiolipin, to reflect mitochondria mass. Western blotting was used to determine the expression levels of Mitofusion 1 (Mfn1) and Fission 1 (Fis1) proteins. To further explore the role of abnormal mitochondrial fusion and fission in PBDE-47-induced mitochondrial mass changes, PC12 cells were divided into control group, 5 μmol/L M1 treatment group, 20 μmol/L PBDE-47 treatment group and 5 μmol/L M1+20 μmol/L PBDE-47 combined treatment group and cultured for 24 h, then the fluorescence intensity of NAO and expression levels of Mfn1 and Fis1 proteins were detected. Results: The control group showed numerous mitochondria with normal morphology, while the number of mitochondria decreased after PBDE-47 treatment. Especially, the disappeared cristae, swelling and vacuoles of mitochondria and decreased fluorescence intensity of NAO (P<0.05) were observed in 10 and 20 μmol/L PBDE-47-treated groups. Meanwhile, the expression levels of Mfn1 and Fis1 proteins in the 10 and 20 μmol/L PBDE-47-treated groups were significantly decreased compared with control group (P<0.05) . However, 5 μmol/L M1 co-treatment with 20 μmol/L PBDE-47 significantly increased the levels of Mfn1 and Fis1 proteins and fluorescence intensity of NAO compared with the 20 μmol/L PBDE-47 group (P<0.05) . Conclusion PBDE-47 can inhibit the mitochondrial fusion and fission process, thus leading to damage of mitochondria mass in PC12 cells.

OBJECTIVE:To establish the method for simultaneous determination of berberine hydrochloride and baicalin in Jianpi zhixiening granules. METHODS:HPLC switching walvelength method was adopted. The determination was performed on Hypersil BDS C18 column with mobile phase consisted of methanol-0.45％ phosphoric acid solution-triethylamine(50:49:1,V/V/V) at the flow rate of 1.0 mL/min. The detection wavelength was set at 265 nm(berberine hydrochloride)and 280 nm(baicalin). The column temperature was set at 30 ℃,and sample size was 10 μL. RESULTS:The linear range of berberine hydrochloride and baicalin were 60.3-312.8 ng(r=0.9997)and 81.5-368.9 ng(r=0.9999). The limits of quantitation were 0.6668,0.7740 ng,andthe limits of detection were 0.2226,0.2580 ng,respectively. RSDs of intermediate precision,stability and repeatability tests were all lower than 1.0％. The recoveries were 96.48％-99.30％(RSD=1.06％,n=6) and 95.20％-99.39％(RSD=1.66％,n=6), respectively. RSDs of durability test were all lower than 2.0％. CONCLUSIONS:The method is simple, precise, stable, reproducible,accurate and durable. It can be used for simultaneous determination of berberine hydrochloride and baicalin in Jianpi zhixiening granules.

OBJECTIVE:To establish the method for simultaneous determination of the content of berberine hydrochloride, baicalin and osthole in Jinchan zhiyang capsules. METHODS:HPLC method was adopted. The determination was performed in Hypersil BDS C18 column with mobile phase consisted of methanol-acetonitrile-0.1%phosphoric acid-three triethylamine(50∶30∶19∶1, V/V/V/V) at the flow rate of 1.0 mL/min. The detection wavelengths were set at 265 nm (berberine hydrochloride),280 nm (baicalin)and 322 nm(osthole). The column temperature was set at 30 ℃,and sample size was 10 μL. RESULTS:The linear range of berberine hydrochloride,baicalin and osthole were 80-800 ng(r＝0.999 8),60-600 ng(r＝0.999 9),60-600 ng(r＝0.999 6),respectively. RSDs of precision,stability and reproducibility tests were all lower than 2.0%. The limits of quantitation were 80,60,60 ng,respectively,and the limits of detection were 24,20,20 ng,respectively. The recovery rates were 97.4%-98.3%(RSD＝0.33%,n＝6),98.4%-99.6%(RSD＝0.42%,n＝6)and 96.9%-99.0%(RSD＝0.92%,n＝6),respectively. RSDs of durability tests were all lower than 1.2%. CONCLUSIONS:The method is simple, accurate, precise, stable, reproducible and durable. It can be used for simultaneous determination of berberine hydrochloride,baicalin and osthole in Jinchan zhiyang capsules.

Abstract BACKGROUND:Caspase-8 plays an important role in starting the process of cel apoptosis, and diacetylmorphine can induce neuronal apoptosis. The relationship between the apoptosis of cerebelar granule neurons cels induced by diacetylmorphine and caspase-8 has not been reported. OBJECTIVE:To investigate the effect of caspase-8 on diacetylmorphine-induced neuronal apoptosis. METHODS:Cerebelar granule neurons from Sprague-Dawley rats aged 7 days were culturedin vitro. At 7 days, the cels were cultured with different dosage of diacetylmorphine (10, 40, 80, 100, 120 mg/L) and SP600125 for 24 hours, and were divided into control group, 80 mg/L diacetylmorphine group, diacetylmorphine+SP600125 group. Cel morphology was observed by Hoechst 33258 fluorescent staining, and cel apoptosis rate was measured by flow cytometry. Immunofluorescence staining, RT-PCR and western blot assay were used to detect caspase-8 mRNA and protein expression. RESULTS AND CONCLUSION:(1) After different dosage of diacetylmorphine was used to induce neuronal apoptosis, dark blue translucent apoptotic bodies were found in apoptotic cels, appearing with nucleus condensation, cohesion and fracture, and the apoptosis rate presented an increasing trend with increasing of drug dosage (P < 0.05). (2) Compared with the control group, caspase-8 mRNA and protein expression was remarkable under the intervention of 80 mg/L diacetylmorphine(P < 0.05); caspase-8 mRNA expression exhibited an increasing trend with increasing dosage of diacetylmorphine (P < 0.05), while caspase-8 mRNA and protein expression in the diacetylmorphine+SP600125 group was significantly lower than that in the 80 mg/L diacetylmorphine group (P < 0.05). These findings indicate that caspase-8 is involved in the process of diacetylmorphine-induced neuronal apoptosis, and meanwhile, it is also an important candidate of pro-apoptotic factors in the c-jun N-terminal kinase signaling pathway.

BACKGROUND:Calcium channel abnormalities can damage myocardium. Heroin can directly affect calcium ion channel, and alter myocardial structure.
OBJECTIVE:To study the changes of heroin-induced myocardial ultrastructure, L-type calcium channel current and action potential of myocardial cel s after rat cardiac arrhythmia.
METHODS:Sprague-Dawley rats were randomly divided into control group and model group. In the model group, rats were administered heroin at initial dose of 5 mg/kg?d. The daily dose escalation method was used (increasing dose:2.5 mg/kg?d) to replicate rat models of heroin addiction. At 20 days, models of heroin addiction were successful y established. At 30 days after increasing the dose, rat models of heroin addiction-induced arrhythmias were further established.
RESULTS AND CONCLUSION:Compared with the control group, the electron microscopy demonstrated that myocardial structure changes mainly presented nuclear membrane shrinkage, nuclear condensation, nuclei became smal , chromatin assembled into blocks, mitochondria disordered and disappeared, sarcomeres disordered, focal fracture, and unclear myofilament in rat models of heroin addiction. Electric current-voltage curve of myocardial cel L-type calcium channel current showed the increasing trends. The 90%repolarization action potential was significantly shortened. These data indicated that heroin can directly lead to the pathological change of myocardial structure. Calcium channel current change is one of the main reasons for myocardial injury.

Objective To explore the value of fluorescence in situ hybridization (FISH) and multiplex RT-PCR in the detection of mixed lineage leukemia (MLL) gene rearrangement in acute leukemia (AL) patients. Methods Dual-color MLL probe, multiplex RT-PCR and R or G banding techniques were used to detect the MLL gene rearrangement in 189 cases of AL.Results MLL gene rearrangements were detected in 9 cases (5.03 ％) by FISH,and 16 cases (8.47 ％) by multiplex RT-PCR,including MLL/AF9,MLL/AF10,MLL/AF6, MLL/AF7, MLL/ELL, MLL/PTD. R or G banding techniques could find 11q23 in 5 out of 189 patients (2.65 ％). There was no statistic difference in the incidence of 6 common MLL gene rearrangements between ALL (73 cases) and AML patients (116 cases) (P ＞ 0.05).Conclusion Multiplex RT-PCR is a powerful technique in the detection of MLL gene rearrangement for tentatively diagnosed AL.It could not only confirm translocation detected by conventional cytogenetic method, but also detect MLL partial tandem duplication which could not been detected by cytogenetic examination or FISH. It plays an important role in guiding therapy and predicting prognosis for AL.