The rapid changes in the research and development environment of new anti-tumor drugs in China have brought various challenges to drug innovation. How to explore the clinical advantages of new drugs in the early phase, and design scientific, reasonable and efficient pivotal clinical trials for drug registration accordingly, is one of the key challenges. This article takes innovative new drugs for hematological malignancies as an example, comprehensively elaborates the considerations on the timing for entering the pivotal clinical trial and the key elements of the trial design from the perspective of clinical reviewers.

Hemophilia is the only rare hereditary hemorrhagic disorder included in the First Rare Diseases catalogue. However, rare bleeding diseases identified in the clinic are far more common than hemophilia. Most other rare hemorrhagic disorders have less effective treatment than hemophilia. Hemophilia has a history of successful drug development in rare hemorrhagic diseases, and the cycle between clinical research and drug development has been gradually realized. Drug research and pharmaceutical companies can refer to the drug research and development process in the field of hemophilia, learn from the experience of hemophilia drug research and develop treatments. The industry can increase drug development by strengthening basic research, focusing on the value of natural history research, the application of quantitative pharmacological tools and improving the efficiency of drug development to meet the urgent unmet medical needs of patients with rare hemorrhagic diseases.

The clinical data of 69 patients with non-HIV-related cryptococcal meningitis admitted in the Second Hospital of Hebei Medical University from January 2013 to May 2019 were analyzed retrospectively.The main presentations of 69 patients are headache,fever,nausea,vomiting, visual impairment, hearing damage.Among them, 36 cases (52%) had underlying diseases, 30 cases (43%) were misdiagnosed, and 38 cases (55%) were complicated with high intracranial pressure. Cerebrospinal fluid examination showed that leukocytes increased in 47 cases, protein increased in 55 cases, chloride decreased in 41 cases, glucose decreased in 34 cases. The imaging findings were cerebral ischemia, hydrocephalus, meningeal or cerebral parenchyma enhancement. During the induction period, 63 cases were treated with combined antifungal drugs and 6 cases were treated with single antifungal drugs. The clinical symptoms were improved in 54 cases, 9 cases were discharged automatically and 6 cases died. The clinical manifestations, routine and biochemical examination of cerebrospinal fluid, imaging findings are not specific for patients with non-HIV-related cryptococcal meningitis.So early and multiple lumbar puncture should be performed to find etiological evidence to reduce misdiagnosis. The combination of antifungal drugs during the induction period is safe and effective.

Objective To investigate the relationship between rs17525495 locus polymorphism of leukotriene A4 hydrolase (LTA4H) gene and the severity of tuberculous meningitis (TBM). Methods A total of 184 TBM patients from Department of Neurology, the Second Hospital of Hebei Medical University from January 2014 to October 2016 were selected as research subjects. According to the British Medical Research Council criteria, the severity of TBM patients was divided into three stages. The single nucleotide polymorphism rs17525495 of LTA4H gene was sequenced, and the general case data, clinical manifestations and results of lumbar puncture were analyzed. Results There were 91 cases (49.5％) of CC genotypes of rs17525495 locus in LTA4H gene of 184 cases, 75 cases (40.8％) of CT genotypes and 18 cases (9.8％) of TT genotypes. The frequency of allele C was 69.8％ and T was 30.2％. Patients with different genotypes were compared for their severity, clinical manifestations and lumbar puncture results. Among CC patients, the proportion of stage Ⅰ patients(54.9％, 50/91)was higher than that of stage Ⅱ(22.0％, 20/91)and Ⅲ(23.1％, 21/91). Among TT patients, the proportion of patients with stage Ⅱ(8/18)and Ⅲ(8/18)was higher than patients with stageⅠ(2/18)(χ2=15.898,P=0.003). The incidence of headache, fever, nausea and vomiting, neck stiffness, epilepsy and disturbance of consciousness was statistically analyzed. Compared with CC and CT patients, the incidence of fever(TT:13/18,CC:42/91,CT:50/75,χ2=8.932,P=0.011)and neck stiffness(TT:12/18,CC:38/91,CT:46/75,χ2=7.993,P=0.018)was higher in TT patients. Headache, nausea and vomiting, disturbance of consciousness, and the incidence of epilepsy showed no statistically significant difference. And there was no statistically significant difference in lumbar puncture pressure, chloride, protein and glucose between different genotypes. Conclusion TBM patients with mild illness frequently prompt LTA4H gene rs17525495 locus for the CC type;while patients with severe disease prompt TT type.

Objective To investigate the features of the cerebrospinal fluid (CSF) in the modified ZeiM-Neelsen (MZN) positive tuberculous mengningitis (TBM).Methods We retrospectively reviewed the clinical data of 170 patients with tuberculous meningitis confirmed by MZN stain from December 2012 to July 2015.The purpose of the present study was to investigate the relationship of MZN staining and CSF cytology.Results Among 170 patients with TBM confirmed by MZN staining,128 cases had first detectable acid-fast bacillus (AFB) in earlier stage.The cytology included 15.5％ mixed cellular cytology,58.5％ lymphoid cytology,19.5％ neutrophilic cytology and 6.5％ normal cytology.Twenty-four cases had first detectable AFB within 1-2 months following disease onset.The cytology included 13.1％ mixed cellular cytology,56.6％ lymphoid cytology,21.7％ neutrophilic cytology and 8.7％ normal cytology.Eighteen cases had first detectable AFB 2 months after disease onset.The cytology included 26.7％ mixed cellular cytology,46.7％ lymphoid cytology,20.0％ neutrophilic cytology,6.6％ normal cytology.There was no significant difference in median time of first detectable AFB among those four types of cytology (P=0.812).There was significant difference in median time of first detectable AFB between patients with and without anti-TB therapy [21.5 (12.3,37.8) days vs.8.5 (6.0,16.3)days,P＜0.001].There was no significant difference in median time MZN stain turning negative between patients with and without anti-TB therapy [11 (5.75,19.25) days vs.6(4.25,10.75)days,P=0.230].Conclusions AFB can be detectable within a month after the onset of TBM in most of cases.(MZN) positive staining is not associated with the major type of cytology.Anti-TB therapy may delay the first detectable time of AFB.

Objective To investigate the role of S100B protein in the pathogenesis of patients with Japanese encephalitis (JE).Methods A total of 45 patients were enrolled in the Second Hospital of Hebei Medical University from August 2013 to October 2013,who were diagnosed as JE on the basis of clinical features and positive IgM antibodies against JE virus measured by enzyme-linked immunosorbent assay (ELISA) from the Center of Disease Control of Shijiazhuang.The JE patients were divided into initial phase group,acute phase group and convalescence group based on the course,mild JE group,moderate JE group and severe JE group based on the severity,MRI-no-lesion group and MRI lesions group based on the imaging findings of JE.Twelve cases with no evidence of infection in central nervous system in the meantime were chosen as control.The S100B protein was measured by ELISA.Results The content of S100B protein in cerebrospinal fluid was as follows:522.76 (393.35,620.37) pg/ml in mild JE group (acute phase group:609.77 (549.27,779.71) pg/ml,convalescence group:420.48 (344.36,453.19) pg/ml),792.09 (705.47,1 108.96) pg/ml in moderate JE group (acute phase group:770.19 (646.31,1 069.54) pg/ml,convalescence group:803.45 (602.90,1 396.84) pg/ml),and 1 021.94 (680.84,1 302.15) pg/ml in severe JE group (acute phase group:981.82 (680.84,1 826.28) pg/ml,convalescence group:989.00 (553.62,1 207.67) pg/ml).The S100B protein content was 561.52 (454.36,814.56) pg/ml,803.45 (602.90,1 104.01) pg/ml,762.22 (594.95,1 044.97) pg/ml,581.76 (442.51,1 069.10) pg/ml in MRI-no-lesion group,MRI lesions group,total acute phase group and total convalescence group,respectively.While in control group,the S100B protein content was 266.71 (205.72,390.05) pg/ml.The contents of S100B protein in moderate JE group,severe JE group,total acute phase group,total convalescence group,MRI-no-lesion group,MRI lesions group were higher than that in control group (H =4.864,5.497,5.075,3.918,2.971,4.981,P =0.000,0.000,0.000,0.000,0.009,0.000).The contents of S100B protein in mild JE group was lower than that in moderate JE group and severe JE group (H =-2.786,-3.514,P =0.032,0.003).Conclusions The level of S100B protein in cerebrospinal fluid is related with the severity,duration and imaging presentation of JE patients.The dynamic monitoring of S100B protein levels is of great significance for assessment of the patients' condition and curative effect.

Objective To explore the diagnostic significance of Xpert MTB/RIF in cerebrospinal fluid,and evaluate the application for early diagnosis of tuberculous meningitis(TBM).Methods Sixty cases of TBM and 30 cases of non-TBM patients were selected as our subjects.Xpert MTB/RIF and modified Ziehl-Neelsen stain were performed in cerebrospinal fluid.The detection rate of the system and the resistance of the patients were analyzed.Results Eleven cases were diagnosed as the positive cases in 60 cases with TBM,and 0 case was diagnosed as TBM in control group.Sensitivity and specificity of Xpert with TBM were 18.33％ and 100％,respectively.The difference of the two groups was statistically significant (P =0.014).The positive rate of definite group was 23.68％(9 cases),18.18％(2 cases) in probable group and 0％ in possible group,and the difference of the three groups was statistically significant(x2 =3.070,P＞0.05).The resistance rate was 36.36％ (4/11).Sensitivity of the modified Ziehl-Neelsen staín was 63.33％ (38/60).Eleven cases were detected positive by Xpert MTB/RIF,9 cases were positive with modified acid fast staining,and the positive rate was 18.33％,and the difference of the two methods was statistically significant (P =0.000).Conclusion Xpert MTB/RIF test is simple and rapid diagnostic method.The combination of Xpert MTB/RIF and modified ZiehlNeelsen stain will improve the efficiency of the early diagnosis of TBM.

Objective Toevaluatea modified Ziehl-Neelsen(Z-N) stain in the diagnosis of tuberculous meningitis. Methods Cerebrospinal fluid specimens from 35 patients were stained by using the modified Ziehl-Neelsen staining. Re-sults The positive rate was 94.29% in 35 patients with tuberculous meningitisand the intracellular acid-fast bacilli was detected in 53.40%of all specimens. One case was stained positive in 15 patients with non-tuberculous meningitis. Con-clusion The modified Ziehl-Neelsen stain not only significantly improves the detection rates of tuberculous meningitisbut alsois able to identify intracellular M.tuberculosisin cerebrospinal fluidspecimen.Thus, the modified Z-N stain can be a convenient tool for diagnosing tuberculous meningitis.

Objective To investigate the effect of free transplantation of fibula composite tissue flap to the repair of forearm bone and soft tissue serious defect.Methods Eleven cases of reparing forearm bone and soft tissue serious defect through transplantation of free fibula composite tissue petal were applied from March 2004 to February 2011.The length of transplanted fibula composite tissue flap was 8-14 cm ; the flap area was 5 cm ×8 cm-20 cm ×20 cm.The curing situation on bone fracture was observed in 3,6 and 12 months after the surgical opration and the function of defected arm was evaluated in 1 year after surgical operation.Results All of 11 cases of fibula composite tissue flap were survived.The observation was undertaken for more than 12 months after the operation and the fracture section occured the characteristics of healing up in 3 months and fibula and arm bone occured well healed up in half a year; It scored 22.9 according to Enneking system after 1 year of the operation.The function of forearm rotation were classed as this:3 good cases,6 medium cases and 2 poor cases.In the 2 sural nerve bridging transplantation cases,one case was repaired of radial nerve inside static's two-point discrimination (s2PD) to 9 mm,another case was repaired of ulnar nerve distal volar little finger s2PD to 15 mm.All the cases could achieve making a fist with thumb and a thumb could be oppoiste to other 4 fingers,and the ankle joint movement was normal.Conclusion Transplantion of free fibula composite tissue flap to the repair of forearm bone and soft tissue serious defect is an ideal surgical operation method.

ObjectiveTo investigate the significance of cerebrospinal fluid(CSF) S-100B protein and vascular endothelial growth factor (VEGF) levels in the pathogenesis of brain injury of viral encephalitis.MethodsForty-two patients with viral encephalitis (viral encephalitis group) and 40 patients with other disease at the corresponding time period(control group) were involved in this study.CSF (routine,biochemistry and cytology) was detected,and the levels of S-100B protein and VEGF in CSF were detected by ABC-ELISA method.ResultsWhite blood cell count was (0-584) × 106/L in viral encephalitis group,and (0-200) × 106/L in control group.The levels of protein and glucose in CSF had no significant difference between two groups (P＞ 0.05),and the level of chloride in CSF in viral encephalitis group was significantly lower than that in control group[ ( 110.10 ± 31.22 ) mmol/L vs.( 123.80 ± 6.32 ) mmol/L ] (P =0.006).In viral encephalitis group,cytological examination showed that mixed type cytological reaction was in 6 cases (14.3％,6/42).The level of S-100B protein in viral encephalitis group [25.04-47.97 (28.37 ± 6.09) ng/L] was significantly higher than that in control group[ 25.04-29.64(26.03 ± 0.90) ng/L ] (t =2.462,P =0.018).The level of VEGF in viral encephalitis group[88.84～143.77(96.24 ± 13.38) ng/L] was significantly higher than that in control group [89.15～96.18 (90.67 ± 1.71 ) ng/L] (t =2.673,P =0.011 ).ConclusionsThe high levels of S-100B protein and VEGF in CSF could support the viral encephalitis diagnosis.Tracking the levels of S-100B protein and VEGF in CSF dynamically have noticeable effect on checking the condition of viral encephalitis patients.